Prostate Cancer Stem Cells: Viewing Signaling Cascades at a Finer Resolution
Xiukun Lin • Ammad Ahmad Farooqi • Muhammad Zahid Qureshi • Mirna Azalea Romero • Sobia Tabassum • Muhammad Ismail
Abstract It is becoming characteristically more understandable that within tumor cells, there lies a subpopulation of tumor cells with ‘‘stem cell’’ like properties and remarkable ability of self-renewal. Many features of these self-renewing cells are comparable with normal stem cells and are termed as ‘‘cancer stem cells’’. Accumulating experimentally verified data has started to scratch the surface of spatio-temporally dysregulated intracellular signaling cascades in the biology of prostate cancer stem cells. We partition this multicomponent review into how different signaling cascades operate in cancer stem cells and how bioactive ingredients isolated from natural sources may modulate signaling network.
Keywords Prostate cancer stem cells Apoptosis Molecular therapeutics Intracellular signaling
Vitamin D and Its Relevance in the Etiopathogenesis of Oral Cavity Diseases
Zuzannna S´ lebioda • Elżbieta Szponar • Barbara Dorocka-Bobkowska
Abstract Vitamin D belongs to a group of fat-soluble secosteroids which assume many roles in the human organism. In humans the most important forms are vitamin D3 and vitamin D2. Their primary function is the regulation of the calcium and phosphorus balance, which promote the growth of healthy bony tissue. Studies over the past few years have revealed a much wider role of vitamin D involving the aging processes, carcinogenesis, the carbohydrate balance as well as the effects on the course of various infections. In this paper we discuss the basic functions of vitamin D in the human body and the mechanisms of its activity and we summarize recent reports on the impact of vitamin D on the oral cavity with a special emphasis on autoimmunologic diseases, including: recurrent aphthous stomatitis, Behc¸et syndrome and Sjo¨gren syndrome.
Keywords Vitamin D Oral cavity diseases Oral mucosa Autoimmunologic conditions
Techniques of Human Embryonic Stem Cell and Induced
Pluripotent Stem Cell Derivation
Jarosław Lewandowski1 • Maciej Kurpisz1
Received: 24 February 2015 / Accepted: 17 November 2015 / Published online: 3 March 2016
Ó The Author(s) 2016. This article is published with open access at Springerlink.com
Abstract Developing procedures for the derivation of
human pluripotent stem cells (PSCs) gave rise to novel
pathways into regenerative medicine research. For many
years, stem cells have attracted attention as a potentially
unlimited cell source for cellular therapy in neurodegen-
erative disorders, cardiovascular diseases, and spinal cord
injuries, for example. In these studies, adult stem cells were
insufficient; therefore, many attempts were made to obtain
PSCs by other means. This review discusses key issues
concerning the techniques of pluripotent cell acquisition.
Technical and ethical issues hindered the medical use of
somatic cell nuclear transfer and embryonic stem cells.
Therefore, induced PSCs (iPSCs) emerged as a powerful
technique with great potential for clinical applications,
patient-specific disease modelling and pharmaceutical
studies. The replacement of viral vectors or the adminis-
tration of analogous proteins or chemical compounds
during cell reprogramming are modifications designed to
reduce tumorigenesis risk and to augment the procedure
efficiency. Intensified analysis of new PSC lines revealed
other barriers to overcome, such as epigenetic memory,
disparity between human and mouse pluripotency, and
variable response to differentiation of some iPSC lines.
Thus, multidimensional verification must be conducted to
fulfil strict clinical-grade requirements. Nevertheless, the
first clinical trials in patients with spinal cord injury and
macular dystrophy were recently carried out with differ-
entiated iPSCs, encouraging alternative strategies for
potential autologous cellular therapies.
Keywords Stem cells Cardiomyocytes Pluri potency
Directed differentiation
Pituitary Microsomal Autoantibodies in Patients with Childhood-
Onset Combined Pituitary Hormone Deficiency: an Antigen
Identification Attempt
Katarzyna Ziemnicka1 • Paweł Gut1 • Monika Goła˛b1 • Grzegorz Dworacki2 •
El_zbieta Wrotkowska1 • Marek Stajgis3 • Katarzyna Katulska3 • Barbara Rabska-
Pietrzak4 • Monika Obara-Moszyn´ ska4 • Marek Niedziela4 • Bartłomiej Budny1 •
Małgorzata Kału_zna1 • Ryszard Was´ko1 • Marek Ruchała1
Received: 15 July 2015 / Accepted: 15 February 2016 / Published online: 12 March 2016
Ó The Author(s) 2016. This article is published with open access at Springerlink.com
Abstract The role of autoimmunization in the patho-
genesis of pituitary disorders is poorly understood. The
presence of pituitary autoantibodies (APA) has been
detected in various pituitary disorders. Their role, however,
remains elusive. Childhood-onset combined pituitary hor-
mone deficiency (CPHD) may be caused by environmental
or genetic factors. In some of patients, causes of the disease
remain unclear and contributions of autoimmune processes
have been postulated. The aim of this study was to identify
the microsomes-derived pituitary antigens (MPA) as
potential immunogenic autoantigens in patients with
hypopituitarism, therefore 62 CPHD patients, 100 healthy
controls and five autoimmune polyglandular syndrome type
II (APS II) patients were included in the study. The clinical
evaluation included hormonal tests and magnetic resonance
imaging of the pituitary. The sources of MPA were pitu-
itary glands taken from autopsies. Isolated MPA were then
separated on SDS-PAGE gel and incubated with sera
obtained from patients and controls. Microsomal APA
were detected using Western blot and radioimmunological
method. In all CPHD and APS II patients and in 9 %
individuals from control group marked immunoreactivity
was detected against MPA. Antibodies showed high
affinity to 67, 60, 50 and 36 kDa MPAs. Since the identi-
fied autoantigens were of unknown nature, an in silico
exploration of UniProt database was applied and indicated
their possible relationship with chaperones, golgins and
already known autoantigens like GAD67. Reactivity
against MPA indicates that these proteins certainly play a
role in the processes undergoing within pituitary of CPHD
patients. The identification and further detailed studies on
their role in the pathogenesis of CPHD should be
continued.
Keywords Anti-microsomal antibodies
Combined pituitary hormone deficiency Immunoblotting
Radioimmunology
Fecal Microbiota Transplantation Inhibits Multidrug-Resistant
Gut Pathogens: Preliminary Report Performed
in an Immunocompromised Host
Jarosław Bilin´ ski1 • Paweł Grzesiowski2 • Jacek Muszyn´ ski3 • Marta Wro´blewska4,5 •
Krzysztof Ma˛dry1 • Katarzyna Robak1 • Tomasz Dziecia˛tkowski4,6 •
Wiesław Wiktor-Jedrzejczak1 • Grzegorz W. Basak1
Received: 12 May 2015 / Accepted: 11 December 2015 / Published online: 9 March 2016
Ó The Author(s) 2016. This article is published with open access at Springerlink.com
Abstract Colonization of the gastrointestinal tract with
multidrug-resistant (MDR) bacteria is a consequence of gut
dysbiosis. We describe the successful utilization of fecal
microbiota transplantation to inhibit Klebsiella pneumo-
niae MBL? and Escherichia coli ESBL? gut colonization
in the immunocompromised host as a novel tool in the
battle against MDR microorganisms.
ClinicalTrials.gov identifier NCT02461199.
Keywords Fecal microbiota transplantation
Antibiotic-resistant bacteria Gut colonization
Suppressor Properties of Human CD8+CD282 T Cells in Mixed
Leukocyte Reaction are not Affected by CsA and RAPA
Anna Korecka-Polak1 • Katarzyna Bocian1 • Maria Pacho´wka1 •
Agnieszka Jałbrzykowska2 • Gra_zyna Korczak-Kowalska1,2
Received: 18 August 2015 / Accepted: 11 January 2016 / Published online: 26 February 2016
Ó L. Hirszfeld Institute of Immunology and Experimental Therapy, Wroclaw, Poland 2016
Abstract Human CD8?CD28– T suppressor cells were
previously shown to be involved in the control of the
immune response to transplanted allografts. It seems
essential to examine how immunosuppressive drugs influ-
ence these cells. However, the CD8?CD28– population
contains both suppressor (Ts) and cytotoxic (Tc) T cells,
and the phenotype of the Ts subpopulation has not been
identified explicitly. It is proposed that the transcription
factor FOXP3 may be helpful in distinguishing the Ts and
Tc subpopulations. The aim of this study was to evaluate
the influence of the immunosuppressive drugs cyclosporine
A (CsA) and rapamycin (RAPA) on the level, suppressor
properties, and phenotype of human CD8?CD28– T cells
in vitro. The model used was the mixed leukocyte reaction
performed with peripheral blood mononuclear cells from
healthy volunteers. It was observed that CD8?CD28– T
cells from cultures with CsA or RAPA had similar sup-
pressor properties to cells from control cultures, although
the drugs influenced the expression of FOXP3. CsA and
RAPA did not interfere with the suppressor properties of
human CD8?CD28– T cells in vitro, although they affec-
ted the expression of the FOXP3 molecule.
Keywords Cyclosporine A CD8?CD28– FOXP3
Rapamycin Suppressor T cells
Possible Role of HLA–G, LILRB1 and KIR2DL4 Gene
Polymorphisms in Spontaneous Miscarriage
Izabela Nowak1 • Andrzej Malinowski2 • Ewa Barcz3 • Jacek R. Wilczyn´ ski4 •
Marta Wagner1 • Edyta Majorczyk1,5 • Hanna Motak-Pochrze˛st5,6 •
Małgorzata Banasik7 • Piotr Kus´nierczyk1
Received: 20 October 2015 / Accepted: 28 January 2016 / Published online: 14 March 2016
Ó The Author(s) 2016. This article is published with open access at Springerlink.com
Abstract The KIR2DL4 receptor and its ligand HLA-G
are considered important for fetal-maternal immune toler-
ance and successful pregnancy. The absence of a particular
variant of KIR2DL4 might be a bad prognostic factor for
pregnancy outcome. However, it could be compensated by
the presence of the respective LILRB1 allele. Therefore,
we investigated the KIR2DL4, LILRB1 and HLA–G poly-
morphisms in 277 couples with spontaneous abortion and
219 control couples by HRM, PCR-SSP and RFLP
methods. We found a protective effect of women’s
heterozygosity in –716 HLA–G (p = 0.0206) and LILRB1
(p = 0.0131) against spontaneous abortion. Surprisingly,
we observed more 9A/10A genotypes of KIR2DL4 gene
carriers in the group of male partners from the miscarriage
group in comparison to the men from the control group
(p = 0.0288). Furthermore, there was no association of
women’s KIR2DL4 polymorphism with susceptibility to
spontaneous abortion. Multivariate analysis indicated that
women’s –716 HLA–G and LILRB1 and men’s KIR2DL4
9A/10A are important in terms of the protection or sus-
ceptibility to miscarriage, respectively (p = 0.00968). In
conclusion, a woman’s heterozygosity in HLA–G and
LILRB1 might be an advantage for a success of reproduc-
tion, but the partner’s heterozygosity in 9A/10A KIR2DL4
alleles might not.
Keywords HLA-G LILRB1 Linkage disequilibrium
KIR2DL4 Spontaneous abortion
Intranasal Administration of Type V Collagen Reduces Lung
Carcinogenesis through Increasing Endothelial and Epithelial
Apoptosis in a Urethane-Induced Lung Tumor Model
Edwin Roger Parra1 • Renata Antunes Alveno1 • Carolina Brito Faustino1 •
Paula Yume Sato Serzedello Correˆa1 • Camilla Mutai Vargas1 • Jymenez de Morais2 •
Maristela Peres Rangel1 • Ana Paula Pereira Velosa2 • Alexandre Todorovic Fabro1 •
Walcy Rosolia Teodoro2 • Vera Luiza Capelozzi1
Received: 23 June 2015 / Accepted: 30 November 2015 / Published online: 28 March 2016
Ó L. Hirszfeld Institute of Immunology and Experimental Therapy, Wroclaw, Poland 2016
Abstract Type V collagen (Col V) is a ‘‘minor’’ com-
ponent of normal lung extracellular matrix, which is
subjected to decreased and abnormal synthesis in human
lung infiltrating adenocarcinoma. We previously reported
that a direct link between low amounts of Col V and
decreased cell apoptosis may favor cancer cell growth in
the mouse lung after chemical carcinogenesis. Moreover,
this collagen species was able to trigger DNA fragmenta-
tion and impair survival of neoplastic cells. In this study,
we have extended our investigation with the aim to obtain
further evidence that the death induced by Col V-treatment
is of the caspase-9 apoptotic type. We used (1) optical and
electron microscopy, (2) quantitation of TUNEL-labeled
cells and (3) analysis of the expression levels of Col V and
selected genes coding for apoptosis-linked factors, by
conventional RT-PCR. BALB/c mice were injected
intraperitoneally with 1.5 g/kg body weight of urethane.
After urethane injection, the animals received intranasal
administration of 20 lg/20 ll of Col V every day during
2 months. We report here that Col V treatment was able to
determine significant increase in Col V protein and gene
expression and in the percentage of TUNEL-positive cells,
to up-regulate caspase-9, resulting in low growth of tumor
cells. Our data validate chemical carcinogenesis as a suit-
able ‘‘in vivo’’ model for further and more detailed studies
on the molecular mechanisms of the death response
induced by Col V in lung infiltrating adenocarcinoma
opening new strategies for treatment.
Keywords Chemically-induced lung carcinogenesis
Molecular biology Apoptosis Type V collagen
Immune response Vascular endothelial growth factor
Imatinib Inhibits the Renewal and Tumorigenicity of CT-26 Colon
Cancer Cells after Cytoreductive Treatment with Doxorubicin
Małgorzata Przybyszewska1 • Joanna Miłoszewska1 • Agnieszka Kotlarz1 •
Paweł Swoboda1 • Kazimiera Pys´niak2 • Wojciech Szczepek3 • Łukasz Kaczmarek3 •
Sergiusz Markowicz1
Received: 26 June 2015 / Accepted: 20 January 2016 / Published online: 8 March 2016
Ó L. Hirszfeld Institute of Immunology and Experimental Therapy, Wroclaw, Poland 2016
Abstract Conventional anti-cancer drugs preferentially
eliminate differentiated cancer cells but those cells that are
spared (i.e. cancer stem cells: CSC), initiate recurrence. We
tested whether drugs that target receptor tyrosine kinases
(RTKs) involved in developmental signaling cascades and
activated in CSC, could be used to silence and/or to
eliminate colorectal cancer cells refractory to conventional
treatment with cytoreductive drugs. A sequential treatment
model was thereby developed with doxorubicin (DOX) and
imatinib. CT-26 mouse colon carcinoma cells were pre-
treated with DOX to select DOX-refractory cells with CSC
properties, which were then subsequently treated with RTK
inhibitor imatinib, where their regrowth was found to be
inhibited. Under both normoxic and hypoxic conditions,
imatinib potently inhibited clonogenicity of DOX-refrac-
tory CT-26 cells. Treatment with DOX did not eliminate
tumorigenic CT-26 cells, since CT-26 cells pre-exposed to
DOX in vitro, when inoculated subcutaneously, induced
tumors in 90 % of mice, as opposed to a 100 % rate in the
case of chemonaive CT-26 cells. In mice inoculated with
chemonaive CT-26 cells, tumor formation was not pre-
vented by imatinib. However, imatinib prevented tumor
formation in 50 % of mice inoculated with CT-26 cells pre-
exposed to DOX in vitro, with the remaining 50 % mice
showing delayed tumor formation. These results suggest
that the sequential use of the drug imatinib, as a drug tar-
geting cancer cells expressing stem cell features after
conventional cytoreductive treatment, is a promising future
strategy for preventing tumor recurrence.
