Innate Lymphoid Cells in Type 2 Immune Responses
Ananda S. Mirchandani • Robert J. Salmond
Abstract In recent years, several distinct innate lymphoid cell populations (ILC) have been characterized in mice and humans. Group 2 ILC function as a rapid responder population in type 2 immune responses. Thus, a wealth of data has implicated an important role for ILC2 in immunity to parasitic infection and in immune pathology in inflammatory and allergic responses. In this review, we describe recent progress in our understanding of the development and ontogeny of ILC2 populations and the mechanisms by which these cells function in a variety of infection and disease settings. Finally, we emphasize recent findings indicating functional interactions between these innate cells and their adaptive CD4? Th2 cell counterparts.
Keywords Innate lymphoid cell Th2 Parasite Allergy
Transcription Factors and Epigenetic Modulation: Its Therapeutic Implication in Chronic Kidney Disease
Kaori Hayashi • Hiroshi Itoh
Abstract Recently emerging evidence has shown that epigenetic mechanisms are involved in initiation and progression of various diseases, including kidney diseases. In the present article, we review the current data regarding the role of epigenetic modulation in chronic kidney disease (CKD) and kidney fibrosis, including DNA methylation and histone modification. Especially we focused on the role of transcription factors in epigenetic modulation and the possibility of therapeutic target of CKD. We have recently reported that transcription factor Kruppel-like factor 4 (also known as gutenriched Kruppel-like factor) is expressed in kidney podocytes (visceral epithelial cells) and modulates podocyte phenotype by gene-selective epigenetic control. Targeting transcription factors for epigenetic modification may be a good candidate for remission and regression of CKD. It is necessary for the therapy of CKD with an epigenetic-based approach to investigate organ-, tissue-, or gene-specific treatment methods for reduction of side effects.
Keywords Chronic kidney disease Epigenetics DNA methylation Histone modification Transcription factors
Expression of the Receptor for Advanced Glycation End Products,
a Target for High Mobility Group Box 1 Protein, and its Role
in Chronic Recalcitrant Rhinosinusitis with Nasal Polyps
Karolina Dzaman • Miroslaw J. Szczepanski •
Marta Molinska-Glura • Antoni Krzeski •
Mariola Zagor
Received: 8 February 2014 / Accepted: 22 September 2014 / Published online: 12 December 2014
Ó The Author(s) 2014. This article is published with open access at Springerlink.com
Abstract A receptor for advanced glycation end products
(RAGE) and its ligand high mobility group box 1
(HMGB1) protein has been linked to several chronic dis-
eases, and acts as a trigger for inflammation signaling.
Here, we study RAGE and HMGB1 expression in chronic,
recalcitrant rhinosinusitis with nasal polyps (CRSwNP) to
determine its potential clinical significance, i.e., disease
recurrence and severity. RAGE and HMGB1 expression in
CRSwNP was evaluated by immunohistochemistry in
epithelial cells of fresh sinonasal mucosa samples obtained
from the patients diagnosed with recalcitrant CRSwNP
(n = 25) and normal control mucosa (NC) (n = 26).
RAGE and HMGB1 expression levels in tissues were
correlated with disease severity assessed by nasal endos-
copy, CT scan, number of previous sinus surgeries, allergy
status and nasosinusal microbiology. RAGE and HMGB1
were moderately or strongly expressed in CRSwNP tissue.
No or weak RAGE expression was found in NC. HMGB1
was equally strongly expressed in NC. We observed a
strong correlation between RAGE and disease severity,
recurrence, undergone operations, asthma and aspirin
exacerbated respiratory disease (AERD). Elevated RAGE
expression is associated with increased disease severity, as
well as allergy and AERD in patients with recalcitrant
CRSwNP. It is possible that the explanation for recurrent
CRSwNP pathogenesis might be related to RAGE
overexpression with subsequent sinus mucosa hyperpro-
liferation, necessitating several operations.
Keywords Nasal polyposis Innate immunity
Chronic inflammation RAGE HMGB1
Transmission-Blocking Vaccines: Focus on Anti-Vector Vaccines
against Tick-Borne Diseases
Girish Neelakanta • Hameeda Sultana
Received: 26 August 2014 / Accepted: 15 October 2014 / Published online: 12 December 2014
Ó The Author(s) 2014. This article is published with open access at Springerlink.com
Abstract Tick-borne diseases are a potential threat that
account for significant morbidity and mortality in human
population worldwide. Vaccines are not available to treat
several of the tick-borne diseases. With the emergence and
resurgence of several tick-borne diseases, emphasis on the
development of transmission-blocking vaccines remains
increasing. In this review, we provide a snap shot on some
of the potential candidates for the development of anti-
vector vaccines (a form of transmission-blocking vaccines)
against wide range of hard and soft ticks that include
Ixodes, Haemaphysalis, Dermacentor, Amblyomma, Rhi-
picephalus and Ornithodoros species.
Keywords Transmission-blocking vaccine
Anti-vector vaccine Ixodes Dermacentor Amblyomma
Haemaphysalis Rhipicephalus Ornithodoros species
Stem Cell-Based Approaches to Improve Nerve Regeneration:
Potential Implications for Reconstructive Transplantation?
Saami Khalifian • Karim A. Sarhane • Markus Tammia •
Zuhaib Ibrahim • Hai-Quan Mao • Damon S. Cooney •
Jaimie T. Shores • W. P. Andrew Lee • Gerald Brandacher
Received: 28 August 2013 / Accepted: 7 October 2014 / Published online: 27 November 2014
Ó L. Hirszfeld Institute of Immunology and Experimental Therapy, Wroclaw, Poland 2014
Abstract Reconstructive transplantation has become a
viable option to restore form and function after devastating
tissue loss. Functional recovery is a key determinant of
overall success and critically depends on the quality and
pace of nerve regeneration. Several molecular and cell-
based therapies have been postulated and tested in pre-
clinical animal models to enhance nerve regeneration.
Schwann cells remain the mainstay of research focus pro-
viding neurotrophic support and signaling cues for
regenerating axons. Alternative cell sources such as mes-
enchymal stem cells and adipose-derived stromal cells
have also been tested in pre-clinical animal models and in
clinical trials due to their relative ease of harvest, rapid
expansion in vitro, minimal immunogenicity, and capacity
to integrate and survive within host tissues, thereby over-
coming many of the challenges faced by culturing of
human Schwann cells and nerve allografting. Induced
pluripotent stem cell-derived Schwann cells are of partic-
ular interest since they can provide abundant, patient-
specific autologous Schwann cells. The majority of
experimental evidence on cell-based therapies, however,
has been generated using stem cell-seeded nerve guides
that were developed to enhance nerve regeneration across
‘‘gaps’’ in neural repair. Although primary end-to-end
repair is the preferred method of neurorrhaphy in recon-
structive transplantation, mechanistic studies elucidating
the principles of cell-based therapies from nerve guidance
conduits will form the foundation of further research
employing stem cells in end-to-end repair of donor and
recipient nerves. This review presents key components of
nerve regeneration in reconstructive transplantation and
highlights the pre-clinical studies that utilize stem cells to
enhance nerve regeneration.
Keywords Reconstructive transplantation
Vascularized composite allotransplantation
Nerve regeneration Functional recovery
Stem cells Schwann cells
Human Antibody Production in Transgenic Animals
Marianne Bru¨ ggemann • Michael J. Osborn •
Biao Ma • Jasvinder Hayre • Suzanne Avis •
Brian Lundstrom • Roland Buelow
Received: 18 November 2014 / Accepted: 19 November 2014 / Published online: 3 December 2014
Ó The Author(s) 2014. This article is published with open access at Springerlink.com
Abstract Fully human antibodies from transgenic ani-
mals account for an increasing number of new therapeutics.
After immunization, diverse human monoclonal antibodies
of high affinity can be obtained from transgenic rodents,
while large animals, such as transchromosomic cattle, have
produced respectable amounts of specific human immu-
noglobulin (Ig) in serum. Several strategies to derive
animals expressing human antibody repertoires have been
successful. In rodents, gene loci on bacterial artificial
chromosomes or yeast artificial chromosomes were inte-
grated by oocyte microinjection or transfection of
embryonic stem (ES) cells, while ruminants were derived
from manipulated fibroblasts with integrated human chro-
mosome fragments or human artificial chromosomes. In all
strains, the endogenous Ig loci have been silenced by gene
targeting, either in ES or fibroblast cells, or by zinc finger
technology via DNA microinjection; this was essential for
optimal production. However, comparisons showed that
fully human antibodies were not as efficiently produced as
wild-type Ig. This suboptimal performance, with respect to
immune response and antibody yield, was attributed to
imperfect interaction of the human constant region with
endogenous signaling components such as the Iga/b in
mouse, rat or cattle. Significant improvements were
obtained when the human V-region genes were linked to
the endogenous CH-region, either on large constructs or,
separately, by site-specific integration, which could also
silence the endogenous Ig locus by gene replacement or
inversion. In animals with knocked-out endogenous Ig loci
and integrated large IgH loci, containing many human Vs,
all D and all J segments linked to endogenous C genes,
highly diverse human antibody production similar to nor-
mal animals was obtained.
Keywords Transgenic animals Antibody repertoires
Human epitopes Class-switch recombination
B cell development Locus silencing
Erratum to: Neutrophil Myeloperoxidase: Soldier and Statesman
Zofia Prokopowicz • Janusz Marcinkiewicz •
David R. Katz • Benjamin M. Chain
Published online: 26 November 2014
Ó L. Hirszfeld Institute of Immunology and Experimental Therapy, Wroclaw, Poland 2014
Erratum to: Arch. Immunol. Ther. Exp. (2012) 60:43–54
DOI 10.1007/s00005-011-0156-8
Unfortunately, the Acknowledgments section has been
published as blinded version. The complete version is
given below:
Authors thank Dr. Rafał Biedron´ for assistance in the
preparation of this manuscript for publication. Zofia Pro-
kopowicz was supported by a MRC studentship. Benjamin
Chain is supported by a grant from Ovarian Cancer Action.