Keywords Colorectal cancer Stemness Imatinib
Doxorubicin Drug resistance PDGFRs
Sequential therapy
Pro-Cognitive Properties of the Immunomodulatory Polypeptide
Complex, Yolkin, from Chicken Egg Yolk and Colostrum-Derived
Substances: Analyses Based on Animal Model of Age-Related
Cognitive Deficits
Marta Lemieszewska1 • Marta Jakubik-Witkowska1 • Bartłomiej Stan´ czykiewicz1 •
Aleksandra Zambrowicz2 • Agnieszka Zabłocka3 • Antoni Polanowski2 •
Tadeusz Trziszka2 • Joanna Rymaszewska1
Received: 19 June 2015 / Accepted: 11 December 2015 / Published online: 14 March 2016
Ó The Author(s) 2016. This article is published with open access at Springerlink.com
Abstract The study aimed to assess the effect of the
polypeptide Y complex (Yolkin), isolated from chicken
egg yolk, on behavioural and cognitive functions. It also
aimed to compare this activity with colostrum-derived
substances (Colostrinin, Coloco), which have a confirmed
impact on learning and memory. In the study, the effect of
Yolkin, administered to rats of different ages, who per-
formed various tasks involving spatial and episodic
memory, motor functions and exploratory behavior, was
assessed. The experiment was carried out in rats which
were 6 and 12 months old. Two different doses of the
studied specimens based on previous comparative studies
and two different routes of administration (oral and
retroperitoneal) were used. A series of behavioural tests
were carried out, including an open field test, a novel object
recognition test and a Morris water maze. They were used
to evaluate the impact of the studied specimen on
improving locomotor function and exploratory behaviour,
preventing their decline and assess the functioning of epi-
sodic and spatial memory in aging rats. The administration
of Yolkin gave distinct effects compared to colostrum-
derived substances, although confirmed its suggested pro-
cognitive action. Therefore, it may be used to enhance
cognitive functions and inhibit the progression of dementia
in the course of neurodegenerative disorders.
Keywords Immunomodulatory peptides
Chicken egg yolk Colostrinin Coloco Learning
Memory Dementia Age-related disorders
Cognitive deficits
Does TSH Trigger the Anti-thyroid Autoimmune Processes?
Observation on a Large Cohort of Naive Patients with Thyroid
Hemiagenesis
Ewelina Szczepanek-Parulska1 • Ariadna Zybek-Kocik1 • Kosma Wolin´ ski1 •
Barbara Czarnocka2 • Marek Ruchała1
Received: 15 July 2015 / Accepted: 10 November 2015 / Published online: 14 March 2016
Ó The Author(s) 2016. This article is published with open access at Springerlink.com
Abstract Thyroid hemiagenesis (THA) is a rare abnor-
mality characterized by the absence of one thyroid lobe.
Elevated thyroid stimulating hormone (TSH) level and
higher incidence of thyroid diseases were reported in THA.
The aim of the study is to evaluate the thyroid autoim-
munity incidence in patients with THA and influence of
higher than average TSH level on thyroid volume (TV) and
its change with age. The study included a group of naive
patients with THA and a control group of subjects with
bilobate thyroid. All patients underwent clinical examina-
tion, thyroid ultrasound, scintiscan and laboratory tests. In
the studied and control group the presence of thyroid
autoantibodies (TAb) was evaluated. The THA group
consisted of 65 patients. In THA group 53.85 % of patients
were positive for TAb. Patients with positive TAb were
older (46.0 ± 18.3 years) than those with negative
(35.0 ± 19.8 years); p = 0.02. The incidence of TAb was
lower in controls (13.85 %, p \ 0.0001). In the study
group, positive correlation between the age and TV
(r = 0.46, p = 0.0001), and negative correlations between
the age and TSH level (r = –0.31, p = 0.01), and TSH
concentration and TV (r = –0.35, p = 0.004) were found.
In a subgroup of 30 patients with THA negative for TAb,
even stronger correlations were observed. The median
single lobe volume and median TSH level were higher in
patients with THA when compared to controls (13.60 vs
8.20 ml, p \ 0.0001; 3.23 vs 1.48 lU/ml, p \ 0.0001,
respectively). Patients with THA constitute an in vivo
model of long-term thyroid TSH overstimulation. Further
studies are needed to reveal, whether TSH overstimulation
may be the trigger for thyroid autoimmunity.
Keywords Thyroid Autoimmunity
Hormone secretion Nodular goiter
Adipose-Derived Stem Cells as a Tool in Cell-Based Therapies
Anna Bajek1 • Natalia Gurtowska1 • Joanna Olkowska1 • Lukasz Kazmierski1 •
Malgorzata Maj1 • Tomasz Drewa1,2
Received: 30 September 2015 / Accepted: 20 January 2016 / Published online: 13 May 2016
Ó The Author(s) 2016. This article is published with open access at Springerlink.com
Abstract Recent development in stem cell isolation
methods and expansion under laboratory conditions create
an opportunity to use those aforementioned cells in tissue
engineering and regenerative medicine. Particular attention
is drawn towards mesenchymal stem cells (MSCs) being
multipotent progenitors exhibiting several unique charac-
teristics, including high proliferation potential, self-
renewal abilities and multilineage differentiation into cells
of mesodermal and non-mesodermal origin. High abun-
dance of MSCs found in adipose tissue makes it a very
attractive source of adult stem cells for further use in
regenerative medicine applications. Despite immunomod-
ulating properties of adipose-derived stem cells (ASCs) and
a secretion of a wide variety of paracrine factors that
facilitate tissue regeneration, effectiveness of stem cell
therapy was not supported by the results of clinical trials.
Lack of a single, universal stem cell marker, patient-to-
patient variability, heterogeneity of ASC population com-
bined with multiple widely different protocols of cell
isolation and expansion hinder the ability to precisely
identify and analyze biological properties of stem cells.
The above issues contribute to conflicting data reported in
literature. We will review the comprehensive information
concerning characteristic features of ASCs. We will also
review the regenerative potential and clinical application
based on various clinical trials.
Keywords Adipose-derived stem cells Adipose tissue
Clinical trials Regenerative medicine
Role of CX3CL1 in Diseases
WangMi Liu1 • Libo Jiang1 • Chong Bian1 • Yun Liang1 • Rong Xing1 •
Mumingjiang Yishakea1 • Jian Dong1
Received: 2 September 2015 / Accepted: 21 February 2016 / Published online: 20 April 2016
Ó L. Hirszfeld Institute of Immunology and Experimental Therapy, Wroclaw, Poland 2016
Abstract Chemokines are a family of small 8–10 kDa
inducible cytokines. Initially characterized as chemotactic
factors, they are now considered to affect not just cellular
recruitment. CX3CL1 is a unique chemokine that can exist
in a soluble form, as a chemotactic cytokine, or in a
membrane-attached form that acts as a binding molecule.
Recently, the effects of CX3CL1 on diseases, such as
inflammation and cancer, have been supported and con-
firmed by numerous publications. However, due to its dual
effects, CX3CL1 exerts numerous effects on pathophysio-
logical conditions that have both negative and positive
consequences on pathogenesis and outcome. This review
article summarizes the important scientific and clinical data
that now point to a critical role for CX3CL1 in diseases.
Keywords CX3CL1 Chemokine Disease
Increased Granulocyte Heparanase Activity in Neutrophils
from Patients with Lupus Nephritis and Idiopathic Membranous
Nephropathy
Maciej Szymczak1 • Jakub Kuz´niar1 • Wacław Kopec´1 • Marcelina _Zabin´ ska1 •
Zofia Marchewka2 • Katarzyna Kos´cielska-Kasprzak1 • Marian Klinger1
Received: 13 June 2015 / Accepted: 19 January 2016 / Published online: 18 April 2016
Ó The Author(s) 2016. This article is published with open access at Springerlink.com
Abstract Heparanase is a b-glucuronidase that cleaves
sugar chains of heparan sulfate proteoglycans. It is believed
that heparanase may be involved in the pathogenesis of
proteinuria. The aim of this study was to assess the sig-
nificance of heparanase in the pathogenesis of particular
glomerulonephritis types. The evaluation of heparanase
activity in serum, urine, and granulocytes and superoxide
dismutase (SOD) activity in granulocytes of patients with
lupus nephritis (n = 17), membranous nephropathy
(n = 11), IgA nephropathy (n = 12), focal and segmental
glomerulosclerosis (n = 18), mesangiocapillary glomeru-
lonephritis (n = 12) and in 19 healthy volunteers were
performed. The heparanase activity in granulocytes of
patients with lupus nephritis and membranous nephropathy
was higher than heparanase activity in granulocytes in the
control group (p = 0.02 in both cases). This is the first
observation of this phenomenon. There was no difference
between SOD activity in granulocytes of patients with all
assessed types of glomerulonephritis and the control group.
A positive correlation between heparanase activity in urine
and double-strain DNA antibodies (r = 0.51; p = 0.04),
and reverse correlations between heparanase in urine and
hemolytic activity of the complement (r = –0.57;
p = 0.03) in the lupus nephritis group, and between hep-
aranase activity in granulocytes and serum total protein
level (r = –0.69; p = 0.02) in membranous nephropathy
were observed. Increase in heparanase activity without
changes in superoxide dismutase activity in the granulo-
cytes from patients with lupus nephritis and membranous
nephropathy was observed. It may be used as one of the
markers of these disease activities.
Keywords Heparanase Granulocytes
Glomerular diseases
The Role of TRAF4 and B3GAT1 Gene Expression in the Food
Hypersensitivity and Insect Venom Allergy in Mastocytosis
Aleksandra Go´rska1 • Marta Gruchała-Niedoszytko2 • Marek Niedoszytko1 •
Agnieszka Maciejewska3 • Marta Chełmin´ ska1 • Marcin Skrzypski4 •
Bartosz Wasa˛g5 • Małgorzata Kaczkan2 • Magdalena Lange6 • Bogusław Nedoszytko6 •
Ryszard Pawłowski3 • Sylwia Małgorzewicz2 • Ewa Jassem1
Received: 1 July 2015 / Accepted: 27 January 2016 / Published online: 16 April 2016
Ó The Author(s) 2016. This article is published with open access at Springerlink.com
Abstract Mastocytosis is an uncommon disease classi-
fied as a myeloproliferative neoplasm, however, its
symptoms are broad and place patients at crossroads
between dermatology, hematology and allergology.
Patients with mastocytosis often suffer from symptoms
resulting from the activation and release of mediators from
the mast cells, such as generalized itching, redness, head-
ache, abdominal cramps, diarrhea, bone pain or arthritis,
hypotension and shock. The possible severe, fatal or near
fatal reactions caused by food hypersensitivity are reasons
for the research focused on marker identification. The aim
of the study was to analyse the gene expression differences
in mastocytosis patients with and without food and drug
hypersensitivity and insect venom allergy (IVA). A total of
57 Caucasian patients with mastocytosis were studied
[median age 41.8; range 18–77 years; 15 (26.3 %) males
and 42 (73.7 %) females]. Quantitative RT-PCRs of 11
genes plus ribosomal 18S RNA were run. Symptoms of
food hypersensitivity were found in 12 patients (21 %),
including 3 patients (13 %) with cutaneous mastocytosis
(CM), and 9 (28 %) with indolent systemic mastocytosis
(ISM). IVA was confirmed in 13 patients (22.8 %)
including 6 patients (10.5 %) with CM, and 7 patients
(12.3 %) with ISM. Drug hypersensitivity was diagnosed
in 10 patients (17.5 %). Significant differences in the gene
expression were found for TRAF4 (p = 0.008) in the
comparison of the mastocytosis patients with and without
concomitant food hypersensitivity. Furthermore significant
differences were found in gene expression for B3GAT1
(p = 0.003) in patients with IVA compared to patients
without insect sting anaphylaxis in the medical history. The
expression of studied genes did not differ according to the
presence of drug hypersensitivity. The TRAF4 expression
was higher in mastocytosis patients with food hypersensi-
tivity in their medical history, the B3GAT1 expression was
lower in mastocytosis patients with IVA in history.
Keywords Mastocytosis Food allergy
Food hypersensitivity Drug hypersensitivity
Insect sting anaphylaxis
Erratum to: Pro-Cognitive Properties of the Immunomodulatory
Polypeptide Complex, Yolkin, from Chicken Egg Yolk
and Colostrum-Derived Substances: Analyses Based on Animal
Model of Age-Related Cognitive Deficits
Marta Lemieszewska1 • Marta Jakubik-Witkowska1 • Bartłomiej Stan´ czykiewicz1 •
Aleksandra Zambrowicz2 • Agnieszka Zabłocka3 • Antoni Polanowski2 •
Tadeusz Trziszka2 • Joanna Rymaszewska1
Published online: 9 May 2016
Ó L. Hirszfeld Institute of Immunology and Experimental Therapy, Wroclaw, Poland 2016
Erratum to: Arch. Immunol. Ther. Exp.
DOI 10.1007/s00005-016-0392-z
The author would like to correct the following errors in the
online publication of the article:
In page 1, under the Abstract section, line 14 should read as
‘‘Two different doses of the studied specimens based on
previous comparative studies and two different routes of
administration (oral and intraperitoneal) were used’’.
Under the Introduction heading, line 3 should read as
‘‘Cognitive disorders affecting memory and learning abil-
ities normally occur in the process of aging, mild cognitive
impairment and in Alzheimer’s disease (Lindeboom and
Weinstein 2004; Pepeu 2004).’’
Under the Introduction heading, line 32 should read as
‘‘Positive results of preliminary clinical trials on patients
with Alzheimer’s disease, in which from a total of 15
patients receiving oral CLN at a dose of 100 lg, eight
improved according to the MiniMental State Examination,
while disease symptoms stabilized in the remaining seven
patients (Leszek et al. 1999).’’
The corrected Keywords section should read as follows:
Keywords Immunomodulatory peptides Chicken egg
yolk Colostrinin Coloco Learning Memory Dementia
Age-related disorders Cognitive deficits Yolkin
CD4+CD25+CD1272 and CD4+CD25+Foxp3+ Regulatory T Cell
Subsets in Mediating Autoimmune Reactivity in Systemic Lupus
Erythematosus Patients
Marcelina _Zabin´ ska1 • Magdalena Krajewska1 • Katarzyna Kos´cielska-Kasprzak1 •
Katarzyna Jakuszko1 • Dorota Bartoszek1 • Marta Myszka1 • Marian Klinger1
Received: 4 August 2015 / Accepted: 10 January 2016 / Published online: 7 May 2016
Ó The Author(s) 2016. This article is published with open access at Springerlink.com
Abstract The available clinical as well as experimental
studies implicate participation of T regulatory (Treg) subsets
in the pathogenesis and course of systemic lupus erythe-
matosus (SLE). Introduction of the CD4?CD25?CD127– and
CD4?CD25?Foxp3? regulatory subpopulations analysis into
immunological processes assessment and disease activation
prognosis in patients with lupus nephritis (LN) may improve
monitoring of disease activity and enable an early, and thus
more effective, therapeutic treatment. The main goal of the
study was to investigate whether the quantitative changes of
Treg subpopulations are related to the clinical status of
patients with LN. Fifty-four adult SLE patients divided into
two groups according to their SLEDAI and renal SLEDAI
scores were enrolled into the study. Subpopulations of
CD4?CD25?CD127– and CD4?CD25?Foxp3? phenotypes
were determined by flow cytometry. The control group had
higher absolute number of CD4?CD25?Foxp3? cells com-
pared with the study group (p \ 0.001). Also, significant
inverse correlation in the absolute number of CD4?CD25?–
Foxp3? cells and SLEDAI score was observed. There were
significant differences in the percentage and absolute number
of CD4?CD25?Foxp3? lymphocytes between active and
non-active LN groups. The study group had statistically lower
values of CD4?CD25?CD127– cells, both in the percentage
(p \ 0.001) as well as their absolute number (p = 0.014)
compared to the control group. There were also statistically
significant positive correlations between the absolute number
of CD4?CD25?CD127– and CD4?CD25?Foxp3? Tregs. In
conclusion: (1) reduction in the number of regulatory CD4?–
CD25?Foxp3? cells is a promising indicator of the activity of
SLE, particularly of renal involvement; (2) determination of
the number of regulatory cells using the CD4?CD25?–
CD127– phenotype is unreliable in patients with SLE.
Keywords Systemic lupus erythematosus
Lupus nephritis Regulatory cells Flow cytometry
Humoral and Cellular Autoreactivity to Epidermal Proteins
in Atopic Dermatitis
Cristia´n Navarrete-Dechent1 • Guillermo Pe´rez-Mateluna2 • Sergio Silva-Valenzuela1 •
Cristia´n Vera-Kellet1 • Arturo Borzutzky2,3
Received: 25 August 2015 / Accepted: 8 March 2016 / Published online: 4 May 2016
Ó L. Hirszfeld Institute of Immunology and Experimental Therapy, Wroclaw, Poland 2016
Abstract Atopic dermatitis (AD), a chronic relapsing
inflammatory disease of the skin, is an important public
health concern affecting 10–20 % of children worldwide.
The etiology and pathogenesis of AD involve the interplay
of genetic and environmental factors, including abnor-
malities in skin integrity and a skewed immune system
usually driven by a Th2 phenotype in childhood with a
switch to Th1 in the chronic phase of disease. Children and
adults with AD commonly have elevated IgE levels
directed to multiple different antigens, including aeroal-
lergens, food allergens, and microbial proteins. IgE
targeting self-antigens from epidermal proteins have been
detected in up to 91 % of patients, particularly in severe
persistent AD. It has been suggested that the occurrence of
autoreactivity develops in early childhood. However, it is
not clear yet if autoreactive IgEs in patients with AD are
pathogenic or just an epiphenomenon. The fact that these
autoantibodies are associated with severity and are not
present in other allergic or skin diseases favors the
pathogenicity of IgE-mediated autoreactivity in AD. In this
review, we evaluate the pathogenesis of AD and the
emerging role of autoreactivity to various keratinocyte
antigens involving both the humoral and cellular compo-
nents of the immune system.
Keywords Autoimmunity Epidermis
Immunoglobulin E Malassezia
Allergen-Associated Immunomodulators: Modifying Allergy
Outcome
Cristina Go´mez-Casado1 • Araceli Dı´az-Perales1
Received: 22 October 2015 / Accepted: 21 February 2016 / Published online: 13 May 2016
Ó L. Hirszfeld Institute of Immunology and Experimental Therapy, Wroclaw, Poland 2016
Abstract The prevalence of allergies is increasing since
mid twentieth century; however the underlying causes of
this increase are not fully clear. Understanding the mech-
anism by which a harmless protein becomes an allergen
provides us with the basis to prevent and treat these dis-
eases. Although most studies on allergen immunogenicity
have traditionally focused on structural properties of the
proteins, it is increasingly clear that allergenicity cannot be
determined only based on structural features of the aller-
genic proteins. In fact, allergens do not encounter
human facings as isolated molecules but contained in
complex mixtures of proteins, carbohydrates and lipids,
such as pollen grains or foods. As a result, attention has
lately been directed to examine whether allergen-associ-
ated molecules exhibit immune-regulatory properties. The
present review aims to illustrate some examples of how
non-protein molecules accompanying the allergen can
modulate allergic responses.
Keywords Immunomodulator Allergen LPS
Particles Carbohydrates Lipids
High Efficacy of Methotrexate in Patients with Recurrent
Idiopathic Acute Anterior Uveitis: a Prospective Study
Artur Bachta1 • Bartłomiej Kisiel1 • Mateusz Tłustochowicz2 • Anna Raczkiewicz1 •
Marek Re˛kas2 • Witold Tłustochowicz1
Received: 14 October 2015 / Accepted: 21 February 2016 / Published online: 11 May 2016
Ó L. Hirszfeld Institute of Immunology and Experimental Therapy, Wroclaw, Poland 2016
Abstract To evaluate prospectively the efficacy of
methotrexate (MTX) in the treatment of recurrent idio-
pathic acute anterior uveitis (RIAAU). Nineteen out of 22
RIAAU patients completed the study (two patients with-
drew their consent shortly after study initiation, one patient
discontinued after 4 weeks because of the adverse effects).
All patients were treated with MTX in a starting dose of
15 mg/week, increased to target dose of 25 mg/week after
4 weeks. In patients taking systemic corticosteroids (CS)
the dose was gradually tapered (by 2.5 mg every week)
until discontinuation. The mean follow-up period was
3.3 years (19–59 months). Sixteen patients (84 %)
remained flare-free on MTX therapy. In the remaining
three patients the mean interval between flares increased
from 4.8 to 18.3 months. Systemic CS were tapered off in
all patients. The number of acute anterior uveitis flares in
the whole cohort decreased from 2.12 to 0.11/patient-year
(p \ 0.0001). All flares observed on MTX therapy occur-
red in HLA-B27-positive patients. MTX dosed at 25 mg/
week is highly effective in the treatment of RIAAU.
Keywords Uveitis Inflammation Immunotherapy
Methotrexate
A Special Connection between cd T Cells and Natural Antibodies?
Willi K. Born1,2 • Yafei Huang3 • Wanjiang Zeng4 • Raul M. Torres2 •
Rebecca L. O’Brien1,2
Received: 21 January 2016 / Accepted: 4 April 2016 / Published online: 27 May 2016
Ó L. Hirszfeld Institute of Immunology and Experimental Therapy, Wroclaw, Poland 2016
Abstract Natural antibodies (NAbs) play an important
role in early host defense, autophagy and tissue remodel-
ing, and in immune regulation. They arise spontaneously
(without specific immunization), and are already present at
birth. NAbs are produced by B1 B cells, MZ B cells and
other B cell types. They include all major Ig subclasses but
IgM antibodies are prevalent, especially early in develop-
ment. NAbs may be poly-specific, recognize particular
auto-antigens, or detect neo-determinants such as those
exposed during apoptosis or generated by oxidation. NAbs
do not require cognate T cell help but depend on soluble
mediators produced by T cells. Our recent studies suggest
that cd T cells may have a special relationship with NAbs,
and play a prominent role in their regulation, in part
through the fine-tuning of IL-4 levels. The spontaneously
activated state of these cells likely enables their cytokine
production and other functions in the absence of external
stimulation. Ontogenetically, the earlier arising cd T cells
are better positioned than ab T cells to shape the devel-
oping repertoire of NAbs. Intriguingly, ligand specificities
of NAbs and cd T cell receptors appear to be overlapping,
perhaps allowing cd cognate help for certain NAb speci-
ficities. Via NAbs, cd T cells could exert a regulatory
influence on numerous processes in health and disease.
Keywords Natural antibodies cd T cells
Immune system development
Program Death-1 Suppresses Autoimmune Arthritis by Inhibiting
Th17 Response
Lifen Yang1,2 • Guilin Qiao2 • Yassir Hassan2 • Zhenping Li2 • Xiaoqing Zhang1 •
Huimin Kong1 • Weimin Zeng4 • Fei Yin1 • Jian Zhang2,3
Received: 10 September 2015 / Accepted: 14 March 2016 / Published online: 19 May 2016
Ó L. Hirszfeld Institute of Immunology and Experimental Therapy, Wroclaw, Poland 2016
Abstract Program death-1 (PD-1) is a co-inhibitory
receptor inducibly expressed on activated T cells. PD-1 has
been reported to be associated with the development of
several autoimmune diseases including rheumatoid arthri-
tis, but the precise cellular and molecular mechanisms have
not been fully elucidated. To study the role of PD-1 in the
pathogenesis of rheumatoid arthritis and the possible
underlying mechanisms, we performed collagen-induced
arthritis (CIA) in C57BL/6 mice. Here, we show that PD-1
deficiency leads to the development of severe CIA in mice.
When analyzing T cells from CIA mice ex vivo, we noticed
aberrant antigen-specific Th17 responses in mice lacking
PD-1. This is possibly due to deregulated activation of
PKC-h and Akt. In support of this notion, treating Pd-
cd1–/– mice with an inhibitor of PI3-kinase that is
upstream of PKC-h and Akt significantly suppressed the
disease severity. Therefore, our data indicate that PD-1
dampens antigen-specific Th17 response, thus inhibiting
the disease.
Keywords Program death-1 Collagen-induced arthritis
Th17 PI3-kinase Akt
Transcription Factor NF-jB: An Update on Intervention
Strategies
Arvind Panday1,2 • Maria Eugenia Inda3 • Prathyusha Bagam4 • Malaya K. Sahoo5 •
Diana Osorio1 • Sanjay Batra1,4
Received: 10 December 2015 / Accepted: 14 April 2016 / Published online: 28 May 2016
Ó L. Hirszfeld Institute of Immunology and Experimental Therapy, Wroclaw, Poland 2016
Abstract The nuclear factor (NF)-jB family of tran-
scription factors are ubiquitous and pleiotropic molecules
that regulate the expression of more than 150 genes
involved in a broad range of processes including inflam-
mation, immunity, cell proliferation, differentiation, and
survival. The chronic activation or dysregulation of NF-jB
signaling is the central cause of pathogenesis in many
disease conditions and, therefore, NF-jB is a major focus
of therapeutic intervention. Because of this, understanding
the relationship between NF-jB and the induction of var-
ious downstream signaling molecules is imperative. In this
review, we provide an updated synopsis of the role of NF-
jB in DNA repair and in various ailments including car-
diovascular diseases, HIV infection, asthma, herpes
simplex virus infection, chronic obstructive pulmonary
disease, and cancer. Furthermore, we also discuss the
specific targets for selective inhibitors and future thera-
peutic strategies.
Keywords NF-jB Asthma COPD HIV HSV
Cancer DNA damage
Alpha-Ketoglutarate as a Molecule with Pleiotropic Activity:
Well-Known and Novel Possibilities of Therapeutic Use
Barbara Zdzisin´ ska1 • Aleksandra _Zurek1 • Martyna Kandefer-Szerszen´ 1
Received: 21 December 2015 / Accepted: 22 February 2016 / Published online: 20 June 2016
Ó The Author(s) 2016. This article is published with open access at Springerlink.com
Abstract Alpha-ketoglutarate (AKG), an endogenous
intermediary metabolite in the Krebs cycle, is a molecule
involved in multiple metabolic and cellular pathways. It
functions as an energy donor, a precursor in the amino acid
biosynthesis, a signalling molecule, as well as a regulator
of epigenetic processes and cellular signalling via protein
binding. AKG is an obligatory co-substrate for 2-oxoglu-
tarate-dependent dioxygenases, which catalyse
hydroxylation reactions on various types of substrates. It
regulates the activity of prolyl-4 hydroxylase, which con-
trols the biosynthesis of collagen, a component of bone
tissue. AKG also affects the functioning of prolyl
hydroxylases, which, in turn, influences the function of the
hypoxia-inducible factor, an important transcription factor
in cancer development and progression. Additionally, it
affects the functioning of enzymes that influence epigenetic
modifications of chromatin: ten–eleven translocation
hydroxylases involved in DNA demethylation and the
Jumonji C domain containing lysine demethylases, which
are the major histone demethylases. Thus, it regulates gene
expression. The metabolic and extrametabolic function of
AKG in cells and the organism open many different fields
for therapeutic interventions for treatment of diseases. This
review presents the results of studies conducted with the
use of AKG in states of protein deficiency and oxidative
stress conditions. It also discusses current knowledge about
AKG as an immunomodulatory agent and a bone anabolic
factor. Additionally, the regulatory role of AKG and its
structural analogues in carcinogenesis as well as the results
of studies of AKG as an anticancer agent are discussed.
Keywords Alpha-ketoglutarate Antioxidative factor
Dietary supplement Immunomodulatory agent
Bone anabolic agent Anticancer agent
Biomarkers Guided Treatment Strategies in Adult Patients
with Asthma: Ready for the Clinical Field?
Zoi Tsilogianni1 • Polyxeni Ntontsi2 • Andriana I. Papaioannou2 •
Petros Bakakos1 • Stelios Loukides2
Received: 5 January 2016 / Accepted: 14 April 2016 / Published online: 8 June 2016
Ó L. Hirszfeld Institute of Immunology and Experimental Therapy, Wroclaw, Poland 2016
Abstract Asthma is a chronic inflammatory airways dis-
order mainly characterized by heterogeneity. In the more
severe forms, a discordance often exists between symptoms
and inflammation. Difficulty in managing asthma derives
partly from the multiple phenotypes existing and our
inability to recognize them. The use of non-invasive, with
main representative the fraction of exhaled nitric oxide, or
semi-invasive techniques such as induced sputum are
effective tools that can help us to guide asthma treatment.
In the latest years, several serum biomarkers related to
asthmatic inflammation have been used for the better
recognition of asthma sub-phenotypes to achieve opti-
mization of therapy and disease outcome. In patients with
mild–moderate asthma, as well as patients with more
severe asthma, the use of blood eosinophils revealed an
acceptable accuracy for the prediction of airway eosino-
philia indicating that in future studies may facilitate both
individualized treatment and management of asthma. None
of the above techniques have been incorporated in clinical
practice although sputum eosinophils can be used in
patients with severe asthma particularly in specialized
centers with great experience. Of great interest are blood
eosinophils since current data support their role either as
tool for treatment selections or/and as a biomarker of air-
way eosinophilia.
Keywords Asthma Biomarkers Treatment Strategy
Eosinophils Sputum
The Hidden Side of Disodium Cromolyn: from Mast Cell
Stabilizer to an Angiogenic Factor and Antitumor Agent
Anca Maria Cimpean1 • Marius Raica1
Received: 29 November 2015 / Accepted: 9 March 2016 / Published online: 11 June 2016
Ó L. Hirszfeld Institute of Immunology and Experimental Therapy, Wroclaw, Poland 2016
Abstract Scattered data suggested that disodium cro-
molyn, well known as a mast cell stabilizer shows some
effects on tumor cells and tumor-associated newly formed
vascular networks. Most of these studies used tumor cell
lines assessed by in vitro studies. Nor disodium cromolyn
effects on melanoma cell lines were studied yet, neither its
influence on recruited tumor blood vessels or angiogenic
growth factors expression. We designed here a study
regarding disodium cromolyn effects on A375 melanoma
tumor cells implanted on chick embryo chorioallantoic
membrane (CAM) and on blood vessels recruited by the
experimental melanoma in the absence of mast cells,
knowing that within CAM, the existence of mast cells are
not certified yet. We also assessed the role of disodium
cromolyn on the expression of several angiogenic growth
factors. Disodium cromoglycate differentially acts on
tumor cells and blood vessels. Extensive necrotic areas of
experimental melanoma together with an increased number
of peritumor blood vessels were observed in treated spec-
imens as compared with untreated tumors. Disodium
cromolyn inhibited VEGF and PDGF-BB expression, and
had no effects on EG VEGF expression between treated
and non treated specimens in a mast cells free microenvi-
ronment. Our results sustain the direct antitumor effects of
sodium cromolyn and suggest the involvement of several
growth factors in the recruitment of tumor vessels by A375
melanoma tumor cells. The expression of growth factors is
differentially influenced by sodium cromolyn treatment.
Keywords Sodium cromolyn Blood vessels
Tumor cells Chorioallantoic membrane
Epithelial–Mesenchymal Transition in Chronic Rhinosinusitis:
Differences Revealed Between Epithelial Cells from Nasal Polyps
and Inferior Turbinates
Michael Ko¨nnecke1 • Maike Burmeister1 • Ralph Pries1 • Robert Bo¨scke1 •
Karl-Ludwig Bruchhage1 • Hendrik Ungefroren2 • Ludger Klimek3 •
Barbara Wollenberg1
Received: 10 August 2015 / Accepted: 29 April 2016 / Published online: 8 July 2016
Ó L. Hirszfeld Institute of Immunology and Experimental Therapy, Wroclaw, Poland 2016
Abstract The pathogenesis of chronic rhinosinusitis
(CRS) remains unclear to date. The tissue remodeling in
nasal polyps may be the result of inflammatory mediators
and may involve epithelial–mesenchymal transition (EMT)
and EMT-associated features such as cell motility in nasal
epithelial cells (NECs). We determined whether NEC in
nasal polyps of CRS already display features of EMT
in vivo or respond with EMT to growth factor stimulation
in vitro. Nasal polyp tissues expressed both epithelial and
mesenchymal markers. Primary NEC from inferior turbi-
nates and nasal polyps responded to the EMT-inducing
agents transforming growth factor (TGF)-b1 and epidermal
growth factor (EGF) with different expression patterns of
EMT markers (E-cadherin, N-cadherin, Snail, Slug, Twist),
however, only NEC from nasal polyps were susceptible to
TGF-b1 and EGF-dependent cell migration. Our data
suggest that a partial EMT is associated with the patho-
genesis of nasal polyps in CRS patients. Furthermore, we
show for the first time that epithelial cells from both nasal
polyps and inferior turbinates were able to undergo an
EMT-like process following exposure to TGF-b1 or EGF
in vitro but that only NEC from nasal polyps responded
with enhanced cell motility. Our data suggest that NEC
from CRS patients have undergo partial EMT and that this
process may be involved in the pathogenesis of CRS.