Janusz Marcinkiewicz is supported by a grant from the
Jagiellonian University Medical College (Grant No.
K/ZDS/001008) and Grant NCN no. N N401 042139.
Incidence of Adenoviral DNAemia in Polish Adults Undergoing
Allogeneic Haematopoietic Stem Cell Transplantation
Sylwia Rynans • Tomasz Dziecia˛tkowski • Maciej Przybylski •
Grzegorz W. Basak • Patrycja Rusicka • Agnieszka Tomaszewska •
Kazimierz Hałaburda • Wiesław W. Je˛drzejczak • Gra_zyna Młynarczyk
Received: 28 November 2013 / Accepted: 12 August 2014 / Published online: 7 November 2014
Ó L. Hirszfeld Institute of Immunology and Experimental Therapy, Wroclaw, Poland 2014
Abstract Human adenoviruses (HAdV) are important
viral pathogens recognized increasingly in immunocom-
promised hosts, especially in allogeneic haematopoietic
stem cell transplant recipients (alloHSCT). The clinical
spectrum of HAdV disease ranges from asymptomatic vi-
raemia and mild self-limiting disease to lower respiratory
tract infection, multi-organ involvement and even death.
Early detection and quantification of HAdV in peripheral
blood using real-time PCR (qPCR) assay has been suggested
as a useful monitoring tool, but is seldom used for regular
surveillance of HAdV in haematology centers. A group of
112 alloHSCT recipients from two hospitals in Warsaw
(Poland) was examined in the early post-transplant period
using a quantitative qPCR assay. A total of 1,245 serum
samples were evaluated for presence of HAdV DNA in
patients where 66 (59 %) patients received grafts from
unrelated donors whereas the other 46 (41 %) from sibling
donors. HAdV sequences were detected in 64 (57 %) of the
112 patients. In 22 of all patients (20 %) HAdV DNA was
detected only in a single positive sample, while 42 (37 %)
had positive results in two or more subsequent sera. In total,
DNAemia was present in 202 sera samples (16 %) with
median time to observation of 47 days. Graft-versus-host
disease (GvHD) was observed in 18 (28 %) adenovirus-
infected transplant recipients and a significant correlation
between HAdV infections and GvHD clinical presentation
was found (p = 0.018). There is a high prevalence of HAdV
infections in HSCT recipients in Poland during early post-
transplant period. In consequence, we could only speculate if
HAdV DNAemia could be also related to GvHD symptoms,
enforcing the important pathogenic role of these viral
infections in clinical complications post-alloHSCT.
Keywords HAdV
Haematopoietic stem cell transplantation
Post-transplant infectious complications Real-time PCR
IL–17A, IL–17F and IL–23R Gene Polymorphisms in Polish
Patients with Rheumatoid Arthritis
Katarzyna Bogunia-Kubik • Jerzy S´wierkot • Anna Malak •
Barbara Wysoczan´ ska • Beata Nowak • Katarzyna Białowa˛s •
Katarzyna Ge˛bura • Lucyna Korman • Piotr Wiland
Received: 24 February 2014 / Accepted: 27 August 2014 / Published online: 12 November 2014
Ó The Author(s) 2014. This article is published with open access at Springerlink.com
Abstract Among the complex network of inflammatory
cells involved in the pathogenesis of rheumatoid arthritis
(RA), Th17 cells have recently been identified as key cells in
the promotion of autoimmune processes, and joint destruc-
tion. The IL-23/Th17 signalling pathway, consisting of IL-
23/IL-23R, IL-17A and IL-17F encoding genes, represents a
candidate way for RA development with possible involve-
ment in disease susceptibility and effect on disease
progression. The present study aimed to determine the
association between the polymorphic variants of the IL–17A
(rs2275913), IL–17F (rs763780) and IL–23R (rs11209026)
genes and RA susceptibility, progression and response to
therapy with TNF-a inhibitors. Eighty-nine patients and 125
healthy individuals were investigated. The IL–17A poly-
morphism was found to affect RA progression and response
to anti-TNF treatment. Female patients carrying the IL–17A
wild-type genotype more frequently presented with stage 4
(8/24 vs. 6/47; p = 0.058) and were characterized by more
active disease (the highest DAS28 score [5.1) after
3 months of therapy with the TNF inhibitors (12/23 vs.
15/45; p = 0.040). The IL–17F polymorphism appeared to
be associated with susceptibility to the disease. The presence
of the IL–17F minor variant (OR 3.97; p \ 0.001) and its
homozygosity (OR 29.62; p \ 0.001) was more frequent
among patients than healthy individuals. These results sug-
gest that the polymorphisms within the IL–17A and IL–17F
genes play a significant role in RA.
Keywords Th17 IL-17A IL-17F IL-23R
Gene polymorphism Rheumatoid arthritis
Disease progression Therapy with TNF-alpha inhibitors
Peptide Receptor Radionuclide Therapy of Differentiated Thyroid
Cancer: Efficacy and Toxicity
Rafał Czepczyn´ ski • Magdalena Matysiak-Grzes´ • Maria Gryczyn´ ska •
Maciej Ba˛czyk • Anna Wyszomirska • Marek Stajgis • Marek Ruchała
Received: 17 January 2014 / Accepted: 1 August 2014 / Published online: 18 November 2014
Ó The Author(s) 2014. This article is published with open access at Springerlink.com
Abstract In rare cases of differentiated thyroid carci-
noma (DTC), radioiodine treatment is no longer effective
due to cell dedifferentiation. Targeting somatostatin
receptors in DTC cells by radiolabelled somatostatin ana-
logues could provide an alternative therapy option. The
aim of this study was to evaluate safety and efficacy of
peptide receptor radionuclide therapy (PRRT) in patients
with advanced, non-iodine avid DTC. Eleven patients aged
47–81 years (median: 65 years) with a history of several
courses of radioiodine therapy, increasing thyroglobulin
(Tg) and negative whole body scan, were qualified to the
study. After confirming receptor expression by somato-
statin receptor scintigraphy, PRRT with yttrium-90 labelled
analogue was initiated. Fractionated treatment protocol was
used with four doses of 90Y-DOTA-TOC in 12-week
intervals. Activity of each dose was 3.7 GBq (100 mCi).
Of 11 patients, 5 died before receiving the fourth course of
PRRT. In the remaining six patients, morphological
response, evaluated 3 months after the last course using
RECIST criteria showed partial remission (PR) in one
patient, stable disease (SD) in two patients and progressive
disease (PD) in three patients. Biochemical response based
on Tg measurements before and after PRRT showed PR in
one patient, SD in four patients and PD in one patient.
Median survival was 21 months from the first course of
PRRT. Only minor and transient hematological toxicity
was observed in some patients. We conclude that PRRT is
generally well-tolerated and may be a valuable option for
some patients with radioiodine-refractory DTC.
Keywords Thyroid cancer Peptide receptor
radionuclide therapy Efficacy Toxicity
Immune Modulation in Xenotransplantation
Magdalena Boksa • Joanna Zeyland •
Ryszard Słomski • Daniel Lipin´ ski
Received: 7 February 2014 / Accepted: 22 July 2014 / Published online: 30 October 2014
Ó The Author(s) 2014. This article is published with open access at Springerlink.com
Abstract The use of animals as donors of tissues and
organs for xenotransplantations may help in meeting the
increasing demand for organs for human transplantations.
Clinical studies indicate that the domestic pig best satisfies
the criteria of organ suitability for xenotransplantation.
However, the considerable phylogenetic distance between
humans and the pig causes tremendous immunological
problems after transplantation, thus genetic modifications
need to be introduced to the porcine genome, with the aim
of reducing xenotransplant immunogenicity. Advances in
genetic engineering have facilitated the incorporation of
human genes regulating the complement into the porcine
genome, knockout of the gene encoding the formation of
the Gal antigen (a1,3-galactosyltransferase) or modifica-
tion of surface proteins in donor cells. The next step is two-
fold. Firstly, to inhibit processes of cell-mediated xenograft
rejection, involving natural killer cells and macrophages.
Secondly, to inhibit rejection caused by the incompatibility
of proteins participating in the regulation of the coagulation
system, which leads to a disruption of the equilibrium in
pro- and anti-coagulant activity. Only a simultaneous
incorporation of several gene constructs will make it pos-
sible to produce multitransgenic animals whose organs,
when transplanted to human recipients, would be resistant
to hyperacute and delayed xenograft rejection.
Keywords Xenotransplantation Transgenic pig
Hyperacute rejection Cytotoxicity Coagulation
Natural Foci Diseases as a Stable Biological Threat
Nataliya Vynograd
Received: 28 February 2014 / Accepted: 10 October 2014 / Published online: 19 October 2014
Ó L. Hirszfeld Institute of Immunology and Experimental Therapy, Wroclaw, Poland 2014
Abstract The key aspects of the natural foci of especially
dangerous diseases as a type of biological threats are pre-
sented. Approaches to epidemiological surveillance and
control to the spread of the agents of especially dangerous
diseases on endemic areas are described for zoonosis that
has a medical value. The knowledge of specific design of
tools for the implementation of epidemiological surveil-
lance, monitoring and evaluation of natural foci diseases in
developing countries is low; accordingly, little is known on
the ecology and transmission dynamics for the agents of
especially dangerous diseases. Important is to know the
effectiveness of serological monitoring of the indigenous
population to determine the activity of natural foci of
hemorrhagic fever with renal syndrome, tick-borne
encephalitis, tularemia, Q-fever, Lyme disease and West
Nile disease. The main species of reservoirs and vectors for
these agents have been determined in different regions of
Ukraine. New tick-borne agents that were unknown for
certain regions have been detected. These data indicate the
spreading of different pathogens in combination with nat-
ural foci.
Keywords Natural foci Especially dangerous diseases
Differential Inflammatory MicroRNA and Cytokine Expression
in Pulmonary Sarcoidosis
Agnieszka Jazwa • Lukasz Kasper • Maciej Bak •
Mateusz Sobczak • Krzysztof Szade • Alicja Jozkowicz •
Krzysztof Sladek • Jozef Dulak
Received: 12 March 2014 / Accepted: 5 August 2014 / Published online: 1 November 2014
Ó The Author(s) 2014. This article is published with open access at Springerlink.com
Abstract Sarcoidosis is a granulomatous disease of
unknown etiology. The disease has an important inflam-
matory and immune component; however, its
immunopathogenesis is not completely understood.
Recently, the role of microRNAs (miRNAs), the small non-
coding RNAs, has attracted attention as both being
involved in pathogenesis and serving as disease markers.
Accordingly, changes in the expression of some miRNAs
have been also associated with different autoimmune
pathologies. However, not much is known about the role of
miRNAs in sarcoidosis. Therefore, the aim of this study
was to compare the level of expression of selected miRNAs
in healthy individuals and patients with sarcoidosis. We
detected significantly increased level of miR-34a in
peripheral blood mononuclear cells isolated from sarcoid-
osis patients. Moreover, significantly up-regulated levels of
interferon (IFN)-c, IFN-c inducible protein (IP-10) and
vascular endothelial growth factor were detected in sera of
patients when compared to healthy subjects. Our results
add to a known inflammatory component in sarcoidosis.
Changes in the levels of miR-34a may suggest its
involvement in the pathology of this disease.
Keywords Sarcoidosis miRNA miR-34a IFN-c
Vascular endothelial growth factor Inflammation
Colonization of Bordetella pertussis Clinical Isolates that Differ
by Pulsed Field Gel Electrophoresis Types in the Lungs of Naı¨ve
Mice or Mice Immunized with the Whole-Cell Pertussis Vaccine
Used in Poland
Maciej Polak • Monika Zawadka • Ewa Mosiej • Daniel Rabczenko •
Ewa Augustynowicz • Nicole Guiso • Anna Lutyn´ ska
Received: 13 March 2014 / Accepted: 18 June 2014 / Published online: 9 October 2014
Ó L. Hirszfeld Institute of Immunology and Experimental Therapy, Wroclaw, Poland 2014
Abstract The goal of our study was to compare the
elimination of Bordetella pertussis clinical isolates that
differ according to pulsed field gel electrophoresis (PFGE),
serotypes and genes encoding virulence factors from the
lungs of naı¨ve mice or mice immunized with commercial
diphtheria-tetanus-whole-cell pertussis vaccine used in
Poland. When a mixture of four isolates, given in equal
proportions and harboring different PFGE profiles, sero-
types, and alleles encoding virulence factors, was used to
infect non-immunized mice, a single isolate, characterized
by PFGE type IVc, Fim2 phenotype and ptxA1–prn2–
tcfA2–fim2-1–ptxP1–ptxC1–fim3-1 alleles, was found to be
significantly predominant compared to the others. This
PFGE profile is commonly found in B. pertussis isolates
circulating in some European countries since the late
1990s, confirming its high fitness. The Polish commercial
whole-cell pertussis vaccine induced an immunity effective
at eliminating the B. pertussis isolates from the lungs.
However, the elimination of the isolate harboring PFGE
type C profile, Fim2,3 phenotype and ptxA1–prn1–tcfA2–
fim2-1–ptxP1–ptxC1–fim3-1 alleles was delayed as com-
pared to the others, suggesting phenotypic differences with
the other isolates and vaccine strains. Nevertheless, the
same isolate, when challenged into mice in the defined
mixture of strains, lost the competition with the others, as
measured by lung colonization efficiency. This PFGE
profile represents 15 % of the isolates circulating in Poland
between 2001 and 2012.
Keywords Bordetella pertussis Whole-cell
pertussis vaccine PFGE Pertussis toxin S1 subunit
Pertactin Mouse model of infection
Chemokines and Skin Diseases
Makoto Sugaya
Received: 29 May 2014 / Accepted: 26 August 2014 / Published online: 3 September 2014
Ó L. Hirszfeld Institute of Immunology and Experimental Therapy, Wroclaw, Poland 2014
Abstract Chemokines are small molecules that induce
chemotaxis and activation of certain subsets of leukocytes.
The expression patterns of chemokines and chemokine
receptors are specific to certain organs and cells. Therefore,
chemokines are important to elucidate the mechanism of
organ-specific human diseases. CCL17 expressed by Lan-
gerhans cells, blood endothelial cells, and fibroblasts plays
a key role in attracting Th2 cells and tumor cells of adult
T-cell leukemia/lymphoma and mycosis fungoides/Se´zary
syndrome into the skin, developing various Th2-type
inflammatory skin diseases as well as cutaneous lym-
phoma. CCL11 and CCL26 expressed by skin-resident
cells, such as fibroblasts, blood endothelial cells, and
keratinocytes, induce infiltration of CCR3-expressing cells
such as Th2 cells and eosinophils. CCL11 may also serve
as an autocrine as well as a paracrine in anaplastic large
cell lymphoma. CX3CL1 expressed on blood endothelial
cells leads to infiltration of CX3CR1? immune cells, such
as mast cells, neutrophils, and macrophages, playing
important roles in wound healing, tumor immunity, and
vasculitis. Biologics targeting chemokines and their
receptors are promising strategies for various skin diseases
that are resistant to the current therapy.
Keywords CCL17 CCR4 CCL11 CCL26
CCR3 CX3CL1
Plasma Kynurenic Acid Concentration in Patients Undergoing
Cardiac Surgery: Effect of Anaesthesia
Edyta Kotlinska-Hasiec • Patrycja Nowicka-Stazka •
Jolanta Parada-Turska • Krzysztof Stazka • Janusz Stazka •
Przemyslaw Zadora • Wojciech Dabrowski
Received: 24 March 2014 / Accepted: 20 May 2014 / Published online: 10 September 2014
Ó The Author(s) 2014. This article is published with open access at Springerlink.com
Abstract Increases in plasma kynurenic acid (KYNA)
concentration relate to the severity of inflammation. The
aim of this study was to analyse changes in plasma KYNA
concentration and neutrophil/lymphocyte ratio (NLR) in
cardiac surgery patients. Additionally, the effect of anaes-
thesia was analysed. Adult cardiac surgery patients under
intravenous general anaesthesia were studied. Additionally,
some patients received sevoflurane (SEV) prior to cardio-
pulmonary bypass. Plasma KYNA concentration and NLR
were measured before anaesthesia, just after surgery and on
postoperative days 1, 2 and 3. Patients were assigned to
two groups: patients who did not receive SEV (NonSEV
group) and patients who received SEV (SEV group). Forty-
three patients were studied. Twenty-four of them received
SEV. KYNA increased immediately after surgery and
remained elevated through postoperative day 3 in the
NonSEV group, whereas it was similar to the preoperative
concentration in the SEV group. NLR increased immedi-
ately after surgery in both groups, and higher values were
noted in the NonSEV group than in the SEV group at
postoperative days 2 and 3. Plasma KYNA concentration
correlated with NLR in the NonSEV group. Cardiac sur-
gery caused an increase in NLR. Plasma KYNA increased
in the NonSEV group and correlated with NLR. Admin-
istration of SEV inhibited the increase in KYNA, most
likely due to its anti-inflammatory properties.
Keywords Kynurenic acid
Neutrophil/lymphocyte ratio Sevoflurane
Cardiac surgery General anaesthesia
Opioids, Neutral Endopeptidase, its Inhibitors and Cancer: Is
There a Relationship among them?
Magdalena Mizerska-Dudka • Martyna Kandefer-Szerszen´
Received: 11 March 2014 / Accepted: 18 June 2014 / Published online: 6 September 2014
Ó The Author(s) 2014. This article is published with open access at Springerlink.com
Abstract The role of endogenous animal opioids in the
biology of cancer is widely recognized but poorly under-
stood. This is, among others, because of the short half-life
of these peptides, which are quickly inactivated by endo-
peptidases, e.g., neutral endopeptidase (NEP, CD10). It has
been established that NEP is engaged in the modulation of
the tumor microenvironment, among others that of colon
cancer, by exerting influence on cell growth factors, the
extracellular matrix and other biologically active sub-
stances. Although there are some discrepancies among the
findings on the role of both opioids and NEP in cancer
development, authors agree that their role seems to depend
on the origin, stage and grade of tumor, and even on the
method of examination. Moreover, recently, natural
inhibitors of NEP, such as sialorphin, opiorphin and spin-
orphin have been detected. Their analgesic activity has
been established. It is interesting to ask whether there is a
relationship among opioid peptides, tumor-associated NEP
and its inhibitors.
Keywords Opioid peptides OGF Colon cancer
Neutral endopeptidase CD10
The Role of Endothelin-1 and Endothelin Receptor Antagonists
in Inflammatory Response and Sepsis
Agata Kowalczyk • Paulina Kleniewska •
Michal Kolodziejczyk • Beata Skibska •
Anna Goraca
Received: 13 November 2013 / Accepted: 18 July 2014 / Published online: 7 October 2014
Ó The Author(s) 2014. This article is published with open access at Springerlink.com
Abstract Endothelin-1 (ET-1) is a potent endogenous
vasoconstrictor, mainly secreted by endothelial cells. It acts
through two types of receptors: ETA and ETB. Apart from
a vasoconstrictive action, ET-1 causes fibrosis of the vas-
cular cells and stimulates production of reactive oxygen
species. It is claimed that ET-1 induces proinflammatory
mechanisms, increasing superoxide anion production and
cytokine secretion. A recent study has shown that ET-1 is
involved in the activation of transcription factors such as
NF-jB and expression of proinflammatory cytokines
including TNF-a, IL-1, and IL-6. It has been also indicated
that during endotoxaemia, the plasma level of ET-1 is
increased in various animal species. Some authors indicate
a clear correlation between endothelin plasma level and
morbidity/mortality rate in septic patients. These patho-
logical effects of ET-1 may be abrogated at least partly by
endothelin receptor blockade. ET-1 receptor antagonists
may be useful for prevention of various vascular diseases.
This review summarises the current knowledge regarding
endothelin receptor antagonists and the role of ET-1 in
sepsis and inflammation.