Keywords Chronic rhinosinusitis with nasal polyps
CRSwNP Nasal polyps Nasal epithelial cells
Epithelial–mesenchymal transition EMT
Anaphylaxis to IVIG
Sharon Julie Williams1 • Sudhir Gupta1
Received: 8 December 2015 / Accepted: 4 April 2016 / Published online: 13 July 2016
Ó L. Hirszfeld Institute of Immunology and Experimental Therapy, Wroclaw, Poland 2016
Abstract Anaphylactic reactions are a known complica-
tion in some IgA-deficient patients receiving blood or
plasma transfusions. It is of particular interest that ana-
phylaxis has been observed in patients with common
variable immunodeficiency (CVID) who are receiving
intravenous gammaglobulin (IVIG), and in that, although
these patients have an impaired response to common vac-
cines, they retain the ability to produce autoantibodies. In
this study, we review IgA antibodies (both IgG- and IgE-
mediated reactions) in patients with CVID and hypogam-
maglobulinemia, anaphylaxis in antibody immunodeficient
patients receiving IVIG, and proposed mechanisms of
desensitization and prevention of anaphylactic reactions in
immunodeficient patients receiving IVIG. We summarize
and assess the 23 case reports documented in the literature
that have described anaphylactic reactions in immunode-
ficient patients receiving IVIG since 1962 until currently,
and make a comparison of their immunoglobulin levels,
IgG anti-IgA, IgE anti-IgA, concentration of IVIG, IgA
content in IVIG, method used to detect antibodies, length
of treatment, and any subsequent tolerated treatment
documented.
Keywords Anaphylaxis Anaphylactic IVIG
CVID Autoantibodies IgA IgE IgG
Antagonizing Retinoic Acid Receptors Increases Myeloid Cell
Production by Cultured Human Hematopoietic Stem Cells
Geoffrey Brown1 • Aleksandra Marchwicka3 • Alan Cunningham2 •
Kai-Michael Toellner2 • Ewa Marcinkowska3
Received: 26 January 2016 / Accepted: 20 April 2016 / Published online: 13 July 2016
Ó The Author(s) 2016. This article is published with open access at Springerlink.com
Abstract Activities of the retinoic acid receptor (RAR)a
and RARc are important to hematopoiesis. Here, we have
investigated the effects of receptor selective agonists and
antagonists on the primitive human hematopoietic cell lines
KG1 and NB-4 and purified normal human hematopoietic
stem cells (HSCs). Agonizing RARa (by AGN195183) was
effective in driving neutrophil differentiation of NB-4 cells
and this agonist synergized with a low amount (10 nM) of
1a,25-dihydroxyvitamin D3 to drive monocyte differenti-
ation of NB-4 and KG1 cells. Treatment of cultures of
human HSCs (supplemented with stem cell factor ± in-
terleukin 3) with an antagonist of all RARs (AGN194310)
or of RARa (AGN196996) prolonged the lifespan of cul-
tures, up to 55 days, and increased the production of
neutrophils and monocytes. Slowing down of cell differ-
entiation was not observed, and instead, hematopoietic
stem and progenitor cells had expanded in number.
Antagonism of RARc (by AGN205728) did not affect
cultures of HSCs. Studies of CV-1 and LNCaP cells
transfected with RAR expression vectors and a reporter
vector revealed that RARc and RARb are activated by sub-
nM all-trans retinoic acid (EC50–0.3 nM): *50-fold more
is required for activation of RARa (EC50–16 nM). These
findings further support the notion that the balance of
expression and activity of RARa and RARc are important
to hematopoietic stem and progenitor cell expansion and
differentiation.
Keywords Retinoic acid receptor Hematopoiesis
Neutrophils Monocytes All-trans retinoic acid
Agonist Antagonist
Polymorphic Variants 279R and 668Q Augment Activity
of Matrix Metalloproteinase-9 in Breath Condensates of Children
with Asthma
Katarzyna Grzela1 • Wioletta Zago´rska1 • Alicja Krejner2 • Malgorzata Litwiniuk2,3,4 •
Anna Zawadzka-Krajewska1 • Marek Kulus1 • Tomasz Grzela2
Received: 2 October 2015 / Accepted: 9 May 2016 / Published online: 9 July 2016
Ó L. Hirszfeld Institute of Immunology and Experimental Therapy, Wroclaw, Poland 2016
Abstract Matrix metalloproteinase (MMP)-9 is involved
in pathophysiology of asthma, mainly asthma-associated
airway remodeling. Exhaled breath condensates (EBC) of
asthmatics contain increased amounts of MMP-9 with
activity higher, than in healthy controls. The increased
activity of MMP-9 may originate from its excessive pro-
duction and activation, but may also result from variations
in MMP-9 structure, which are determined by single
nucleotide polymorphisms (SNPs). In this pilot study we
aimed to assess the possible influence of two functional
MMP-9 polymorphisms, Q279R and R668Q, on enzymatic
activity of MMP-9, measured in EBC of asthmatic chil-
dren. The concentration and activity of MMP-9 were
analyzed in EBC of 20 children with allergic asthma using
specific standard ELISA and novel immunoenzymatic
activity assay. The SNPs of MMP-9 were assessed using
real-time PCR-based genotyping test. We have found that
MMP-9 concentration in breath condensates of children
with stable asthma was slightly higher in ELISA, than in
the activity assay. Moreover, these results and activity-to-
amount ratio have revealed some relationship with a
presence of specific 279R and/or 668Q MMP-9 gene
variants. Our observation suggests that at least in some
patients MMP-9 hyperactivity may result from genetic
predisposition, determined by polymorphic variants of
MMP-9 gene. Moreover, it supports previous reports pos-
tulating significance of MMP-9 in pathogenesis of asthma.
However, this issue still requires further studies.
Keywords ASTHMA Enzyme activity
Exhaled breath condensate Matrix metalloproteinase
Airway remodeling Single nucleotide polymorphism
In Vivo Cardioprotective Effects and Pharmacokinetic Profile
of N-Propyl Caffeamide Against Ischemia Reperfusion Injury
Yuan-Yuan Cheng1 • Dan Luo1 • Zhengyuan Xia2 • Hung-Fat Tse3 •
Xuechen Li4 • Jianhui Rong1
Received: 28 February 2016 / Accepted: 9 May 2016 / Published online: 1 August 2016
Ó L. Hirszfeld Institute of Immunology and Experimental Therapy, Wroclaw, Poland 2016
Abstract Caffeic acid derivatives constitute a class of
potent anti-inflammatory and cardioprotective drug can-
didates. We recently synthesized a new caffeic acid
derivative N-propyl caffeamide (PCA). Our pilot experi-
ments demonstrated that PCA enhanced the survival of rat
cardiomyocyte H9c2 cells against oxygen glucose depri-
vation and reoxygenation challenge in a concentration-
dependent manner. Interestingly, PCA exhibited better
cardioprotective potential than caffeic acid phenethyl
ester and propyl caffeate. Thus, we hypothesized that
PCA could protect heart against ischemia reperfusion (I/
R) injury in mice. We first determined the stability and
pharmacokinetic profile of PCA in male Sprague–Dawley
rats by ultra-performance liquid chromatography coupled
with UV and MS/MS detections. The stability of PCA in
rat plasma was defined by the half-life of 31.39, 7.19 and
1.37 h in rat plasma at 25, 37 and 60 °C, respectively. To
study the pharmacokinetic profiles, PCA was injected into
male SD rats at the dose of 15 mg/kg via intravenous
bolus administration. PCA showed the elimination half-
life of approximate 235 min in rats. We subsequently
evaluated the cardioprotective potential of PCA in mice
model of myocardial infarction. Our results demonstrated
that PCA effectively reduced infarct size and release of
myocardial enzymes (e.g., CK, CK-MB and LDH). Bio-
chemical analyses suggested that PCA increased the
activities of antioxidant enzymes (e.g., CAT and SOD)
while attenuated lipid peroxidation. Moreover, PCA pro-
foundly reduced the number of apoptotic cells in infarcted
myocardium. Consistently, PCA increased the expression
level of anti-apoptotic protein Bcl2 whereas suppressed
the expression of pro-apoptotic protein Bax in cardiac
tissues. Collectively, PCA appears to be a novel
bioavailable and stable pharmacological treatment for
myocardial infarction.
Keywords N-Propyl caffeamide Bioavailability
Pharmacokinetics Cardioprotection
Ischemia reperfusion injury
Indications to Epigenetic Dysfunction in the Pathogenesis
of Common Variable Immunodeficiency
William Rae1
Received: 12 January 2016 / Accepted: 10 June 2016 / Published online: 2 August 2016
Ó L. Hirszfeld Institute of Immunology and Experimental Therapy, Wroclaw, Poland 2016
Abstract Primary immunodeficiencies (PIDs) are a group
of rare genetic diseases resulting in the impairment of one
or more functions of the human immune system. Common
variable immunodeficiency (CVID) is one of the most
prevalent PIDs, yet despite extensive genetic analysis, most
patients do not have a monogenetic diagnosis. This has led
to the theory that CVID must be a polygenetic condition.
An alternative theory to a monogenetic or polygenetic
underlying cause of CVID is that it is epigenetic phe-
nomena that are causal in the majority of CVID patients. I
will briefly discuss epigenetic regulation in B-cell biology
and development, current examples of epigenetic diseases
causing CVID-like primary antibody deficiencies, and how
these observations may guide future investigation into the
role of epigenetics in CVID.
Keywords Epigenetics B-cell development
Common variable immunodeficiency
Immunotherapy of Sepsis: Blind Alley or Call for Personalized
Assessment?
Miroslav Prucha1 • Roman Zazula2 • Stefan Russwurm3
Received: 18 December 2015 / Accepted: 14 April 2016 / Published online: 24 August 2016
Ó L. Hirszfeld Institute of Immunology and Experimental Therapy, Wroclaw, Poland 2016
Abstract Sepsis is the most frequent cause of death in
noncoronary intensive care units. In the past 10 years,
progress has been made in the early identification of septic
patients and their treatment. These improvements in sup-
port and therapy mean that mortality is gradually
decreasing, however, the rate of death from sepsis remains
unacceptably high. Immunotherapy is not currently part of
the routine treatment of sepsis. Despite experimental suc-
cesses, the administration of agents to block the effect of
sepsis mediators failed to show evidence for improved
outcome in a multitude of clinical trials. The following
survey summarizes the current knowledge and results of
clinical trials on the immunotherapy of sepsis and describes
the limitations of our knowledge of the pathogenesis of
sepsis. Administration of immunomodulatory drugs should
be linked to the current immune status assessed by both
clinical and molecular patterns. Thus, a careful daily
review of the patient’s immune status needs to be intro-
duced into routine clinical practice giving the opportunity
for effective and tailored use of immunomodulatory
therapy.
Keywords Sepsis Immune mechanisms
Immunosuppression Immunomodulatory therapy
Genomics Proteomics
Expression Profiles of Toll-Like Receptors in the Differentiation
of an Infection with Borrelia burgdorferi Sensu Lato Spirochetes
Slawomir Dudek1 • Ewa Zio´łko2 • Magdalena Kimsa-Dudek3 • Krzysztof Solarz4 •
Urszula Mazurek5 • Aleksander Wierzgon´ 6 • Teresa Kokot2 • Małgorzata Muc-Wierzgon´ 2
Received: 11 November 2015 / Accepted: 1 June 2016 / Published online: 7 September 2016
Ó L. Hirszfeld Institute of Immunology and Experimental Therapy, Wroclaw, Poland 2016
Abstract The similarity of Lyme borreliosis to other dis-
eases and its complex pathogenesis present diagnostic and
therapeutic difficulties. The changes that occur at the cel-
lular and molecular levels after a Borrelia sp. infection still
remain poorly understood. Therefore, the present study
focused on the expression of TLR and TLR-signaling genes
in human dermal fibroblasts in the differentiation of an
infection with Borrelia burgdorferi sensu lato spirochetes.
Normal human dermal fibroblasts were cultured with the
spirochetes of Borrelia burgdorferi sensu stricto, Borrelia
afzelii and Borrelia garinii. Total RNA was extracted from
the cells using TRIzol reagent. The analysis of the
expression profiles of TLRs and TLR-related genes was
performed using commercially available oligonucleotide
microarrays of HG-U133A. The GeneSpring 12.0 platform
and significance analysis of microarrays were used for the
statistical analysis of microarray data. The analyses using
the oligonucleotide microarray and QRT-PCR techniques
permitted to identify the genes encoding TLR4 and TLR6 as
specific for infection with B. afzelii and B. burgdorferi
sensu stricto. In turn, TLR3 was only characteristic for an
infection with B. burgdorferi sensu stricto. There were no
changes in the TLR gene expression after infection with B.
garinii. Our findings confirm that Borrelia has a major
effect on fibroblast gene expression. Further characteriza-
tion of changes in gene expression may lead to valuable
insights into the role of the toll-like receptor in the
pathogenesis of Lyme disease and may provide guidelines
for the development of diagnostic markers for an infection
with a particular Borrelia genospecies. Moreover, this will
help to identify better treatment strategies for Lyme
disease.
Keywords Toll-like receptors
Borrelia burgdorferi sensu lato spirochetes
Fibroblasts cultures Oligonucleotide microarray
QRT-PCR
Autoreactive IgE in Chronic Spontaneous/Idiopathic Urticaria
and Basophil/Mastocyte Priming Phenomenon, as a Feature
of Autoimmune Nature of the Syndrome
Bernard Panaszek1 • Robert Pawłowicz1 • Je˛drzej Grzegrzo´łka2 • Andrzej Obojski1
Received: 1 September 2015 / Accepted: 20 June 2016 / Published online: 31 August 2016
Ó L. Hirszfeld Institute of Immunology and Experimental Therapy, Wroclaw, Poland 2016
Abstract Recent years of research have shed a new light on
the role of IgE in immune reactions. It seems to be more than
just a contribution to immediate type of allergic response. It
appears that monomeric IgE may enhance mast cell activity
without cross-linking of FceRI by IgE specific allergen or
autoreactive IgG anti-IgE antibodies. Monomeric IgE
molecules are heterogeneous concerning their ability to
induce survival and activation of mast cells only by binding
the IgE to FceRI, but not affecting degranulation of cells. It
also turned out that IgE may react to autoantigens occurring
in the blood not only in chronic spontaneous urticaria (CSU)
but also in other autoimmune diseases. The aforementioned
phenomena may promote the activity of mast cells/basophils
in CSU that easily degranulate when influenced by various
inner (autoreactive IgG against IgE and FceRI, autoreactive
IgE for self-antigens) and outer factors (cold, heat, pressure)
or allergens. These findings forced the new approach to the
role of autoimmunity, self-antigens and IgE autoantibodies
in the pathology of CSU. CSU put in the scheme of autore-
active IgG and autoreactive IgE seems to be either a kind of
an autoimmune disease or a clinical manifestation of some
other defined autoimmune diseases or both.