Keywords Endothelins Sepsis Inflammation
Reactive oxygen species Endothelin receptor antagonists
Cyclosporine A Regulates Pro-Inflammatory Cytokine Production
in Ulcerative Colitis
Stefanie Steiner • Carolin Daniel • Anika Fischer • Imke Atreya •
Simon Hirschmann • Maximilian Waldner • Helmut Neumann •
Markus Neurath • Raja Atreya • Benno Weigmann
Received: 22 November 2013 / Accepted: 23 May 2014 / Published online: 26 August 2014
Ó L. Hirszfeld Institute of Immunology and Experimental Therapy, Wroclaw, Poland 2014
Abstract Crohn’s disease (CD) and ulcerative colitis
(UC) are the two major forms of inflammatory bowel
diseases (IBD), which are defined as relapsing inflamma-
tions of the gastrointestinal tract. Cyclosporine A (CsA) is
a potential rescue treatment to avoid colectomy in severe
steroid-refractory UC patients. The molecular mechanism
of action of CsA in UC is nevertheless still not well
understood. The aim of this study was to investigate the
effect of CsA on a possible modulation of cytokine pro-
duction by peripheral blood mononuclear cells (PBMCs) of
controls and patients with UC or CD. Upon CsA treatment,
analyses of cytokine levels revealed a significant reduction
of IL-13 expression in PBMCs from patients with UC,
whereas other cytokine expression levels remained unaf-
fected. To address the question whether CsA treatment
impinges on the induction of cell death, apoptosis assays
were performed using CD4? T cells from peripheral blood
of patients suffering from either UC or CD. It became clear
that CsA treatment resulted in a specific induction of
apoptosis in samples from controls and patients with UC
but not with CD. Apoptosis induction was not mediated via
the mitochondrial apoptosis pathway. The present data
support the concept that CsA treatment modulates pro-
inflammatory cytokine production and T cell survival in
UC via the induction of apoptosis and might therefore help
to explain the clinical efficacy of CsA in patients with UC.
Keywords IBD Apoptosis Cyclosporine A
Ulcerative colitis IL-13
Impact of Imiglucerase Supply Shortage on Clinical
and Laboratory Parameters in Norrbottnian Patients
with Gaucher Disease Type 3
Maciej Machaczka • Cecilia Ka¨mpe Bjo¨rkvall • Joanna Wieremiejczyk •
Martin Paucar Arce • Kristina Myhr-Eriksson • Monika Klimkowska •
Hans Ha¨gglund • Per Svenningsson
Received: 16 December 2013 / Accepted: 7 August 2014 / Published online: 10 September 2014
Ó The Author(s) 2014. This article is published with open access at Springerlink.com
Abstract A viral contamination of the production plant
producing imiglucerase (CerezymeTM) resulted in an un-
predicted worldwide shortage of global supplies during
2009–2010. The aim of the study was to describe the
effects of dose reduction of enzyme replacement therapy
(ERT) in adults with Norrbottnian form of Gaucher disease
type 3 (N-GD3). There were ten adults with N-GD3 treated
with imiglucerase in the county of Norrbotten in June 2009.
Analyzed variables included plasma chitotriosidase activity
and concentration of CCL18/PARC, whole blood hemo-
globin concentration (Hb) and platelet count (PLT), as well
as patients’ body weight, subjective complaints and health
status measured by the EuroQoL-5D questionnaire. The
median duration of ERT shortage lasted for 14 months
(10–20 months). The median percentage reduction of im-
iglucerase dose was 36 % (26–59 %). Hb decreased in four
patients, PLT decreased in three patients, chitotriosidase
increased in three patients (max. ?22 % of baseline), and
CCL18/PARC increased in six patients (?14 % to
?57 %). The body weight was moderately decreased in
one patient. No new bone events were noted. Self-assess-
ment of individual patient’s health status was stable in all
but one patient. Our results suggest that moderate reduction
of ERT dosage lasting for relatively short period of time
can lead to worsening in biomarkers of adults with N-GD3.
However, this worsening is infrequently translated to
clinical worsening of patients. It is possible that CCL18/
PARC has a higher sensitivity than chitotriosidase in
monitoring of ERT dosing in GD3.
Keywords CerezymeTM Enzyme replacement therapy
Gaucher disease Imiglucerase Norrbottnian type
Shortage Supply
Restoring TRAIL Mediated Signaling in Ovarian Cancer Cells
Ammad Ahmad Farooqi • Ilhan Yaylim • Nazlı Ezgi Ozkan •
Farrukh Zaman • Talha Abdul Halim • Hsueh-Wei Chang
Received: 1 August 2013 / Accepted: 26 June 2014 / Published online: 17 July 2014
Ó L. Hirszfeld Institute of Immunology and Experimental Therapy, Wroclaw, Poland 2014
Abstract Ovarian cancer has emerged as a multifaceted
and genomically complex disease. Genetic/epigenetic
mutations, suppression of tumor suppressors, overexpres-
sion of oncogenes, rewiring of intracellular signaling
cascades and loss of apoptosis are some of the deeply
studied mechanisms. In vitro and in vivo studies have
highlighted different molecular mechanisms that regulate
tumor necrosis factor-related apoptosis-inducing ligand
(TRAIL) mediated apoptosis in ovarian cancer. In this
review, we bring to limelight, expansion in understanding
systematical characterization of ovarian cancer cells has
led to the rapid development of new drugs and treatments
to target negative regulators of TRAIL mediated signaling
pathway. Wide ranging synthetic and natural agents have
been shown to stimulate mRNA and protein expression of
death receptors. This review is compartmentalized into
programmed cell death protein 4, platelet-derived growth
factor signaling and miRNA control of TRAIL mediated
signaling to ovarian cancer. Mapatumumab and PRO95780
have been tested for efficacy against ovarian cancer. Use of
high-throughput screening assays will aid in dissecting the
heterogeneity of this disease and increasing a long-term
survival which might be achieved by translating rapidly
accumulating information obtained from molecular and
cellular studies to clinic researches.
Keywords TRAIL Ovarian cancer Apoptosis
Economic Impact Profiling of CBRN Events:
Focusing on Biological Incidents
Simona Cavallini • Fabio Bisogni • Marco Mastroianni
Received: 7 March 2014 / Accepted: 9 July 2014 / Published online: 22 July 2014
Ó L. Hirszfeld Institute of Immunology and Experimental Therapy, Wroclaw, Poland 2014
Abstract Chemical, biological, radiological and nuclear
(CBRN) incidents, both caused accidentally by human
error or natural/technological events and determined
intentionally as criminal/malicious/terroristic acts, have
consequences that could be differently characterized. In the
last years many efforts to analyze the economic impact of
terrorist threat have been carried out, while researches
specifically concerning CBRN events have not been
extensively undertaken. This paper in particular aims at
proposing a methodological approach for studying macro-
level economic impact profiles of biological incidents
caused by weaponized and non-weaponized materials. The
suggested approach investigates the economic conse-
quences of biological incidents according to two main
dimensions: type of large-scale effect and persistence of
effect. Biological incident economic impacts are analyzed
taking into account the persistence of effect during time as
short-term impact (i.e. immediately after the incident),
medium-term impact (i.e. by a month) and long-term
impact (i.e. by years). The costs due to preventive coun-
termeasure against biological threats (e.g. prevention,
protection and preparedness expenses) are not taken into
account. To this purpose, information on the key features
of past biological incidents can be used as case studies to
try to build impact profiles taking into account the
proposed two main dimensions. Consequence management
and effect mitigation of CBRN emergencies and disasters
may benefit from an ex ante definition of the impact pro-
filing related to this kind of incidents. The final goal of this
paper is to define an approach to organize information on
possible biological events according to their impact profile
for supporting more effective and efficient first responders’
prompt actions and policy makers’ strategic decisions after
the event occurrence.
Keywords CBRN events Biological incidents
Impact profiles Persistence Economic impact
Molecular and Chemical Engineering of Bacteriophages
for Potential Medical Applications
Katarzyna Hodyra • Krystyna Da˛browska
Received: 28 February 2014 / Accepted: 20 May 2014 / Published online: 22 July 2014
Ó The Author(s) 2014. This article is published with open access at Springerlink.com
Abstract Recent progress in molecular engineering has
contributed to the great progress of medicine. However,
there are still difficult problems constituting a challenge for
molecular biology and biotechnology, e.g. new generation
of anticancer agents, alternative biosensors or vaccines. As
a biotechnological tool, bacteriophages (phages) offer a
promising alternative to traditional approaches. They can
be applied as anticancer agents, novel platforms in vaccine
design, or as target carriers in drug discovery. Phages also
offer solutions for modern cell imaging, biosensor con-
struction or food pathogen detection. Here we present a
review of bacteriophage research as a dynamically devel-
oping field with promising prospects for further
development of medicine and biotechnology.
Keywords Bacteriophages Phage display
Mutagenesis Covalent immobilization Vaccines
Carriers Biosensors
Food Allergy and the Oral Immunotherapy Approach
Carmen M. Cabrera • Jose´ M. Urra
Received: 1 February 2014 / Accepted: 15 May 2014 / Published online: 16 July 2014
Ó L. Hirszfeld Institute of Immunology and Experimental Therapy, Wroclaw, Poland 2014
Abstract Food allergy represents an increasing health
problem, with children being the most affected population.
The symptoms can appear within minutes or hours of
ingesting the offending food, producing skin manifesta-
tions, respiratory, gastrointestinal and anaphylactic
reactions in the severe forms. Food allergy is established by
the loss of tolerance to food proteins, and is characterized
by an altered balance of regulatory T (Treg) cells and the
shift to Th2 type cytokines in the intestinal lamina propria.
We have described the contribution of different factors in
establishing oral tolerance, such as the antigenic exposition
route, the gut microenvironment, and the timing of the food
introduction. Apart from avoiding the food, immunother-
apy is the only intervention which produces oral
desensitization to food proteins. Among the underlying
immunological mechanisms of oral immunotherapy (OIT)
are the changes in humoral immunity (a decrease of
allergen-specific IgE and an increase of allergen-specific
IgG4) and cellular changes such as the increased number of
FoxP3? Treg cells. At present, the experiences of OIT with
various foods are offering promising results. OIT appears
to be safe producing low adverse reactions, and effective in
inducing desensitization in most subjects with food allergy.