Keywords Autoreactive IgE Chronic urticaria
Self-antigens Mastocyte priming Autoimmune reactivity
Hormonal Modulation of Dendritic Cells Differentiation,
Maturation and Function: Implications for the Initiation
and Progress of Systemic Autoimmunity
Juan Pablo Mackern-Oberti1,2,3 • Evelyn L. Jara3 • Claudia A. Riedel4 •
Alexis M. Kalergis3,5,6
Received: 24 March 2016 / Accepted: 4 July 2016 / Published online: 1 September 2016
Ó L. Hirszfeld Institute of Immunology and Experimental Therapy, Wroclaw, Poland 2016
Abstract Hormonal homeostasis is crucial for keeping a
competent and healthy immune function. Several hormones
can modulate the function of various immune cells such as
dendritic cells (DCs) by influencing the initiation of the
immune response and the maintenance of peripheral tol-
erance to self-antigens. Hormones, such as estrogens,
prolactin, progesterone and glucocorticoids may pro-
foundly affect DCs differentiation, maturation and function
leading to either a pro-inflammatory or an anti-inflamma-
tory (or tolerogenic) phenotype. If not properly regulated,
these processes can contribute to the pathogenesis of
autoimmune disease. An unbalanced hormonal status may
affect the production of pro-inflammatory cytokines, the
expression of activating/inhibitory receptors and co-
stimulatory molecules on conventional and plasmacytoid
DCs (pDCs), conferring susceptibility to develop autoim-
munity. Estrogen receptor (ER)-a signaling in conventional
DCs can promote IFN-a and IL-6 production and induce
the expression of CD40, CD86 and MHCII molecules.
Furthermore, estrogen modulates the pDCs response to
Toll-like receptor ligands enhancing T cell priming. During
lupus pathogenesis, ER-a deficiency decreased the
expression of MHC II on pDCs from the spleen. In con-
trast, estradiol administration to lupus-prone female mice
increased the expression of co-stimulatory molecules,
enhanced the immunogenicity and produced large amounts
of IL-6, IL-12 and TNF-a by bone marrow-derived DCs.
These data suggest that estradiol/ER signaling may play an
active role during lupus pathology. Similarly, understand-
ing hormonal modulation of DCs may favor the design of
new therapeutic strategies based on autologous tolerogenic
DCs transfer, especially in sex-biased systemic autoim-
mune diseases. In this review, we discuss recent data
relative to the role of different hormones (estrogen, pro-
lactin, progesterone and glucocorticoids) in DC function
during systemic autoimmune pathogenesis.
Keywords Estrogen Prolactin Progesterone
Glucocorticoids Autoimmunity Dendritic cells
Systemic lupus erythematosus
Immunosensors for Biomarker Detection in Autoimmune Diseases
Xuezhu Zhang1 • Amarayca Zambrano1 • Zuan-Tao Lin1 • Yikun Xing1 •
Justin Rippy1 • Tianfu Wu1
Received: 30 March 2016 / Accepted: 4 July 2016 / Published online: 3 September 2016
Ó L. Hirszfeld Institute of Immunology and Experimental Therapy, Wroclaw, Poland 2016
Abstract Autoimmune diseases occur when the immune
system generates proinflammatory molecules and autoan-
tibodies that mistakenly attack their own body. Traditional
diagnosis of autoimmune disease is primarily based on
physician assessment combined with core laboratory tests.
However, these tests are not sensitive enough to detect
early molecular events, and quite often, it is too late to
control these autoimmune diseases and reverse tissue
damage when conventional tests show positivity for dis-
ease. It is fortunate that during the past decade, research in
nanotechnology has provided enormous opportunities for
the development of ultrasensitive biosensors in detecting
early biomarkers with high sensitivity. Biosensors consist
of a biorecognition element and a transducer which are able
to facilitate an accurate detection of proinflammatory
molecules, autoantibodies and other disease-causing
molecules. Apparently, novel biosensors could be superior
to traditional metrics in assessing the drug efficacy in
clinical trials, especially when specific biomarkers are
indicative of the pathogenesis of disease. Furthermore, the
portability of a biosensor enables the development of point-
of-care devices. In this review, various types of biomole-
cule sensing systems, including electrochemical, optical
and mechanical sensors, and their applications and future
potentials in autoimmune disease treatment were discussed.
Keywords Biosensor Biomarker
Autoimmune diseases Nanotechnology
The Use of Omalizumab in Food Oral Immunotherapy
Roxane Labrosse1 • Franc¸ois Graham2 • Anne Des Roches1 • Philippe Be´gin1,2
Received: 2 February 2016 / Accepted: 4 July 2016 / Published online: 14 September 2016
Ó L. Hirszfeld Institute of Immunology and Experimental Therapy, Wroclaw, Poland 2016
Abstract Food allergy is an important health issue that
affects up to 8 % of the population. The management of
allergic patients involves allergen avoidance and prompts
the treatment of accidental reactions, as no curative treat-
ment is available so far in routine practice. Oral
immunotherapy (OIT) is a promising therapeutic alterna-
tive, but it is associated with frequent allergic reactions and
cost-effectiveness issues. In hopes of reducing such reac-
tions, a number of trials have used omalizumab, an anti-IgE
monoclonal humanized antibody, as adjunctive therapy in
OIT. The allergens studied in these omalizumab-enabled
OIT trials include peanuts, milk, eggs, or mixes of multiple
foods. In this article, we review the major findings from
these studies and discuss potential benefits and issues
related to omalizumab-enabled OIT. Results from the
previous trials suggest that the use of omalizumab could
potentially lead to safer and more efficient OIT protocols,
by reducing the number and severity of reactions, and
increasing allergen tolerance threshold. While more evi-
dence is needed with regard to the maintenance of the long-
term tolerance after OIT, omalizumab’s potential
immunomodulatory role could be of benefit. More studies
are needed to further document this new indication for
omalizumab.
Keywords Immunotherapy Desensitization
Oral immunotherapy Food allergy Omalizumab
Anti-IgE therapy
Activated and Memory T Lymphocytes in Children with Gaucher
Disease
Asmaa M. Zahran1 • Azza A. Eltayeb2 • Khalid I. Elsayh2 • Khaled Saad2 •
Faisal-Alkhateeb Ahmad2 • Ahmad I. M. Ibrahim2
Received: 17 February 2016 / Accepted: 1 June 2016 / Published online: 16 September 2016
Ó L. Hirszfeld Institute of Immunology and Experimental Therapy, Wroclaw, Poland 2016
Abstract Gaucher disease (GD) is the most prevalent
lysosomal storage disorder. Gaucher disease is associated
with remarkable alterations in the immune system, and GD
patients are more susceptible to infections and are at a
higher risk of developing autoimmune disorders and
malignancies. In a case–control study, we used three-color
flow cytometric immunophenotyping for determination of
the frequency of lymphocyte subpopulations and activated
T lymphocytes among 18 children with GD1 under enzyme
replacement therapy managed in Assiut University Hospi-
tals. We found significant increases in the frequencies of
total lymphocytes, CD19?, CD3?, CD4?, and CD8? in
children with GD1 when compared to healthy control. The
frequencies of activated T lymphocytes (CD3?HLA-DR?),
activated T-helper cells (CD4?HLA-DR?), and activated
T-suppressor/cytotoxic cells (CD8?HLA-DR?) were sig-
nificantly higher in GD1 as compared to healthy children.
Our data show that the increased proportion of activated T
lymphocytes in children with GD1 raises the issue of their
possible involvement in the pathogenesis of the immune
dysfunction seen in these patients. Our data suggested that
the activated T lymphocytes could play a role in the clin-
ical course of GD1. The relationship of these cells to
immune disorders in GD1 children remains to be
determined.
Keywords Activated T lymphocytes Children
Gaucher disease
Clinical Use and Therapeutic Potential of IVIG/SCIG,
Plasma-Derived IgA or IgM, and Other Alternative
Immunoglobulin Preparations
Peter J. Spa¨th1 • Christoph Schneider1 • Stephan von Gunten1
Received: 19 February 2016 / Accepted: 31 August 2016 / Published online: 16 September 2016
Ó L. Hirszfeld Institute of Immunology and Experimental Therapy, Wroclaw, Poland 2016
Abstract Intravenous and subcutaneous immunoglobulin
preparations, consisting of IgG class antibodies, are
increasingly used to treat a broad range of pathological
conditions, including humoral immune deficiencies, as well
as acute and chronic inflammatory or autoimmune disor-
ders. A plethora of Fab- or Fc-mediated immune regulatory
mechanisms has been described that might act separately or
in concert, depending on pathogenesis or stage of clinical
condition. Attempts have been undertaken to improve the
efficacy of polyclonal IgG preparations, including the
identification of relevant subfractions, mild chemical
modification of molecules, or modification of carbohydrate
side chains. Furthermore, plasma-derived IgA or IgM
preparations may exhibit characteristics that might be
exploited therapeutically. The need for improved treatment
strategies without increase in plasma demand is a goal and
might be achieved by more optimal use of plasma-derived
proteins, including the IgA and the IgM fractions. This
article provides an overview on the current knowledge and
future strategies to improve the efficacy of regular IgG
preparations and discusses the potential of human plasma-
derived IgA, IgM, and preparations composed of mixtures
of IgG, IgA, and IgM.
Keywords Immunoglobulin preparations SCIG
IVIG Polyclonal IgG Polyclonal IgA Polyclonal IgM
Host defence Immunomodulation
A Quinone-Containing Compound Enhances Camptothecin-
Induced Apoptosis of Lung Cancer Through Modulating
Endogenous ROS and ERK Signaling
Han-Lin Chou1,2 • Yao Fong3 • Chi-Ku Wei1 • Eing-Mei Tsai4 • Jeff Yi-Fu Chen1 •
Wen-Tsan Chang5,6 • Chang-Yi Wu1,7 • Hurng-Wern Huang2 • Chien-Chih Chiu1,4,7,8
Received: 18 January 2016 / Accepted: 21 July 2016 / Published online: 27 September 2016
Ó L. Hirszfeld Institute of Immunology and Experimental Therapy, Wroclaw, Poland 2016
Abstract The natural compound camptothecin (CPT)
derivatives have widely been used for anti-cancer treat-
ments, including lung cancer. However, many
chemoresistant cancer cells often develop a relatively
higher threshold for inducing apoptosis, causing a limited
efficacy of anti-cancer drugs. Likewise, lung cancer cells
acquire chemoresistance against CPT analogs, such as
irinotecan and topotecan, finally resulting in an unsatisfied
outcome and poor prognosis of lung cancer patients. TFPP
is a quinone-containing compound as a candidate for CPT-
based combination chemotherapy. In this study, we
examined the effect of TFPP and CPT cotreatment on non-
small cell lung cancer (NSCLC) cells. Cell proliferation
and flow cytometry-based Annexin-V/PI staining assays
demonstrated the synergistic effect of TFPP on CPT-in-
duced apoptosis in both NSCLC A549 and H1299 cells.
The results of CPT and TFPP cotreatment cause the reg-
ulation of the ERK-Bim axis and the activation of
mitochondrial-mediated caspase cascade, including cas-
pase-9 and caspase-3. Besides, TFPP significantly
enhanced CPT-induced endogenous reactive oxygen spe-
cies (ROS) in the two NSCLC cells. In contrast, the
treatment of N-acetyl-L-cysteine (NAC), an ROS scav-
enger, rescues the apoptosis of NSCLC cells induced by
TFPP and CPT cotreatment, suggesting that the synergistic
effect of TFPP on CPT-induced anti-NSCLC cells is
through upregulating ROS production. Consequently, our
results suggest that TFPP sensitizes NSCLC towards CPT-
based chemotherapy may act through decreasing the
apoptosis-initiating threshold. Therefore, TFPP may be a
promising chemosensitizer for lung cancer treatment, and
the underlying mechanism warrants further.
Keywords Non-small cell lung cancer Chemoresistance
Apoptosis threshold Camptothecin TFPP
ERK signaling pathway
LL-37 but Not 25-Hydroxy-Vitamin D Serum Level Correlates
with Healing of Venous Leg Ulcers
Alicja Krejner1 • Małgorzata Litwiniuk1,2,3 • Tomasz Grzela1,4
Received: 1 January 2016 / Accepted: 10 June 2016 / Published online: 23 September 2016
Ó The Author(s) 2016. This article is published with open access at Springerlink.com
Abstract Human cathelicidin, LL-37, is small antimicro-
bial peptide, which reveals also some immunomodulatory
and proangiogenic properties and, therefore, may promote
wound healing. The expression of LL-37 is controlled by
various factors, including vitamin D. Thus, any distur-
bances in vitamin D level may influence LL-37 production
and, possibly, affect wound healing. Since deficiency of
vitamin D was identified as a common problem in the
population, this proof of concept study aimed to verify the
relationship between serum levels of LL-37, vitamin D,
and healing rate of venous leg ulcers. The study involved
small group (n = 19) of patients with venous leg ulcers.
Apart from non-venous ulcer aethiology, compression
intolerance, active vein thrombosis, and wound infection,
the exclusion criteria concerned also kidney insufficiency.
The results of the analysis of wound healing rates were
correlated with patients’ serum levels of 25(OH) vitamin D
and LL-37. In addition, serum levels of pro-inflammatory
cytokines (IL-6, IL-8, and TNF) were analyzed. We have
found strong association between serum concentrations of
LL-37 and the healing rates in patients with leg ulcers.
Despite the fact that 25(OH) vitamin D levels in all patients
were below the normal range, they did not show any cor-
relation with healing rates. Furthermore, no association
was observed between serum levels of 25(OH) vitamin D
and LL-37. No significant correlation between tested pro-
inflammatory cytokines and healing rate, LL-37, or 25(OH)
vitamin D levels was also observed. Regardless of small
study group, our results suggest that the assessment of
serum concentration of LL-37, but not 25-hydroxy vitamin
D, may help in predicting the wound healing efficacy.
Moreover, this assessment may be useful in pre-selection
of patients, which could benefit from local treatment with
exogenous LL-37.
Keywords Cathelicidin Healing rate LL-37
Venous leg ulcer Vitamin D
The Impact of Sex and Age on the Prevalence of Clinically
Relevant Sensitization and Asymptomatic Sensitization
in the General Population
Anna Dor-Wojnarowska1 • Jerzy Liebhart1 • Jadwiga Miecielica1 •
Marek Rabski1 • Andrzej Fal1 • Bolesław Samolin´ski2 • Marita Nittner-Marszalska1
Received: 20 January 2016 / Accepted: 4 July 2016 / Published online: 20 September 2016
Ó The Author(s) 2016. This article is published with open access at Springerlink.com
Abstract The objective of our study was to evaluate the
impact of sex and age on the prevalence of sensitization to
inhalant allergens. The study was performed as a part of
Polish Epidemiology of Allergic Diseases study, and data
concerning citizens of Wroclaw were analyzed. The par-
ticipants were divided into three age groups (6–7, 13–14,
and 20–44 years) with a subdivision according to sex. We
randomly selected 1409 individuals, 439 people complied;
the complete set of tests was performed on 421 of them.
We found that 37.7 % of the study population demon-
strated sensitization to at least one of the allergens tested.
Positive skin tests were found more frequently in males
than in females (p = 0.003); among 6–7-year-old children,
the sensitization was independent of sex (p = 0.26), while
in two other groups, it was higher in males (p = 0.002 and
p = 0.03, respectively). Clinically asymptomatic sensiti-
zation (AS) was found more often in females than in males
(p = 0.04). The higher rate of AS in women was observed
only in the two younger age groups, while in the 20–44-
year-old group AS did not differ between the sexes
(p = 0.72). Female sex hormones may contribute to a later
change in the nature of sensitization from clinically
asymptomatic to symptomatic. Further studies are needed
to confirm the results of our study.
Keywords Asymptomatic sensitization
Population study ‘‘Prick’’ skin test Total IgE
MAVS is not a Likely Susceptibility Locus for Addison’s Disease
and Type 1 Diabetes
Magdalena Zurawek1 • Marta Fichna1,2 • Marta Kazimierska1 • Piotr Fichna3 •
Agnieszka Dzikiewicz-Krawczyk1 • Grzegorz Przybylski1 • Marek Ruchala2 •
Jerzy Nowak1
Received: 2 March 2016 / Accepted: 31 August 2016 / Published online: 20 September 2016
Ó The Author(s) 2016. This article is published with open access at Springerlink.com
Abstract Mitochondrial antiviral signaling (MAVS) pro-
tein is an intracellular adaptor molecule, downstream of
viral sensors, retinoid acid-inducible gene I (RIG-I)-like
receptors (RLRs). Impaired antiviral cell signaling might
contribute to autoimmunity. Studies have recently shown
variations in genes encoding RLRs as risk factors for
autoimmune diseases. We investigated whether MAVS
coding polymorphisms are associated with Addison’s dis-
ease (AD) and type 1 diabetes (T1D) in Polish population.