Keywords Allergy Food Immunologic mechanisms
Oral tolerance Oral immunotherapy
Novel Transgenic Rice-Based Vaccines
Tatsuhiko Azegami • Hiroshi Itoh • Hiroshi Kiyono •
Yoshikazu Yuki
Received: 28 January 2014 / Accepted: 26 May 2014 / Published online: 16 July 2014
Ó L. Hirszfeld Institute of Immunology and Experimental Therapy, Wroclaw, Poland 2014
Abstract Oral vaccination can induce both systemic and
mucosal antigen-specific immune responses. To control
rampant mucosal infectious diseases, the development of new
effective oral vaccines is needed. Plant-based vaccines are
new candidates for oral vaccines, and have some advantages
over the traditional vaccines in cost, safety, and scalability.
Rice seeds are attractive for vaccine production because of
their stability and resistance to digestion in the stomach. The
efficacy of some rice-based vaccines for infectious, autoim-
mune, and other diseases has been already demonstrated in
animal models. We reported the efficacy in mice, safety, and
stability of a rice-based cholera toxin B subunit vaccine called
MucoRice-CTB. To advance MucoRice-CTB for use in
humans, we also examined its efficacy and safety in primates.
The potential of transgenic rice production as a new mucosal
vaccine delivery system is reviewed from the perspective of
future development of effective oral vaccines.
Keywords Rice-based vaccine MucoRice-CTB
Mucosal immunity Oral vaccine
Biological Agents Database in the Armed Forces
Marcin Niemcewicz • Janusz Kocik •
Anna Bielecka • Michał Wiercin´ ski
Received: 26 February 2014 / Accepted: 18 June 2014 / Published online: 18 July 2014
Ó L. Hirszfeld Institute of Immunology and Experimental Therapy, Wroclaw, Poland 2014
Abstract Rapid detection and identification of the bio-
logical agent during both, natural or deliberate outbreak is
crucial for implementation of appropriate control measures
and procedures in order to mitigate the spread of disease.
Determination of pathogen etiology may not only support
epidemiological investigation and safety of human beings,
but also enhance forensic efforts in pathogen tracing, col-
lection of evidences and correct inference. The article
presents objectives of the Biological Agents Database,
which was developed for the purpose of the Ministry of
National Defense of the Republic of Poland under the
European Defence Agency frame. The Biological Agents
Database is an electronic catalogue of genetic markers of
highly dangerous pathogens and biological agents of weapon
of mass destruction concern, which provides full identifica-
tion of biological threats emerging in Poland and in locations
of activity of Polish troops. The Biological Agents Database
is a supportive tool used for tracing biological agents’ origin
as well as rapid identification of agent causing the disease of
unknown etiology. It also provides support in diagnosis,
analysis, response and exchange of information between
institutions that use information contained in it. Therefore, it
can be used not only for military purposes, but also in a
civilian environment.
Keywords Biological agents Government Military
European defence agency
Caspases as the Key Effectors of Inflammatory Responses Against
Bacterial Infection
Ryosuke Uchiyama • Hiroko Tsutsui
Received: 17 February 2014 / Accepted: 22 May 2014 / Published online: 18 July 2014
Ó L. Hirszfeld Institute of Immunology and Experimental Therapy, Wroclaw, Poland 2014
Abstract Caspase cysteine proteases are factors widely
recognized for their role in the induction of apoptotic cell
death. Caspases induce apoptosis during the inflammatory
response to pathogen infection; in addition, caspases such
as caspase-1 and caspase-11 are known to be involved in
the production of inflammatory cytokines in response to
bacterial infections. Caspase-1 is activated in the inflam-
masome, an intracellular protein complex that is formed by
the recognition of intracellular ligands or cellular stresses
by sensor molecules such as NOD-like receptors. Under
certain conditions, caspase-11 is required for the activation
of the caspase-1 inflammasome, referred to as the non-
canonical inflammasome. In addition to these caspases,
accumulating evidence indicates that caspase-8 also con-
tributes to the production of inflammatory cytokines. In
contrast to caspase-1, caspase-8 is activated by receptors
located on the plasma membrane including dectin-1, TLR-
3/4, and Fas. Recently, Fas-mediated caspase-8 activation
and inflammatory cytokine production have been shown to
play a significant role in the regulation of bacterial infec-
tions. This review highlights the functional roles and
activation mechanisms of caspase-1/-11 in innate immune
responses against bacterial infection. In addition, we dis-
cuss the novel aspects of caspase-8 function in comparison
with caspase-1/-11 during innate inflammatory responses.
Keywords Caspase Inflammasome IL-1b/IL-18
Fas
Gene Polymorphisms of Novel Immunotolerant Molecule BTLA:
Distribution of Alleles, Genotypes and Haplotypes in Polish
Caucasian Population
Anna Partyka • Dariusz Woszczyk • Tomasz Strzała • Anna Szczepan´ ska •
Anna Tomkiewicz • Irena Frydecka • Lidia Karabon
Received: 30 January 2014 / Accepted: 12 May 2014 / Published online: 3 September 2014
Ó The Author(s) 2014. This article is published with open access at Springerlink.com
Abstract B and T lymphocyte attenuator (BTLA) is one
of the members of immunoglobulin superfamily which,
like CTLA-4 and PD-1, is involved in down regulation of
immune response. Despite the important role of BTLA in
maintaining immune homeostasis, relatively little studies
were devoted to the relationship of polymorphisms in the
gene encoding BTLA with susceptibility to autoimmune
disease and cancer. Moreover, all published works were
done in Asian populations. BTLA gene is located on
chromosome 3 in q13.2 and consists of five exons. The aim
of this study was to investigate the alleles, genotypes and
haplotypes frequency of selected BTLA gene polymor-
phisms in Caucasian population originating from Poland.
For this study, the single-nucleotide polymorphisms
(SNPs) were chosen on the basis of literature data. Addi-
tionally, the tag dSNP under linkage equilibrium r2 [ 0.8
and available at the National Center for Biotechnology
Information (NCBI) for Caucasian population of rare
alleles at a frequency greater than 5 % have been chosen
using the NCBI database. The ten BTLA SNPs investigated
were: rs1844089, rs2705535, rs9288952, rs9288953,
rs1982809, rs2633580, rs2705511, rs2705565, rs76844316,
rs16859633. For all SNPs selected on the basis of literature
data the significantly different distributions of genotypes
between Asian and Caucasian populations were observed.
Keywords Gene polymorphism
Co-inhibitory molecule BTLA Caucasian population
Two New Cases of KIR3DP1, KIR2DL4-Negative Genotypes, One
of which is also Lacking KIR3DL2
Wanda Niepiekło-Miniewska • Natalia _Zuk • Joanna Dubis • Maciej Kurpisz •
David Senitzer • Anna Havrylyuk • Ryszard Grendziak • Wojciech Witkiewicz •
Valentyna Chopyak • Piotr Kus´nierczyk
Received: 13 January 2014 / Accepted: 29 April 2014 / Published online: 18 July 2014
Ó The Author(s) 2014. This article is published with open access at Springerlink.com
Abstract The killer immunoglobulin-like receptor (KIR)
genes KIR2DL4, KIR3DL2, and KIR3DP1 are present in
virtually all humans. KIR2DL4 encodes a receptor present
on uterine and decidual natural killer (NK) cells and some
peripheral blood NK cells. Its only known ligand is the
human leukocyte antigen-G molecule expressed on extra-
villous trophoblasts, and on tissues in some diseases.
KIR3DL2 binds HLA-A*03 and HLA-A*11 as well as
HLA-B*27 dimers, and microbial CpG DNA. KIR3DP1 is
a pseudogene. During our immunogenetic studies we found
two individuals, one from Lower Silesia district in Poland,
and another from Western Ukraine, who were reproducibly
negative for KIR2DL4 and KIR3DP1 genes, using three
different PCR systems. Both individuals displayed very
similar genotypes, possessing only KIR3DL3, KIR2DL3,
KIR2DP1, KIR2DS1, and probably a rare variant of
KIR2DL1. The Pole had also KIR3DL2, which the Ukrai-
nian was apparently lacking. The Lower Silesia has been
populated after the Second World War by a remarkable
percentage with displaced people from Western Ukraine,
which might contribute to genetic similarity of the two
individuals described here.
Keywords Killer cell immunoglobulin-like receptor
Framework genes Deletion
Natural or Deliberate Outbreak in Pakistan: How to Prevent
or Detect and Trace its Origin: Biosecurity, Surveillance,
Forensics
Zabta Khan Shinwari • Ali Talha Khalil •
Anwar Nasim
Received: 18 February 2014 / Accepted: 23 May 2014 / Published online: 19 June 2014
Ó L. Hirszfeld Institute of Immunology and Experimental Therapy, Wroclaw, Poland 2014
Abstract Over the last few decades biosecurity and
biosafety have emerged as a prominent public health con-
cern due to some high-profile accidents. Effective
strategies to deal with the outbreak, whether deliberate or
non-deliberate requires a multidisciplinary approach and
coordinated decision-making by various state departments
such as health, forensics, agriculture, environment, intelli-
gence, law and enforcement, etc. In a dynamic global
environment and the overwhelming asymmetric threats
from the non-state actors, it is of utmost importance to
understand the biosecurity issues and initiate a coordinated
global effort to cope with biosecurity and biosafety brea-
ches and develop an as effective response mechanism. An
attractive choice for the terrorists, state enemies and non-
state actors is the use of biological weapons. An unwanted
incident may not only bring chaos to the people, but also
can inflict severe economic damage industrially and locally
as was in the notorious foot-and-mouth disease outbreak.
Because of special geopolitical compulsion, Pakistan is one
of the hot spots where special action needs to be taken. The
current review focuses on the various approaches, tech-
nologies that can be used to alleviate the chances of
biosafety and biosecurity incident and emphasizes the role
of modern technology that can be used in this regard.
Keywords Biosecurity Biosafety Outbreak
Biological weapon
Erratum to: Implant of Polymer Containing Pentacyclic
Triterpenes from Eugenia punicifolia Inhibits Inflammation
and Activates Skeletal Muscle Remodeling
Paulo Emı´lio C. Leite • Katia G. Lima-Arau´ jo •
Guilherme R. Franc¸a • Jussara Lagrota-Candido •
Wilson C. Santos • Thereza Quirico-Santos
Published online: 17 June 2014
Ó L. Hirszfeld Institute of Immunology and Experimental Therapy, Wroclaw, Poland 2014
Erratum to: Arch Immunol Ther Exp
DOI 10.1007/s00005-014-0291-0
Biological Threat Detection in the Air and on the Surface: How
to Define the Risk
El_zbieta Anna Trafny • Rafał Lewandowski •
Małgorzata Ste˛pin´ ska • Miron Kaliszewski
Received: 28 February 2014 / Accepted: 22 May 2014 / Published online: 11 June 2014
Ó L. Hirszfeld Institute of Immunology and Experimental Therapy, Wroclaw, Poland 2014
Abstract The improvements in the existing methods of
rapid detection and biological pathogen surveillance are
still needed. The new spectroscopic methods that rely on
the unique structural features and intrinsic fluorescence of
microorganisms are well fitted for monitoring the spread of
airborne biological agents or their reagentless detection in
the air, and these methods may bring a new quality to
bioaerosols remote detection. This review describes the
problem of the confidence in the environmental testing
results that may affect clearance standard, sampling tech-
niques, and the estimation of risk of human exposure to the
low concentrations of harmful microorganisms during bi-
oterrorist event or naturally occurring outbreaks. Higher
recovery efficiency of dangerous biological agents from the
air and contaminated surfaces would enable more reliable
environmental human risk exposure assessment.
Keywords Bioterrorism Bioaerosols
Stand-off detection Microbial identification
Role of Osteopontin in Systemic Lupus Erythematosus
Beata Kaleta
Received: 29 January 2014 / Accepted: 7 April 2014 / Published online: 11 June 2014
Ó The Author(s) 2014. This article is published with open access at Springerlink.com
Abstract Systemic lupus erythematosus (SLE) is a mul-
tisystemic disease, caused by a variety of factors, which
lead to immunological abnormalities. Osteopontin (OPN)
is a pleiotropic protein, important in bone remodeling and
immune system signaling. OPN, produced by various cells,
including immune cells, plays a key role in regulating
T-helper 1/T-helper 2 balance, stimulating B lymphocytes
to produce antibodies, regulating macrophages, neutrophils
and inducing dendritic cells. OPN expression is influenced
by genetic polymorphisms of its promoter, hormones and
cytokines. Over expression of OPN has been associated
with the pathogenesis of immune-mediated diseases. OPN
has been implicated in the development of murine model of
lupus and in humans with SLE. In this review, I will
present current state of research on the role of OPN and
OPN gene polymorphisms in pathogenesis and clinical
course of SLE. A better understanding of the role of OPN
in SLE will contribute to more precise diagnosis and
treatment of the disease.
Keywords Osteopontin Systemic lupus erythematosus
Gene Polymorphism
Heterogeneity in the Differentiation and Function of CD8+ T Cells
Hans-Willi Mittru¨ cker • Alexander Visekruna •
Magdalena Huber
Received: 19 December 2013 / Accepted: 24 April 2014 / Published online: 31 May 2014
Ó L. Hirszfeld Institute of Immunology and Experimental Therapy, Wroclaw, Poland 2014
Abstract It is well established that CD8? T cells con-
stitute an important branch of adaptive immunity
contributing to clearance of intracellular pathogens and
providing long-term protection. These functions are mostly
fulfilled by the best characterized subpopulation of CD8? T
cells, the cytotoxic T lymphocytes (also called Tc1 cells),
owing to their ability to kill infected cells and to secrete
cytokines such as interferon-c and tumor necrosis factor-a.
However, there is growing evidence for alternative CD8?
T cell fates influencing CD4? T-cell-mediated responses in
the context of allergy, autoimmunity and infections. Thus,
like subpopulations of CD4? T cells, also CD8? T cells
under particular conditions acquire the expression of
interleukin (IL)-4, IL-5, IL-9, IL-13, IL-17 or suppressive
activity and thereby influence immune responses. The
process of CD8? T-cell differentiation is dictated by anti-
gen strength, co-stimulatory molecules and cytokines.
These environmental cues induce transcription factors
further specifying CD8? T-cell decision into Tc1, Tc2,
Tc9, Tc17 or CD8? T regulatory fate. Here, we discuss our
current understanding about functional diversity of effector
CD8? T cells and contribution of transcription factors to
this process.
Keywords CD8? T cells Tc1 Tc2 Tc9 Tc17
CD8? Treg cells
Effect of Diethylcarbamazine Citrate and Setaria equina
Excretory–Secretory Material on Rat Hepatocellular Carcinoma
Mahmoud Abdel-Latif • Thabet Sakran •
Gamal El-Shahawi • Hoda El-Fayoumi •
Al-Mahy El-Mallah
Received: 1 October 2013 / Accepted: 21 March 2014 / Published online: 31 May 2014
Ó L. Hirszfeld Institute of Immunology and Experimental Therapy, Wroclaw, Poland 2014
Abstract Diethylcarbamazine citrate (DEC) has been
known for its efficacy to eradicate bancroftian filariasis in
Egypt and other countries in the world. One of the known
effects was to decrease the level of circulating filarial
antigen in the patient’s serum. The target of this study was
to examine the effect of DEC, excretory–secretory (ES)
material from the filarial parasite Setaria equina or a
combination of both on the status of oxidative stress and
pathogenesis of rat hepatocellular carcinoma (HCC)
induced by diethylnitrosamine and 2-acetylaminofluorene.
This could be tested in vitro using nitroblue tetrazolium
reduction test for measuring the level of superoxide anion
(O2
•–) released from rat peritoneal macrophages. For
in vivo test, a single dose before induction of carcinogen-
esis or continually repeated doses with DEC, ES or
DEC ? ES was used. Exposure of macrophages to ES
could lead to a significant decrease (p \ 0.01) in
O2
•–release, while DEC (200 lM) could modulate such
effect with significant increase (p \ 0.05). Pathogenesis of
liver cancer and treatment were evaluated using histolog-
ical investigation, level of antioxidant and liver function
enzymes. Repeated ES doses could increase the activity of
antioxidant enzymes, especially the catalase enzyme and
show a protective effect on liver architecture. DEC could
modulate the later effects when combined with ES. No
significant effect on the liver function enzymes after
treatment was observed. Nuclear factor jB was found to be
localized only in the cytoplasm after single and repeated
treatments with ES. This study could indicate the effect of
S. equina ES as antioxidant against rat HCC, while DEC
could modulate such effect when combined with it.
Keywords Diethylcarbamazine citrate Setaria equina
Excretory–secretory material Hepatocellular carcinoma
Implant of Polymer Containing Pentacyclic Triterpenes
from Eugenia punicifolia Inhibits Inflammation and Activates
Skeletal Muscle Remodeling
Paulo Emı´lio C. Leite • Katia G. Lima-Arau´ jo •
Guilherme R. Franc¸a • Jussara Lagrota-Candido •
Wilson C. Santos • Thereza Quirico-Santos
Received: 11 June 2013 / Accepted: 24 September 2013 / Published online: 16 May 2014
Ó L. Hirszfeld Institute of Immunology and Experimental Therapy, Wroclaw, Poland 2014
Abstract Sustained chronic inflammation induces acti-
vation of genes involved in cellular proliferation and
apoptosis, thereby causing skeletal muscle degeneration.
To investigate in vitro effects of isolated pentacyclic tri-
terpenes from Eugenia punicifolia (Ep–CM) upon signaling
pathways involved in the regulation of skeletal muscle cell
line proliferation, and in vivo muscular tissue remodeling.
C2C12 cells were seeded on eight-well plates and [3H]-
thymidine incorporation, TUNEL assays, mitochondria
viability, zymography for matrix metalloproteases
(MMPs), Western blot analysis for MAPKinase signaling
pathway, NFjB activation and HMGB1 production sub-
sequently determined under basal conditions and after Ep–
CM treatment. A polymer containing Ep–CM was implan-
ted on the volar surface of gastrocnemius muscles
subjected to acute injury induced by bupivacaine for local
slow and gradual release of bioactive compounds, and mice
killed 4 days after surgery. Ep–CM inhibited proliferation
of C2C12 myoblast cell line in a dose-dependent manner,
confirmed by reduction of [3H]-thymidine uptake without
affecting cell viability or inducing apoptosis. The cytostatic
effect of Ep–CM occurred mainly via inhibition of phos-
phorylated extracellular signal-regulated kinase (pERK)
activation and DNA synthesis, possibly inhibiting the G1
phase of the cell cycle, since Ep–CM increased pAkt and
p27kip1 but reduced Cyclin D1. Ep–CM in vitro treatment
increased MMP-9 and MMP-2 activities of C2C12 myo-
blast cells, but reduced in vivo MMP-9 activity and acute
muscular inflammation. Besides cytostatic and anti-
inflammatory effects, Ep–CM pentacyclic triterpenes also
contributed to degradation of basement membrane com-
ponents by activating mechanisms of skeletal muscle
remodeling in response to local injury.
Keywords Pentacyclic terpenes C2C12 myoblasts
Inflammation Skeletal muscle Muscular injury
Muscle remodeling Bioengineering
State-of-the-Art in Biosafety and Biosecurity in European
Countries
Anna Bielecka • Ali Akbar Mohammadi
Received: 27 January 2014 / Accepted: 8 April 2014 / Published online: 13 May 2014
Ó The Author(s) 2014. This article is published with open access at Springerlink.com
Abstract The terms biosafety and biosecurity are widely
used in different concepts and refer not only to protection
of human beings and their surrounding environment against
hazardous biological agent, but also to global disarmament
of weapons of mass destruction. As a result, the biosafety
and biosecurity issues should be considered interdisci-
plinary based on multilateral agreements against
proliferation of biological weapons, public health and
environmental protection. This publication presents infor-
mation on both, international and national biosafety and
biosecurity legislation. Status of national implementation
of the Biological and Toxin Weapons Convention, penal-
ization issues and measures to account for and secure
production, use, storage of particularly dangerous patho-
gens or activities involving humans, plants and animals
where infection may pose a risk have been analyzed. Safety
and security measures in laboratories have been studied.