We genotyped 140 AD, 532 T1D patients and 600 healthy
controls for MAVS rs17857295, rs7262903, rs45437096
and rs7269320. Genotyping was performed by TaqMan
assays. Distribution of the MAVS genotypes and alleles did
not reveal significant differences between patients and
controls (p [ 0.05). This analysis did not indicate the
association of the MAVS locus with susceptibility to AD
and T1D.
Keywords Addison’s disease Type 1 diabetes
MAVS Variants
CRISPR/Cas9 Immune System as a Tool for Genome Engineering
Magdalena Hryhorowicz1 • Daniel Lipin´ ski1 • Joanna Zeyland1 • Ryszard Słomski1,2
Received: 3 February 2016 / Accepted: 16 September 2016 / Published online: 3 October 2016
Ó The Author(s) 2016. This article is published with open access at Springerlink.com
Abstract CRISPR/Cas (clustered regularly interspaced
short palindromic repeats/CRISPR-associated) adaptive
immune systems constitute a bacterial defence against
invading nucleic acids derived from bacteriophages or
plasmids. This prokaryotic system was adapted in molec-
ular biology and became one of the most powerful and
versatile platforms for genome engineering. CRISPR/Cas9
is a simple and rapid tool which enables the efficient
modification of endogenous genes in various species and
cell types. Moreover, a modified version of the CRISPR/
Cas9 system with transcriptional repressors or activators
allows robust transcription repression or activation of tar-
get genes. The simplicity of CRISPR/Cas9 has resulted in
the widespread use of this technology in many fields,
including basic research, biotechnology and biomedicine.
Keywords CRISPR/Cas9 Genome editing
Off-target effect Guide RNA
Soluble BAFF Cytokine Levels and Antibody-Mediated Rejection
of the Kidney Allograft
Antonij Slavcev1 • Jitka Brozova1 • Janka Slatinska2 • Zuzana Sekerkova1 •
Eva Honsova3 • Jelena Skibova4 • Ilja Striz5 • Ondrej Viklicky2
Received: 9 June 2016 / Accepted: 2 November 2016 / Published online: 12 January 2017
Ó L. Hirszfeld Institute of Immunology and Experimental Therapy, Wroclaw, Poland 2016
Abstract The B-cell activating factor (BAFF) cytokine
has important functions for the survival and maturation of
B lymphocytes, which implies that this cytokine might play
a role in the development of antibody-mediated rejection
(AMR) after kidney transplantation. In our study, we
compared the concentrations of the soluble BAFF cytokine
in kidney graft recipients with AMR and patients without
rejection with the goal of testing the hypothesis whether
BAFF level measurement might be useful as a diagnostic
marker of AMR. The study included a cohort of 19 high-
risk patients with diagnosed AMR and 17 control patients
free of rejection. BAFF was measured in all patients before
transplantation, during the rejection episodes, and three
months after transplantation in patients free of rejection
using the Luminex technique. Before transplantation, the
serum concentrations of BAFF in patients with AMR and
kidney recipients without rejection did not significantly
differ. After transplantation, however, BAFF levels were
significantly lower in patients with AMR and also in
patients with concurrent humoral and cellular rejection
compared with patients without rejection (p \ 0.05 and
p \ 0.01, respectively). No correlation was found between
BAFF and the production of donor-specific antibodies
(DSA) before and after transplantation. Patients experi-
encing AMR and simultaneous cellular and AMR had
significantly lower concentrations of BAFF in comparison
with patients free of rejection.
Keywords BAFF Kidney transplantation
Antibody-mediated rejection HLA
The Frequency of CCR5del32 Mutation in Populations
of Russians, Tatars and Bashkirs of Chelyabinsk Region,
Russia
Irina Govorovskaya1,2 • Elena Khromova1 • Tatiana Suslova1,2 • Leonid Alexeev3 •
Ilya Kofiadi3
Received: 20 June 2016 / Accepted: 18 November 2016 / Published online: 12 January 2017
Ó L. Hirszfeld Institute of Immunology and Experimental Therapy, Wroclaw, Poland 2016
Abstract The distribution of genetic variants associated
with natural resistance to viral infections can vary among
human ethnic groups due to evolutionary factors, defining
the different epidemiologic background of world popula-
tions. The polymorphisms, defining the natural resistance
to HIV-infection and the rate of progression up to AIDS,
are very important since epidemic is still on rise. We have
studied the distribution of allele and genotype frequencies
of CCR5delta32 mutation in major populations inhabiting
Chelyabinsk region of the Russian Federation. Genetic
survey included the population of 509 potential blood
marrow donors: Russians (N = 300), Bashkirs (N = 118)
and Tatars (N = 91). The genotyping assay was performed
using real-time polymerase chain reaction (real-time PCR).
The genotypes were defined by melting curve analysis. The
CCR5delta32 allele and CCR5delta32/delta32 genotype
are presented in population of Russians in Chelyabinsk
region with the frequencies of Fx = 10.83% and
Px = 1.67, for the CCR5delta32 allele and its homozy-
gosity, respectively. In populations of Bashkirs and Tatars
CCR5delta32 allele and CCR5delta32/delta32 genotype
are presented at lower frequencies of Fx = 6.36%/
Px = 0.85 and Fx = 7.14%/Px = 1.10, respectively. These
data are consistent with the theory of northern origin of the
CCR5delta32 mutation.
Keywords HIV CCR5 Polymorphism
Stem cell donor registry
Intragenic Variations in BTLA Gene Influence mRNA Expression
of BTLA Gene in Chronic Lymphocytic Leukemia Patients
and Confer Susceptibility to Chronic Lymphocytic Leukemia
Lidia Karabon1,2 • Anna Partyka1 • Monika Jasek3 • Ewa Lech-Maranda4,5 •
Olga Grzybowska-Izydorczyk6 • Agnieszka Bojarska-Junak7 • Edyta Pawlak-Adamska1 •
Anna Tomkiewicz1 • Tadeusz Robak6 • Jacek Rolinski7 • Irena Frydecka1
Received: 21 June 2016 / Accepted: 18 November 2016 / Published online: 8 December 2016
Ó The Author(s) 2016. This article is published with open access at Springerlink.com
Abstract The aim of this study was to determine the
association between polymorphisms in gene encoding B-
and T-lymphocyte attenuator (BTLA) and susceptibility to
chronic lymphocytic leukemia (CLL) and their influence
on mRNA expression of BTLA gene in T and B cells from
CLL patients (pts.). The following BTLA single-nucleotide
polymorphisms (SNPs): rs2705511, rs1982809, rs9288952,
rs76844316, rs16859633, rs9288953, rs2705535,
rs1844089, rs2705565, rs2633580 were genotyped with use
of TaqMan probes in 321 CLL pts. and in 470 controls. The
mRNA levels of human BTLA were determined in sub-
populations of T and B cells from 37 CLL patients with use
of Applied Biosystems assays. Three SNPs: rs1982809,
rs2705511 and rs9288953 were associated with suscepti-
bility to CLL. The frequency of rs1982809[G] allele and
rs2705511[C] allele carriers was higher in patients com-
pared to the controls (0.51 vs. 0.41, OR 1.51, 95% CI
1.14–2.02, p = 0.004 and 0.56 vs. 0.44, OR 1.62, 95% CI
1.22–2.16, p = 0.0009, respectively). Furthermore,
rs9288953[TT] genotype was overrepresented in CLL pts.
compared to the controls (0.22 vs. 0.14, OR 1.74, 95% CI
1.20–2.53, p = 0.004). The evaluation of the influence of
BTLA SNPs on BTLA mRNA expression in CLL pts.
showed that the presence of rs1982809[G] allele was
associated with lower median (±SD) BTLA mRNA
expression in T cells (expressed as 2-delta Ct) in CLL pts.
as compared to [AA] homozygotes (0.009 ± 0.013 vs.
0.026 ± 0.012, p = 0.03). Our results indicate that
rs1982809 BTLA gene polymorphism is associated with
mRNA expression level and that variations in the BTLA
gene might be considered as potentially low-penetrating
CLL risk factor.
Keywords BTLA Gene polymorphisms
mRNA expression Chronic lymphocytic leukemia
Genetic Structure of the Armenian Population
Levon Yepiskoposyan1 • Anahit Hovhannisyan1 • Zaruhi Khachatryan1
Received: 24 June 2016 / Accepted: 20 October 2016 / Published online: 12 January 2017
Ó L. Hirszfeld Institute of Immunology and Experimental Therapy, Wroclaw, Poland 2016
Abstract Located at the crossroads of Europe and the
Middle East, the Armenian Highland served as a transition
corridor for major waves of prehistoric and historic
migrations. The genetic history of Armenians as an
indigenous population of the region attracts keen scientific
interest to resolve the puzzle of ancient Middle Eastern
populations’ expansion and the spread of Indo-European
languages. Here, we review the current state of studies on
the genetic structure of both modern and ancient inhabi-
tants of the Armenian Highland and outline further steps to
be fulfilled in this regard.
Keywords Armenian Highland Armenian population
Genetic structure
Genetic Factors of Diabetes
Karolina Antosik1 • Maciej Borowiec1
Received: 24 June 2016 / Accepted: 24 October 2016 / Published online: 12 January 2017
Ó L. Hirszfeld Institute of Immunology and Experimental Therapy, Wroclaw, Poland 2016
Abstract Monogenic diabetes is a rare genetic type of
diabetes caused by pancreatic b-cells dysfunction. All
subtypes of monogenic diabetes are recognized in the
pediatric population. They include maturity onset diabetes
of the young, permanent neonatal diabetes mellitus and
rare syndromic forms of diabetes. An early and proper
diagnosis allows to implement an optimal treatment, leads
to improved metabolic control and amelioration of related
disabilities as well as increases the quality of life of the
patients.
Keywords Monogenic diabetes MODY PNDM
The Expression of Toll-Like Receptors in Patients with B-Cell
Chronic Lymphocytic Leukemia
Justyna Rybka1 • Aleksandra Butrym2 • Tomasz Wro´bel1 • Bo_zena Jaz´wiec1 •
Aleksandra Bogucka-Fedorczuk1 • Rafał Pore˛ba3 • Kazimierz Kuliczkowski1
Received: 28 June 2016 / Accepted: 21 November 2016 / Published online: 12 January 2017
Ó The Author(s) 2016. This article is published with open access at Springerlink.com
Abstract B-cell chronic lymphocytic leukemia (B-CLL)
presents with progressive accumulation of monoclonal B
cells in the peripheral blood, bone marrow and lymphoid
organs. B-CLL is characterized by heterogeneous clinical
outcome. The expression of Toll-like receptors (TLRs) and
their association with other prognostic factors in B-CLL
patients remain unclear. The aim of our study was to
evaluate the expression of TLR2, TLR4 and TLR9 genes
and their significance as biological markers in patients with
B-CLL. Sixty patients with newly diagnosed B-CLL were
evaluated. The healthy control group included 20 age-
matched individuals. Using quantitative reverse transcrip-
tase PCR, the mRNA expression of genes TLR2, TLR4 and
TLR9 was measured. TLR4 gene expression was lower in
B-CLL patients as compared to the control group and
TLR2 gene expression was higher in B-CLL patients than
in healthy individuals. TLR9 gene expression was higher in
the control group than in patients with B-CLL. TLR4
mRNA expression was lower in patients with advanced-
stage CLL (Rai stages III and IV) than in patients with
early stage disease (Rai stages 0–II). TLR2 gene expression
was higher in patients with advanced-stage CLL (Rai
stages III and IV) than in patients with early stage disease
(Rai stages 0–II; p \ 0.05). Our results suggest that TLRs
could become potential biological markers for the clinical
outcome in patients with B-CLL.
Keywords Toll-like receptors
Chronic lymphocytic leukemia
An Association Between Functional Polymorphisms
of the Interleukin 1 Gene Complex and Schizophrenia Using
Transmission Disequilibrium Test
Pawel Kapelski1 • Maria Skibinska1 • Malgorzata Maciukiewicz2 •
Joanna Pawlak1 • Monika Dmitrzak-Weglarz1 • Aleksandra Szczepankiewicz3 •
Dorota Zaremba1 • Joanna Twarowska-Hauser1
Received: 21 July 2016 / Accepted: 28 October 2016 / Published online: 12 January 2017
Ó L. Hirszfeld Institute of Immunology and Experimental Therapy, Wroclaw, Poland 2016
Abstract IL1 gene complex has been implicated in the
etiology of schizophrenia. To assess whether IL1 gene
complex is associated with susceptibility to schizophrenia
in Polish population we conducted family-based study.
Functional polymorphisms from IL1A (rs1800587,
rs17561, rs11677416), IL1B (rs1143634, rs1143643,
rs16944, rs4848306, rs1143623, rs1143633, rs1143627)
and IL1RN (rs419598, rs315952, rs9005, rs4251961) genes
were genotyped in 143 trio with schizophrenia. Statistical
analysis was performed using transmission disequilibrium
test. We have found a trend toward an association of
rs1143627, rs16944, rs1143623 in IL1B gene with the risk
of schizophrenia. Our results show a protective effect of
allele T of rs4251961 in IL1RN against schizophrenia. We
also performed haplotype analysis of IL1 gene complex
and found a trend toward an association with schizophrenia
of GAGG haplotype (rs1143627, rs16944, rs1143623,
rs4848306) in IL1B gene, haplotypes: TG (rs315952,
rs9005) and TT (rs4251961, rs419598) in IL1RN. Haplo-
type CT (rs4251961, rs419598) in IL1RN was found to be
associated with schizophrenia. After correction for multiple
testing associations did not reach significance level. Our
results might support theory that polymorphisms of
interleukin 1 complex genes (rs1143627, rs16944,
rs1143623, rs4848306 in IL1B gene and rs4251961,
rs419598, rs315952, rs9005 in IL1RN gene) are involved
in the pathogenesis of schizophrenia, however, none of the
results reach significance level after correction for multiple
testing.
Keywords Genetics Polymorphism Schizophrenia
Transmission disequilibrium test
Interleukin 1 gene complex
Distribution of HLA-DQB1 in Czech Patients with Central
Hypersomnias
Milena Vrana1 • Vera Siffnerova1 • Pavla Pecherkova1 • Eva Ratajova1 •
Karel Sonka1,2
Received: 1 August 2016 / Accepted: 18 November 2016 / Published online: 12 January 2017
Ó L. Hirszfeld Institute of Immunology and Experimental Therapy, Wroclaw, Poland 2016
Abstract The aim of our study was to analyze the distri-
bution of HLA-DQB1 in Czech patients with central
hypersomnias and differences between diagnostic groups
of narcolepsy type 1 (NT1), type 2 (NT2), idiopathic
hypersomnia (IH) and no central hypersomnia subjects (no-
CH). Statistical analysis of DQB1 genotyping was per-
formed on the cohort of 716 patients (375 men, 341
women) with reported excessive daytime sleepiness.
DQB1*06:02 allele was present in 94% of the NT1
patients. The decrease of DQB1*06:03 allele was also
confirmed. No other DQB1*06 allele nor any other DQB1
allele group was differently distributed in the NT1. In the
cohort of NT2 patients DQB1*06:02 allele was present in
43%. Allele group DQB*05 was detected with a signifi-
cantly higher frequency in this diagnostic unit. Any
differences in presence of DQB1*05 alleles in NT2
patients were not reported so far. The cohort of patients
with IH did not show any difference in allele distribution of
DQB1 alleles/allele groups comparing to healthy controls.
DQB1*06:02 allele was significantly increased in the no
hypersomnia group. No other DQB1 allele/allele group had
any difference in distribution in patients comparing to
healthy controls. The different distribution of DQB1*06:02
and other DQB1 alleles/allele groups was detected in
analyzed diagnostic groups. These results indicate that
DQB1 contributes to the genetic predisposition to NT1,
NT2, IH and no-CH in different manners.
Keywords Narcolepsy Idiopathic hypersomnia
Central hypersomnias HLA-DQB1
Single Nucleotide Polymorphisms of the ERAP1 Gene and Risk
of NSCLC: A Comparison of Genetically Distant Populations,
Chinese and Caucasian
Yufeng Yao1 • Andrzej Wis´niewski2 • Qiangli Ma3 • Aneta Kowal4 •
Irena Pore˛bska4 • Konrad Pawełczyk5 • Jiankun Yu1 • Joanna Dubis6 •
Natalia _Zuk6 • Yingfu Li7 • Li Shi1 • Piotr Kus´nierczyk2
Received: 6 June 2016 / Accepted: 18 October 2016 / Published online: 12 January 2017
Ó The Author(s) 2016. This article is published with open access at Springerlink.com
Abstract An effective cytotoxic immune response to
neoplastic cells requires efficient presentation of antigenic
peptides to T lymphocytes by HLA class I (HLA-I)
molecules. The HLA-I-bound peptide repertoire depends
on antigen-processing machinery molecules. Aminopepti-
dase residing in endoplasmic reticulum 1 (ERAP1) trims
peptides to the optimal length for HLA-I binding. Single
nucleotide polymorphisms (SNPs) in the ERAP1 gene
result in changes in aminopeptidase activity and specificity.
This may affect susceptibility to cancer. However, non-
small cell lung carcinoma (NSCLC) has not been studied in
this respect. We compared genotype and haplotype fre-
quencies of four coding, nonsynonymous ERAP1 SNPs,
rs26653G [ C, rs26618T [ C, rs30187C [ T, and
rs27044C [ G, in NSCLC occurring in two genetically
distant populations, Chinese and Poles. We found associ-
ations of all four SNPs with NSCLC in Chinese but not in
Poles. The differences in ERAP1-NSCLC associations
might be explained by highly significant differences in
SNP genotype frequencies between Chinese and Poles
(except for rs26618). In accordance with this, the most
frequent ERAP1 haplotypes were distributed differently in
cases versus controls in Chinese, but not in Poles. Our
findings add to the differences between Orientals and
Caucasians in genetics of disease susceptibility.
Keywords Non-small cell lung carcinoma
Endoplasmic reticulum aminopeptidase-1
Genetic association
The Influence of HLA and KIR Genes on Malignant Melanoma
Development and Progression
Snezhina Mihailova Kandilarova1 • Annette Paschen2 • Anastassia Mihaylova1 •
Milena Ivanova1 • Dirk Schadendorf2 • Elissaveta Naumova1
Received: 13 June 2016 / Accepted: 25 October 2016 / Published online: 12 January 2017
Ó L. Hirszfeld Institute of Immunology and Experimental Therapy, Wroclaw, Poland 2016
Abstract Many studies have described the role of killer
immunoglobulin-like receptors (KIRs) and their cognate
human leukocyte antigen (HLA) class I ligands in the
immune protection against melanoma, but the effect of
these markers on intra-individual variations in tumor
development and progression has remained less clear. We
performed KIR, HLA, and KIR/ligand analysis in 283
patients with malignant melanoma in order to evaluate their
integrated influence on disease stage and progression. The
patients were grouped according to AJCC staging, histo-
logical type of the primary tumor, progression, and survival
rate. Analysis of HLA class I alleles revealed positive
association of HLA-C*14 (Pc = 0.026, OR = 5.99) and
negative association of HLA-C*02 (Pc = 0.026,
OR = 0.43) with the disease. Decreased frequency of
KIR2DS5 was observed in patients with rapid progression,
as compared to those with slow progression. KIR BB
genotype was prevalent in patients with metastasis
(p = 0.004, OR = 0.025). KIR AA genotype was nearly
twice as frequent in rapidly progressive cases, but without
statistical relevance (p = 0.055, OR = 2.6). Significantly
increased frequency of KIR2DL2 in the presence of C1
ligand (strong inhibition) was found in patients with AJCC
III and IV, as compared to individuals with AJCC I stage
(p = 0.045, OR = 1.93). In summary, our data imply that
KIR/ligand gene content in patients could modulate the
disease course towards unfavorable tumor behavior.
Keywords Malignant melanoma Progression
Inhibitory killer immunoglobulin-like receptors (KIR)
Human leukocyte antigens (HLA)
Prediction of NK Cell Licensing Level in Selection
of Hematopoietic Stem Cell Donor, Initial Results
Marta Rogatko-Koros´1 • Renata Mika-Witkowska1 • Katarzyna Bogunia-Kubik2 •
Barbara Wysoczan´ ska2 • Emilia Jaskuła2,3 • Katarzyna Kos´cin´ ska3 •
Klaudia Nestorowicz1 • Joanna Dziopa1 • Urszula Szlendak1 • Sławomir Gwozdowicz1 •
El_zbieta Graczyk-Pol1 • Andrzej Lange2,3 • Jacek Nowak1
Received: 5 July 2016 / Accepted: 18 November 2016 / Published online: 8 December 2016
Ó L. Hirszfeld Institute of Immunology and Experimental Therapy, Wroclaw, Poland 2016
Abstract Natural killer (NK) cell licensing status depends
on clonal expression of inhibitory killer cell
immunoglobulin-like receptors (iKIR) and short term HLA
environment. Licensed NK cells are more efficient in tumor
killing than unlicensed NK cells. Cognate KIR–HLA pairs
in hematopoietic stem cell transplant (HSCT) donor and
recipient are decisive for the possible change in the NK cell
licensing status after HSCT. We assessed clinical outcomes
in 297 patients with lymphoproliferative or myeloprolif-
erative malignancies, or myelodysplastic syndrome in a
model with upward licensing, downward resetting, and
unchanged licensing genetics status after T cell replate
HSCT from unrelated donors. We found extremely low
(0%) relapse/progression incidence (RI), and better (59%)
event-free survival (EFS) in recipients with upward
licensing status and highly increased RI (37.5%), and
reduced EFS (8%) among patients with the downward
resetting status of repopulated donor NK cells after HSCT,
as compared with unchanged NK cell licensing (RI 23%,
EFS 47%). These trends were confirmed in adjusted mul-
tivariable models (for RI p = 6.66E–09, OR = 1.47, 95%
CI 1.29–1.66 and for EFS p = 3.79E–13, OR = 1.67,
95% CI 1.50–1.84). Differences in the incidence of acute
graft versus host disease (GvHD 62, 69, and 47%) and
chronic GvHD (24, 44, and 15%, respectively) in three
groups were insignificant. It would be rationale the pref-
erential selection of the donors with upward licensing over
downward resetting inhibitory KIR:HLA constellation and
inclusion of the KIR genotyping in the donor selection
algorithm for malignant patients. Further studies using
enlarged cohorts of patients with more homogenous diag-
nosis are essential to reliably verify these preliminary data.
Keywords NK cell licensing
Hematopoietic stem cell transplantation
Unrelated donors Bone marrow donor selection
Hematopoietic malignancy Relapse
Transplantology: Challenges for Today
Maria Boratyn´ ska1 • Dariusz Patrzałek2
Received: 29 July 2016 / Accepted: 20 October 2016 / Published online: 12 January 2017
Ó The Author(s) 2016. This article is published with open access at Springerlink.com
Abstract Clinical transplantology in Poland had its 50th
anniversary this year. With the early and long results
comparable to the best achieved in the world leading
centers, we face old and completely new challenges for this
medical speciality. Main and growing challenge is insuf-
ficient number of available organs. With less than
15 donors/mln population/year Poland stay in the lower
row of European countries in this measurement of trans-
plant activity. Donation system is not efficient enough and
we lose a big number of potential donors still. Living
donation (with the exception for the fragments of the liver)
remains low despite of different initiatives made so far on
the national and local levels. Donation after cardiac death
is possible from the point of Polish juridical regulations,
but since last 3 years had not showed real impact on
country donation rates (only three procedures done).
Methods of tissue typing remain slow and cause relatively
long times of cold ischemia for kidney programs. Second
main challenge is chronic rejection causing loss of organs
in the long-term follow-up and no efficient treatment
employed. The emerging possibility of tolerance induction
despite of plenty of new protocols proposition in the pub-
lications does not show up a clinical everyday practice in
work. The same is with xenotransplantation promises; even
we were informed recently that till 2030 such genetically
modified porcine organs will be available. The next chal-
lenge is production of organs and tissues from own
recipients cells installed on the different scaffolds or 3D
printed. Other challenge is the personnel working in this
field. We observe like in the other European countries lack
of new candidates for work in this field together with
serious problems of nursing staff, being a catastrophic
perspective in country medical service in general, not only
in transplant centers. The last but not least challenge is
financial side of transplant programs.
Keywords Clinical transplantation Organ donation
Chronic rejection Tolerance
Characterization of Three CYP2C19 Gene Variants
by MassARRAY and Point of Care Techniques: Experience
from a Czech Centre
Martin Petrek1,2,3 • Lenka Kocourkova1,3 • Veronika Zizkova1,3 • Zdenek Nosek3 •
Milos Taborsky4 • Jana Petrkova3,4
Received: 29 July 2016 / Accepted: 20 October 2016 / Published online: 12 January 2017
Ó L. Hirszfeld Institute of Immunology and Experimental Therapy, Wroclaw, Poland 2016
Abstract Distribution of cytochrome P450 2C19 enzyme
gene (CYP2C19) variants affecting metabolism of clopi-
dogrel was determined in 526 Czech patients after
percutaneous coronary intervention using MassARRAY
genotyping and compared to distribution in other popula-
tions of European descent. Fifty-three (10%) patients
underwent parallel determination of CYP2C19 genotypes
from buccal swabs by a point of care technique with 100%
concordance to the main genotyping platform. Observed
CYP2C19 genotypes were related to clopidogrel metabo-
lism phenotypes and discussed in population context.
Hereby, presented methodologies provide accurate
CYP2C19 genotyping results in a relatively short time of
one up to 12 h and may, therefore, find the relevant place in
the field of genotype-guided antiplatelet therapy.
Keywords CYP2C19 Pharmacogenetics Clopidogrel
Genotyping Point of care
Functional Polymorphism in the Interleukin 6 (IL6) Gene
with Respect to Depression Induced in the Course of Interferon-a
and Ribavirin Treatment in Chronic Hepatitis Patients
Dorota Frydecka1 • Tomasz Pawłowski1 • Dariusz Pawlak2 • Krzysztof Małyszczak1,3
Received: 13 September 2016 / Accepted: 18 November 2016 / Published online: 12 January 2017
Ó The Author(s) 2016. This article is published with open access at Springerlink.com
Abstract Interleukin (IL)-6 is a multifactorial cytokine
known to be increased in patients with chronic hepatitis C
(CHC) and to be predictive of depression incidence. The
aim of the study was to explore the association between IL6
gene C-174G polymorphism and depressive symptom
severity in the longitudinal study design following the
course of pegylated interferon/ribavirin treatment in CHC
patients. In our study, we included 62 CHC subjects. They
were assessed using present state examination, Beck
Depression Inventory (BDI) and Montgomery A˚ sberg
Depression Rating Scale (MADRS) at weeks 0, 3, 5, 9, 13,
24 and 24 weeks after the end of treatment. The risk of
depression was associated with higher baseline MADRS
score and BDI score. Interestingly, when stratified by IL6
C-174G polymorphism, higher baseline depressive symp-
tom severity measured by MADRS and BDI predicted
higher risk of depression in the course of antiviral treat-
ment only in high IL-6 producers—G allele carriers
(patients with GG and CG genotypes) (p = 0.004,
p = 0.00008, respectively). There is interaction between
severity of baseline depressive symptoms at the beginning
of antiviral therapy and IL6 gene C-174G polymorphism
leading to increased risk for the development of depressive
episode in CHC patients in the course of antiviral
treatment.
Keywords Chronic hepatitis C Depression
Interleukin-6 Functional genetic polymorphism
Longitudinal study Hepatitis C virus
Cereblon and IRF4 Variants Affect Risk and Response
to Treatment in Multiple Myeloma
Aleksandra Butrym1 • Piotr Łacina2 • Justyna Rybka3 •
Monika Chaszczewska-Markowska2 • Grzegorz Mazur4 •
Katarzyna Bogunia-Kubik2,4
Received: 29 June 2016 / Accepted: 24 October 2016 / Published online: 12 January 2017
Ó The Author(s) 2016. This article is published with open access at Springerlink.com
Abstract Multiple myeloma (MM) is a plasma-cell
malignancy derived from an early precursor of the B-cell
lineage characterised by bone-marrow infiltration, lytic
bone lesions, and the presence of a monoclonal protein in
serum and/or urine. Interferon regulatory factor 4 (IRF4) is
a critical transcriptional regulator in B-cell development
and function that is required during immune response for
lymphocyte activation and the generation of
immunoglobulin-secreting plasma cells. Immunomodula-
tory drugs, derivatives of thalidomide, are commonly used
in therapy against MM. They are known to target a protein
called cereblon (CRBN); however, the exact mechanism
remains unknown. The present study aimed to assess the
association of two (rs12203592 and rs872071) polymor-
phisms within the IRF4 gene and two (rs711613 and
rs1045433) in the CRBN gene with MM susceptibility,
progression, and response to treatment. For this purpose,
144 MM patients and 126 healthy individuals were geno-
typed for the IRF4 and CRBN alleles. The presence of the
IRF4 (rs872071) G allele was more frequently detected in
patients than healthy individuals (OR 1.78; P = 0.034),
and this relationship was especially pronounced in women
(OR 2.83; P = 0.012). The CRBN (rs711613) A allele-
carriers were better responders to the treatment
(P = 0.012), in particular to thalidomide including therapy
(P = 0.023). These results underline the prognostic sig-
nificance of the IRF4 and CRBN polymorphisms in patients
with MM.
Keywords Cereblon Interferon regulatory factor 4
Single nucleotide polymorphisms Multiple myeloma
Disease susceptibility Stage of the disease
Response to treatment
Significance of Polymorphism and Expression of miR-146a
and NFkB1 Genetic Variants in Patients with Rheumatoid
Arthritis
Katarzyna Bogunia-Kubik1,2 • Barbara Wysoczan´ ska1 • Dagmara Pia˛tek1 •
Milena Iwaszko1 • Marzena Ciechomska1,4 • Jerzy S´ wierkot3
Received: 21 July 2016 / Accepted: 18 October 2016 / Published online: 12 January 2017
Ó The Author(s) 2016. This article is published with open access at Springerlink.com
Abstract MicroRNA-146a (miR-146a) has been shown to
play an important role in the regulation of inflammatory
innate immune responses, and found to be differentially
expressed in rheumatoid arthritis (RA). Through NF-jB
pathway, this molecule is able to stimulate the release of pro-
inflammatory cytokines such as TNF-a, IL-1b, and IL-17. It
has been also suggested that single-nucleotide polymor-
phisms (SNPs) in miRNA sequences may alter miRNA
expression and that miR-146a rs2910164 SNP may con-
tribute to RA development. These observations prompted us
to analyze the potential associations between the miR–146a–
3p (rs2910164, G [ C) and NFkB1 (rs28362491, ins/del
ATTG) polymorphisms and miR-146a-5p expression in
patients’ sera in relation to clinical outcome of the treatment
as well as predisposition to RA. Genotyping was performed
in 111 patients and 130 healthy individuals while 16 controls
and 13 RA patients (before and after three months of therapy
with TNF-a inhibitors (TNFi)) were studied for the circu-
lating miR-146a-5p serum expression level. Patients
carrying the NFkB1 ins/ins genotype were characterized by
worse response to TNFi treatment (p = 0.023). In patients,
before TNFi therapy, expression levels of miR-146a-5p were
less (0.422 ± 0.171) as compared to those detected after
three months of treatment (1.809 ± 0.658, p = 0.033) and
observed for healthy controls (5.302 ± 2.112, p = 0.048).
Moreover, patients with higher circulating miR-146a-5p
levels after three months of TNFi administration were more
frequently carrying the rs2910164-C allele (p = 0.032).
These results support the hypothesis that miR-146a might be
involved in pathogenesis of RA and imply that miR–146a–3p
polymorphism may be associated with miR-146a-5p levels
in serum after anti-TNF-a treatment.