Moreover, dual-use technology and measures of secure
transport of biohazard materials have been also taken into
account. In addition, genetic engineering regulations, bio-
security activities in laboratories and code of conducts have
been investigated, as well.
Keywords Biosafety Biosecurity Legislation
BTWC
Designing an Effective Microbial Forensics Program for Law
Enforcement and National Security Purposes
Randall S. Murch
Received: 27 February 2014 / Accepted: 8 April 2014 / Published online: 30 April 2014
Ó L. Hirszfeld Institute of Immunology and Experimental Therapy, Wroclaw, Poland 2014
Abstract Forensic capabilities that provide lead infor-
mation, and investigative, intelligence, prosecution and
policy decision support can be invaluable for responding to
and resolving bioterrorism events. Attributing biological
attacks through scientific and other resources and processes
is an important goal, for which science can be instrumental.
Some even believe that having effective microbial foren-
sics capabilities along with others can even deter
adversaries from using biological weapons. For those
nations that do not have such or wish to integrate or
upgrade capabilities, thoughtful analysis and consideration
of certain design principles will increase the likelihood that
success will be attained.
Keywords Microbial forensics and attribution
Program design principles
Response of Specific Immunoglobulin E to Foods in Children
with Atopic Dermatitis
Susana R. M. Passeti • Fernando L. A. Fonseca •
Neusa F. Wandalsen
Received: 30 July 2013 / Accepted: 6 December 2013 / Published online: 20 April 2014
Ó L. Hirszfeld Institute of Immunology and Experimental Therapy, Wroclaw, Poland 2014
Abstract Food allergy is a common condition that plays
an important role in the pathogenicity and maintenance of
atopic dermatitis (AD), however, must be carefully inves-
tigated before imposing a restrictive diet. The aim of this
study was to evaluate the sensitivity to foods in patients
with AD, correlating it with the severity of the disease and
other possible associated factors. One hundred and eleven
children (6–180 months of age) with AD were evaluated
and later followed up at the Allergy and Clinical Immu-
nology Division, Department of Pediatrics at FMABC. The
serum concentrations of specific IgE to cow’s milk (CM),
egg, soy, wheat, corn, peanut and fish were measured using
an enzymatic fluorescence method (ImmunoCAPTM). In
order to identify the clinical reactivity, the open oral
provocation test was performed when specific IgE was
positive to CM, egg and wheat and in all those who related
symptoms after the intake of such foods regardless of the
presence or absence of sensitization. In total, 40.5 % of the
studied population was sensitized to at least one food
allergen, especially those between 73 and 180 months of
age. There was a higher prevalence of sensitization in
children with more severe AD, and foods like CM, egg and
wheat were the most involved, but with low clinical
reactivity. We observed increased severity of AD in cases
that initiated symptoms earlier and who had shorter dura-
tion of exclusive breastfeeding as well as a linear increase
in sensitization in the most serious cases. Serum-specific
immunoglobulin E was the only factor associated with the
relationship that showed sensitization. The occurrence of
sensitization to foods was frequent, mainly in the age group
of 6–9 years and in patients with severe AD; however, the
validation of the clinical reactivity was negative in most of
the provocations performed, which agrees with the need to
prove the same before the imposition of restrictive diets,
often unnecessary and complex.
Keywords Immunoglobulin E Children
Atopic dermatitis
Ghrelin Gene Products in Acute and Chronic Inflammation
Flavia Prodam • Nicoletta Filigheddu
Received: 29 November 2013 / Accepted: 21 March 2014 / Published online: 12 April 2014
Ó L. Hirszfeld Institute of Immunology and Experimental Therapy, Wroclaw, Poland 2014
Abstract Ghrelin gene products—the peptides ghrelin,
unacylated ghrelin, and obestatin—have several actions on
the immune system, opening new perspectives within
neuroendocrinology, metabolism and inflammation. The
aim of this review is to summarize the available evidence
regarding the less known role of these peptides in the
machinery of inflammation and autoimmunity, outlining
some of their most promising therapeutic applications.
Keywords Acylated ghrelin Unacylated ghrelin
Obestatin Autoimmune disease Inflammatory disease
Circadian Clocks and Inflammation: Reciprocal Regulation
and Shared Mediators
Nicolas Cermakian • Susan Westfall •
Silke Kiessling
Received: 4 October 2013 / Accepted: 22 January 2014 / Published online: 1 April 2014
Ó L. Hirszfeld Institute of Immunology and Experimental Therapy, Wroclaw, Poland 2014
Abstract The immune system is deeply interconnected
with the endogenous 24-h oscillators of the circadian sys-
tem. Indeed, the connection between these two
physiological systems occurs at multiple levels and in both
directions. On one hand, various aspects of the immune
system show daily rhythms, which appear to be essential
for healthy immune maintenance and proper immune
response. On the other hand, immune responses cause
changes in circadian rhythms, disrupting their delicate
balance and manifesting in disease. Indeed, immune chal-
lenges cause various time-, gene-, and tissue-specific
effects on circadian-regulated factors. This article reviews
the possible mediators of the cross talk between the cir-
cadian clock and the immune system, in particular the
inflammatory pathways. The rhythmic expression of cyto-
kines and their receptors, as well as other rhythmically
regulated humoral factors such as glucocorticoids, mela-
tonin, leptin, or prostaglandins, could gate the effects of the
immune response on the circadian system. In addition,
systemic cues such as body temperature and neuronal
connections between the brain and peripheral tissues may
underlie the immune–circadian communication.
Keywords Circadian rhythm Clock gene Cytokine
Fever Innate immunity Inflammation
Distinct VEGF Functions During Bone Development
and Homeostasis
Yanqiu Liu • Bjorn R. Olsen
Received: 7 October 2013 / Accepted: 18 February 2014 / Published online: 4 April 2014
Ó L. Hirszfeld Institute of Immunology and Experimental Therapy, Wroclaw, Poland 2014
Abstract Vascular endothelial growth factor-A (VEGF)
is a key regulator of physiological hemangiogenesis during
development, postnatal growth, and homeostasis. It is well
known that VEGF is required for effective coupling of
angiogenesis to endochondral and membranous bone for-
mation during skeletal development. However, less well
known are the roles of VEGF in regulating the differenti-
ation and/or functions of skeletal cells such as
chondrocytes, osteoblasts, and osteoclasts. In this review,
we discuss some of these functions. During early skeletal
development, VEGF is important for the survival of
chondrocytes in the hypoxic regions of the cartilage models
of future bones, the vascularization of developing bones
and proliferation and differentiation of osteoblastic cells.
Postnatally, osteoblast-derived VEGF is critical for main-
taining bone homeostasis by stimulating the differentiation
of mesenchymal stem cells to osteoblasts and repressing
their differentiation to adipocytes. Recent data indicate that
these effects of VEGF on osteogenic/adipogenic stem cell
fates are based on an intracellular (intracrine) mechanism.
In contrast, osteoblast-derived VEGF is also known to
stimulate the differentiation of monocytes to osteoclasts by
a paracrine mechanism. Mice with VEGF-deficient osteo-
blastic lineage cells exhibit age-dependent loss of bone
mass and an increase in bone marrow fat. These changes
are similar to the changes associated with osteoporosis in
humans. Thus, a better understanding of the intracellular
mechanisms by which VEGF regulates osteoblastic and
adipogenic differentiation may lead to the identification of
new targets for therapies to prevent osteoporotic bone loss.
Keywords VEGF Mesenchymal stem cells
Ossification Osteoblast Adipocyte Differentiation
Helper T-Cell Differentiation in Graft-Versus-Host Disease
After Allogeneic Hematopoietic Stem Cell Transplantation
Jianing Fu • Jessica Heinrichs • Xue-Zhong Yu
Received: 23 September 2013 / Accepted: 27 January 2014 / Published online: 4 April 2014
Ó L. Hirszfeld Institute of Immunology and Experimental Therapy, Wroclaw, Poland 2014
Abstract Allogeneic hematopoietic stem cell transplan-
tation (allo-HSCT) is an effective therapeutic option for
many malignant diseases. However, the efficacy of allo-
HSCT is limited by the occurrence of destructive graft-
versus-host disease (GVHD). Since allogeneic T cells
are the driving force in the development of GVHD, their
activation, proliferation, and differentiation are key factors
to understanding GVHD pathogenesis. This review focuses
on one critical aspect: the differentiation and function of
helper T (Th) cells in acute GVHD. We first summarize
well-established subsets including Th1, Th2, Th17, and
T-regulatory cells; their flexibility, plasticity, and epige-
netic modification; and newly identified subsets including
Th9, Th22, and T follicular helper cells. Next, we exten-
sively discuss preclinical findings of Th-cell lineages in
GVHD: the networks of transcription factors involved in
differentiation, the cytokine and signaling requirements for
development, the reciprocal differentiation features, and
the regulation of microRNAs on T-cell differentiation.
Finally, we briefly summarize the recent findings on the
roles of T-cell subsets in clinical GVHD and ongoing
strategies to modify T-cell differentiation for controlling
GVHD in patients. We believe further exploration and
understanding of the immunobiology of T-cell differenti-
ation in GVHD will expand therapeutic options for the
continuing success of allo-HSCT.