Keywords Rheumatoid arthritis
miRNA–146a–3p polymorphism
miRNA-146a-5p serum level NFkB1 polymorphism
Disease susceptibility Response to treatment
ERAP1 and ERAP2 Gene Variations Influence the Risk
of Psoriatic Arthritis in Romanian Population
Olivia M. Popa1 • Marius Cherciu1 • Laura I. Cherciu1 • Monica I. Dutescu2 •
Mihai Bojinca3 • Violeta Bojinca4 • Constantin Bara1 • Luis O. Popa5
Received: 24 June 2016 / Accepted: 1 December 2016 / Published online: 12 January 2017
Ó L. Hirszfeld Institute of Immunology and Experimental Therapy, Wroclaw, Poland 2016
Abstract Psoriatic arthritis (PsA) is a chronic inflamma-
tory disorder that belongs to the group of spondyloarthritis
(SpA). It was found that single nucleotide polymorphisms
(SNPs) of endoplasmic reticulum aminopeptidase (ERAP1
and ERAP2) genes influence the risk of ankylosing
spondylitis, the most common form of SpA and the risk of
psoriasis. Our purpose was to investigate the possible
association of ERAP1 and ERAP2 gene SNPs with psori-
atic arthritis susceptibility in Romanian population.
Subsequent analyses included patients’ subgroups accord-
ing to HLA-B27 status. Psoriatic arthritis patients (N = 98)
and random healthy controls (N = 139) were genotyped
for ERAP1/2 genes SNPs rs30187, rs27044, rs2910686,
and rs2248374 by TaqMan Allelic Discrimination Assays.
An additional control group (N = 108; 100% HLA-B27
positive) was used for subsequent analyses. The results
showed the association of rs2248374 SNP of ERAP2 gene
with the risk of PsA, especially for HLA-B27 negative
disease (p = 0.02; OR 1.59). ERAP2 haplotype GT
(rs2248374/rs2910686) was significantly under-represented
in PsA patients than in controls (43 vs. 55%; p = 0.02).
The analysis of ERAP1 SNPs in HLA-B27 positive con-
trols and PsA subgroup showed strong evidence of
association for rs30187 (p = 0.005; OR 2.73) and for CC
rs30187/rs27044 haplotype (47% in patients vs. 70.5% in
controls; p = 0.006). In conclusion, we found a significant
association of ERAP2 with PsA and HLA-B27 negative
PsA, while ERAP1 association was restricted only to HLA-
B27 positive disease. To our knowledge, this is the first
study that investigated ERAP2 polymorphisms in relation
to PsA susceptibility.
Keywords Psoriatic arthritis ERAP1 ERAP2
Single nucleotide polymorphism Haplotype
Determination of HLA-A, -B, and -DRB1 Allele and Haplotype
Frequencies in the Croatian Population Based on a Family Study
Zorana Grubic1 • Marija Maskalan1 • Danijela Svilicic1 •
Katarina Stingl Jankovic1 • Renata Zunec1
Received: 29 June 2016 / Accepted: 1 December 2016 / Published online: 12 January 2017
Ó L. Hirszfeld Institute of Immunology and Experimental Therapy, Wroclaw, Poland 2016
Abstract In the present study, HLA allele and haplotype
frequencies among Croatian families were investigated to
evaluate valuable information about HLA genotypes and to
compare them with data from the Croatian Bone Marrow
Donor Registry (CBMDR). A total of 350 families which
have been typed for the purpose of HSCT were included.
All individuals were tested using PCR-SSO or PCR-SSP
methods for HLA-A, -B, and -DRB1 alleles. The HLA-A-
B-DRB1 haplotypes were determined by segregation and
directly counted. A total of 30 HLA-A, 54 HLA-B, and 38
HLA-DRB1 alleles and 716 different HLA-A-B-DRB1
haplotypes were identified. Of these, the three most fre-
quent alleles at HLA-A, -B, and -DRB1 loci, respectively,
were A*02:01 (30.39%), A*11:01 (13.37%), A*24:02
(10.91%); B*51:01 (12.48%), B*18:01 (8.35%), B*08:01
(8.06%); DRB1*03:01 (11.20%), DRB1*01:01 (9.84%),
DRB1*16:01 (9.63%). The following HLA alleles were
detected only once: A*02:09, A*24:03, A*24:04, A*24:07;
B*07:04, B*15:07, B*15:08, B*39:04, B*39:10, B*39:24,
B*40:04; DRB1*08:03, DRB1*11:06, DRB1*13:32,
DRB1*14:05. Five most frequent haplotypes were:
A*01:01-B*08:01-DRB1*03:01 (5.34%), A*02:01-B*
18:01-DRB1*11:04 (1.57%), A*02:01-B*27:02-DRB1*
16:01 (1.50%), A*02:01-B*27:05-DRB1*01:01 (1.42%)
and A*02:01-B*44:02:01G-DRB1*16:01 (1.28%). The
haplotype frequencies based on the family study were
compared with the frequencies from CBMDR, and similar
results were obtained for all except for the HLA-A*26:01-
B*38:01-DRB1*04:02 haplotype. A significantly higher
frequency (P = 0.0017) of this haplotype was observed
among family individuals. Nine haplotypes were unique
and data about their frequencies do not exist in current
databases. The data obtained in this study could be useful
for anthropology, transplantation and disease association
studies.
Keywords HLA Allele and haplotype frequencies
Family study Croatian population
Profile of Inflammation-Associated Proteins in Early Post-
Transplant Samples of Patients After Allogeneic Hematopoietic
Stem Cell Transplantation: a Preliminary Study
Frantisek Mrazek1 • Petra Schneiderova1 • Eva Kriegova1 • Ludek Raida2 •
Adam Kuba2 • Petr Gajdos3 • Nikola Ko¨nigova1 • Jana Onderkova1 •
Zuzana Ambruzova1
Received: 29 July 2016 / Accepted: 1 December 2016 / Published online: 12 January 2017
Ó L. Hirszfeld Institute of Immunology and Experimental Therapy, Wroclaw, Poland 2016
Abstract Allogeneic hematopoietic stem cell transplan-
tation (aHSCT) is used as a curative treatment in severe
hematological and immunological disorders. Despite clear
improvement of the aHSCT outcome, substantial pro-
portion of patients still suffers from severe complications,
including graft-versus-host disease (GvHD). The aim of
this study was, therefore, to identify inflammation-asso-
ciated molecules deregulated in the early serum samples
of the patients after aHSCT and nominate markers asso-
ciated with particular aHSCT parameters/complications.
Serum concentrations of 92 inflammation-associated
proteins were measured in samples obtained from 80
aHSCT patients 14 days after transplantation and from 23
healthy control subjects by a novel sensitive proximity
extension assay technology using Proseek Multiplex
Inflammation I kit. Serum profiles of inflammatory pro-
teins in patients after aHSCT were substantially different
from those observed in control subjects and related to
underlying disease status before transplantation. Particu-
larly, the difference between aHSCT patients and controls
reached significance level for 57 analytes (40 upregulated,
17 downregulated in aHSCT patients). The concentration
of several markers was associated with the level of donor/
recipient HLA match (TGF-a: pcorr = 0.025, HGF:
pcorr = 0.036) and with complete donor chimerism at day
?30 after allografting (DNER: pcorr = 0.042). None of
the markers was significantly associated with acute and
chronic GvHD after correction. More than half of inves-
tigated proteins significantly differed between the samples
from aHSCT patients and healthy control subjects as a
consequence of the ‘‘cytokine storm’’ after aHSCT.
Comparisons of patient’s subgroups based on specific
biological/clinical parameters revealed much less evident
differences; nevertheless, we nominated several markers
associated with the level of donor/recipient HLA match
and post-transplant chimerism.
Keywords Hematopoetic stem cell transplantation
Graft-versus-host disease Protein profiles Biomarkers
Inflammation
Mutation c.256_257delAA in RAG1 Gene in Polish Children
with Severe Combined Immunodeficiency: Diversity of Clinical
Manifestations
Anna Szaflarska1,2 • Magdalena Rutkowska-Zapała1,2 • Monika Kotula2 •
Anna Gruca1 • Agnieszka Grabowska3,4 • Marzena Lenart1,2 • Marta Surman2 •
El_zbieta Trzyna5 • Anna Mordel5,6 • Anna Pituch-Noworolska1,2 • Maciej Siedlar1,2
Received: 2 August 2016 / Accepted: 2 December 2016 / Published online: 12 January 2017
Ó The Author(s) 2016. This article is published with open access at Springerlink.com
Abstract Mutations in RAG1 gene may result in different
types of severe combined immunodeficiencies. In this
study, we compare clinical symptoms and laboratory
findings in four children with identical mutation in RAG1
gene. All of analyzed patients presented symptoms of
severe combined immunodeficiencies associated or not
with Omenn syndrome (OS) features. In our patients two
different types of variants in RAG1 gene were detected.
The first of the mutation was the deletion of AA dinu-
cleotide at position c.256_257 (p.Lys86ValfsTer33), the
second gene variant was substitution c.2867T[C
(p.Ile956Thr). In Patient 1 we detected that compound
heterozygous mutations involved both of the mentioned
variants. Whereas, in Patients 2, 3 and 4, we confirmed the
presence of the dinucleotide deletion but in a homozygous
state. In all described patients, sequence analysis of RAG2
gene did not reveal any nucleotide changes. Our data show
that mutation c.256_257delAA in RAG1 gene seems to
occur quite frequently in the polish patients with severe
combined immunodeficiency and may result in classical
OS as well as in severe combined immunodeficiency
without clinical and laboratory features of OS when
occurred in homozygous state. The same mutation but in
heterozygous state, in combination with other mutation in
RAG1 gene, may result in incomplete OS.
Keywords RAG1/2 genes Severe combined
immunodeficiency Omenn syndrome
Primary Biliary Cholangitis Associated with Skin Disorders:
A Case Report and Review of the Literature
Benedetta Terziroli Beretta‑Piccoli1 · Caroline Guillod2 · Igor Marsteller3 ·
Roland Blum4 · Luca Mazzucchelli5 · Chiara Mondino2 · Pietro Invernizzi6 ·
M. Eric Gershwin7 · Carlo Mainetti2
Abstract Primary biliary cholangitis (PBC) is a rare auto-
immune cholestatic liver disease. It is often associated with
extrahepatic autoimmune diseases. Skin disorders are spo-
radically reported in association with PBC. We report an
unusual case of PBC associated with acquired reactive per-
forating dermatosis (ARPD) and present a review of the lit-
erature on skin disorders associated with PBC. Our patient
presented to the dermatology department with generalized
pruritus associated with nodular perforating skin lesions on
the trunk, and cholestatic liver disease of unknown origin.
After having established both diagnosis of ARPD and PBC,
she was managed in an interdisciplinary manner, and both
her skin and liver conditions improved gradually. Only one
similar case is reported in the literature, in that case, the
liver disease was not treated. By reviewing the literature,
we found that lichen planus, vitiligo, and psoriasis are the
most frequent skin disorders associated with PBC. How-
ever, there is only limited data about specific skin disorders
associated with PBC. This case report of a patient with
PBC associated with ARPD underlines the importance of
interdisciplinary management of patients with rare liver
diseases combined with rare skin disorders. The present
review of the literature shows that probably, immune-medi-
ated skin conditions are not more frequent in PBC patients
than in the general population. However, the available data
are scant; there is a need for high-quality data on skin con-
ditions associated with PBC.
Keywords Primary biliary cholangitis · Acquired
reactive perforating dermatosis · Skin disorders · Case
report · Review of the literature
Abstracts
Proceedings of the 10th Jubilee East–West Immunogenetics
Conference on April 21–23, 2016 (Wrocław, Poland)
The Effect of Nonsurgical Periodontal Therapy on HNP1-3 Level
in Gingival Crevicular Fluid of Chronic Periodontitis Patients
Ewa Dolińska1 · Anna Skurska1 · Małgorzata Pietruska1,2 ·
Violetta Dymicka-Piekarska3 · Robert Milewski4 · Jan Pietruski2 · Anton Sculean5
Abstract The rich bacterial flora of oral cavity is con-
trolled by innate immune response, including antibacterial
peptides and among them human neutrophil peptides 1–3
(HNP1-3). The knowledge of the involvement of HNPs in
innate and acquired immunity of the periodontium is frag-
mentary. The aim of the study was to assess alterations in
HNP1-3 levels in the gingival crevicular fluid (GCF) of
chronic periodontitis patients before and after nonsurgi-
cal periodontal therapy. Nineteen patients with chronic
periodontitis were qualified to the study. After periodontal
examination, one site with pocket depth (PD) ≥4 mm was
selected. All the patients received periodontal treatment
involving scaling and root planing with additional sys-
temic antibiotic therapy (Amoxicillin 375 mg three times
daily and Metronidazole 250 mg three times daily for 7
days). Prior to therapy, 3 and 6 months after it, clinical
periodontal parameters were measured and GCF was col-
lected from previously chosen site. The level of HNP1-3 in
GCF was determined by means of a commercially available
enzyme-linked immunoassay kit. The periodontal therapy
caused a statistically significant (p < 0.001) decrease in
all the assessed clinical parameters at the sites of sample
collection except for bleeding on probing. The level of
HNP1-3 per measure point showed a statistically signifi-
cant increase (baseline—3 months: p = 0.05, baseline—6
months: p = 0.007). Within the limits of the study, it can be
stated that nonsurgical periodontal therapy with additional
systemic administration of Amoxicillin and Metronidazole
increases the level of HNP1-3 in GCF.
Keywords Chronic periodontitis · Gingival crevicular
fluid · Alpha-defensins · Human neutrophil peptide
Apportioning Blame: Autoreactive CD4+ and CD8+ T Cells
in Type 1 Diabetes
Rubén Varela‑Calvino1 · Cristina Calviño‑Sampedro1 · Iria Gómez‑Touriño2 ·
Oscar J. Cordero
Abstract Type 1 diabetes (T1D) is one of the most
studied archetypal organ-specific autoimmune diseases.
Although many clinical, epidemiological, and pathological
characteristics have been described, there are still impor-
tant issues which need to be resolved as these will have a
major impact on the development of future antigen-specific
immunotherapies. An important question relates to T lym-
phocytes in the development of the disease, in particular
their role in the destruction of insulin-producing beta cells.
Since the discovery that certain class II histocompatibility
complex molecules (HLA) are linked to the development
of T1D, much research has focused on CD4+ helper T lym-
phocytes; however, recent studies highlight class I HLA
molecules as an independent risk factor; hence, research
into the role played by CD8+ cytotoxic T lymphocytes has
gained momentum. In this review, we summarize recent
studies clarifying the role played by both sets of autoreac-
tive T lymphocytes in T1D, discuss the targets recognized
by these cells and their phenotype in T1D patients. Finally,
we will examine the possible generation of regulatory
CD8+ T lymphocytes upon different immuno-intervention
strategies.
Keywords Type 1 diabetes · T lymphocytes ·
Autoantigens · CD8 · Regulatory T cells
Functions of Cancer-Derived Extracellular Vesicles
in Immunosuppression
Liliana Czernek1 · Markus Düchler
Abstract Extracellular vesicles, including exosomes,
constitute an important element of intercellular communi-
cation by carrying a variety of molecules from producer
to target cells. The transport of mRNA and miRNA can
directly modulate gene expression in the target cells. The
miRNA content in exosomes is characteristic for the cell
from which the vesicles were derived enabling the usage of
exosomes as biomarkers for the diagnosis various diseases,
including cancer. Cancer-derived exosomes support the
survival and progression of tumors in many ways and also
contribute to the neutralization of the anti-cancer immune
response. Exosomes participate in all known mechanisms
by which cancer evades the immune system. They influ-
ence the differentiation and activation of immune sup-
pressor cells, they modulate antigen presentation, and are
able to induce T-cell apoptosis. Although cancer-derived
exosomes mainly suppress the immune system and facili-
tate tumor progression, they are also important sources of
tumor antigens with potential clinical application in stimu-
lating immune responses. This review summarizes how
exosomes assist cancer to escape immune recognition and
to acquire control over the immune system.
Keywords Exosomes · Extracellular vesicles · Cancer
immunosuppression · Suppressor cells · Immune escape