Keywords Allo-HSCT GVL GVHD
T-cell differentiation
In vitro Correction of a Novel Splicing Alteration in the BTK Gene
by Using Antisense Morpholino Oligonucleotides
Natthakorn Rattanachartnarong • Siraprapa Tongkobpetch •
Pantipa Chatchatee • Tassalapa Daengsuwan • Chupong Ittiwut •
Kanya Suphapeetiporn • Vorasuk Shotelersuk
Received: 29 August 2013 / Accepted: 5 February 2014 / Published online: 23 March 2014
Ó L. Hirszfeld Institute of Immunology and Experimental Therapy, Wroclaw, Poland 2014
Abstract A novel sequence variant, c.240?109C[A, in
the Bruton’s tyrosine kinase (BTK) gene was identified in a
patient with X-linked agammaglobulinemia. This alteration
resulted in an incorporation of 106 nucleotides of BTK
intron 3 into its mRNA. Administration of the 25-mer
antisense morpholino oligonucleotide analog in the
patient’s cultured peripheral blood mononuclear cells was
able to restore correctly spliced BTK mRNA, a potential
treatment for X-linked agammaglobulinemia.
Keywords X-linked agammaglobulinemia BTK
Mis-splicing Antisense morpholino oligonucleotides
The Role of Serum C-Reactive Protein Measured
by High-Sensitive Method in Thyroid Disease
Agata Czarnywojtek • Maciej Owecki • Małgorzata Zgorzalewicz-Stachowiak • Kosma Wolin´ ski •
Ewelina Szczepanek-Parulska • Bartłomiej Budny • Ewa Florek • Joanna Waligo´rska-Stachura •
Izabela Miechowicz • Maciej Ba˛czyk • Nadia Sawicka • Sumit Dhir • Marek Ruchała
Received: 11 February 2013 / Accepted: 11 October 2013 / Published online: 4 May 2014
Ó The Author(s) 2014. This article is published with open access at Springerlink.com
Abstract The aim of this study was the evaluation of
serum C-reactive protein (CRP) concentration as a marker
of the inflammatory state in many different thyroid diseases
and its dependence on the stage and duration of disease.
We conducted a retrospective analysis of 444 randomly
selected patients with different kinds of thyroid disease
(106 men and 338 women, ranging 18–72 years of age;
mean 56.2 ± 5.0 years; median 52 years). Group 1 (G1)
comprised 250 patients with hyperthyroidism. Group 2
(G2) consisted of 72 euthyroid patients. Group 3 (G3)
consisted of 122 patients with hypothyroidism. Free T4,
free T3, and thyrotropin (TSH) levels were measured using
the electrochemiluminescent method. Human serum thy-
roglobulin autoantibodies (Tg-Abs), thyroperoxidase
autoantibodies (TPO-Abs), and autoantibodies against the
thyrotropin receptor (TSHR-Abs) levels were measured by
radioimmunoassay. The high-sensitive CRP (Hs-CRP)
level (reference range \3 mg/L) was determined with a
highly sensitive latex-based immunoassay. The mean value
of Hs-CRP in G1 was 3.6 ± 2.8 mg/L, in G2
2.5 ± 1.5 mg/L and in G3 5.9 ± 5.8 mg/L. Hs-CRP (in
mg/L) medians, interquartile and the total ranges in G1
were 3.0 (2.0 [0.1–21.0] 4.0); in G2: 2.3 [1.8 (0.2–9.2) 3.2];
and in G3: 4.3 [2.2 (0.3–31.5) 7.8]. We found statistically
significant differences (Kruskal–Wallis test) in serum Hs-
CRP values between G1 and G2 (P = 0.007), G1 and G3
(P = 0.001), G2 and G3 (P \ 0.001). In G1, statistically
significant correlation was confirmed between Hs-CRP and
Tg-Abs (r = –0.22, P = 0.0016), CRP and TPO-Abs
(r = –0.26, P \ 0.001), and also between Hs-CRP and
TSHR-Abs (r = –0.18, P = 0.02). In the remaining cases,
differences between Hs-CRP and TSH levels (r = –0.09,
P = 0.16) were not statistically significant. In G2, no sta-
tistically significant correlation was observed: Hs-CRP and
Tg-Abs (r = –0.18, P = 0.13), Hs-CRP and TPO-Abs
(r = –0.17, P = 0.15), Hs-CRP and TSH (r = 0.01,
P = 0.91), Hs-CRP and TSHR-Abs (r = –0.19,
P = 0.17). In G3, a statistically significant correlation was
confirmed between Hs-CRP and Tg-Abs (r = 0.22,
P = 0.012), Hs-CRP and TSH (r = –0.28, P = 0.001).
No statistically significant correlation was observed
between Hs-CRP and TPO-Abs (r = 0.20, P = 0.06) and
between Hs-CRP and TSHR-Abs (r = –0.23, P = 0.11).
Hs-CRP is increased in various types of hypothyroidism.
This is particularly relevant in postpartum thyroiditis and in
patients after radioiodine treatment. The impact of this
situation on human health requires further research, how-
ever, one might assume that some types of thyroid disease
may lead to systemic inflammatory reactions that are
reflected in elevated CRP levels.
Keywords Hs-CRP Thyroid disease
Hashimoto’s thyroiditis Graves’ disease Thyroid cancer
The MEK1/2–ERK1/2 Pathway is Activated in Chronic
Rhinosinusitis with Nasal Polyps
Robert Linke • Ralph Pries • Michael Ko¨nnecke •
Karl-Ludwig Bruchhage • Robert Bo¨scke •
Maximilian Gebhard • Barbara Wollenberg
Received: 19 February 2013 / Accepted: 11 December 2013 / Published online: 9 March 2014
Ó L. Hirszfeld Institute of Immunology and Experimental Therapy, Wroclaw, Poland 2014
Abstract Chronic rhinosinusitis with nasal polyps
(CRSwNP) is a common disease that has a considerable
impact on the quality of life. Alterations in signalling
pathways may contribute to the ongoing inflammation
and proliferation in CRSwNP. The MEK1/2–ERK1/2
pathway transmits signals from many extracellular mol-
ecules to regulate cellular processes. We examined tissue
samples from nasal polyps and the inferior turbinate of
patients with CRSwNP and the inferior turbinate from
subjects with healthy mucosa. The expressions of MEK1/
2, ERK1/2, and their active phosphorylated forms
pMEK1/2 and pERK1/2 were analysed using DNA
microarray, quantitative real-time PCR, protein array,
Western hybridisation, and immunohistochemistry. We
detected increased MEK1/2 protein expression in nasal
polyps compared to the inferior turbinates of patients
with CRSwNP or healthy mucosa. We also found a higher
amount of MEK1/2 in the inferior turbinates of patients
with CRSwNP compared to those with healthy mucosa.
Most importantly, we observed a significant increase in
the phosphorylation of MEK1/2 and ERK1/2 in nasal
polyps compared to both types of controls. We observed
activation of the MEK1/2–ERK1/2 pathway in nasal
polyps. Interestingly, we did not see the same activation
pattern in different tiers of the MEK1/2–ERK1/2 sig-
nalling cascade. One explanation for this result is that the
components enhance the complex MEK–ERK cascade in
a distinct manner, enabling a wide variety of functions.
The MEK1/2–ERK1/2 pathway appears to play a pivotal
role in the pathogenesis of CRSwNP.
Keywords Chronic rhinosinusitis ERK1/2 MEK1/2
Nasal polyp Phosphokinase
Double Transgenic Pigs with Combined Expression of Human
a1,2-Fucosyltransferase and a-Galactosidase Designed to Avoid
Hyperacute Xenograft Rejection
Joanna Zeyland • Anna Woz´niak • Barbara Gawron´ ska • Wojciech Juzwa •
Jacek Jura • Agnieszka Nowak • Ryszard Słomski • Zdzisław Smora˛g •
Marlena Szalata • Urszula Mazurek • Daniel Lipin´ ski
Received: 31 August 2013 / Accepted: 4 December 2013 / Published online: 20 February 2014
Ó The Author(s) 2014. This article is published with open access at Springerlink.com
Abstract Hyperacute rejection (HAR) depends on the
response of xenoreactive antibodies principally against
porcine a-Gal epitope. Methods eliminating HAR include
GGTA1 inactivation, regulation of the complement system
and modification of the oligosaccharide structure of surface
proteins in donor’s cells. Transgenic animals designed for
the purpose of xenotransplantation with single modification
do not display full reduction of the a-Gal epitope level,
which means that a accumulation of several modifications
in one transgenic individual is needed. The aim of the study
was to create a molecular and cytogenetic profile of a
double transgenic animal with a1,2-fucosyltransferase and
a-galactosidase expression. As a result of interbreeding of
an individual with a1,2-fucosyltransferase expression with
an individual with a-galactosidase expression 12 living
piglets were obtained. PCR revealed the pCMVFUT gene
construct was present in four individuals and pGAL-
GFPBsd in three, including one with a confirmed integra-
tion of both the gene constructs. Fluorescence in situ
hybridization confirmed the site of transgene integration,
which corresponded to the mapping site of the transgenes
which occurred in the parental generations. Karyotype
analysis did not show any changes in the structure or the
number of chromosomes (2n = 38, XX). As for the results
pertaining to the single transgenic individuals, expression
analysis demonstrated a high extent of a-Gal epitope level
reduction on the surface of cells, whereas human serum
cytotoxicity tests revealed the smallest decrease in lon-
gevity of cells in the obtained double transgenic individual
(4.35 %). The tests suggest that the co-expression of both
the transgenes leads to a considerable reduction of the a–
Gal antigen level on the surface of cells and a decrease of
xenotransplant immunogenicity.
Keywords a1,2-fucosyltransferase a-galactosidase
Hyperacute xenograft rejection Xenotransplantation
Hematopoietic Stem Cell Transplantation in Children
with Autoimmune Connective Tissue Diseases
Magdalena Witkowska • Elzbieta Smolewska •
Piotr Smolewski
Received: 17 July 2013 / Accepted: 15 November 2013 / Published online: 7 March 2014
Ó L. Hirszfeld Institute of Immunology and Experimental Therapy, Wroclaw, Poland 2014
Abstract Autoimmune connective tissue diseases
(ACTDs) are heterogeneous disorders associated with dif-
ferent manifestations, clinical course of disease and
prognosis among patients. Although recent advances in
understanding the pathogenesis have led to major progress
in target-oriented therapy, they still remain incurable.
Novel biological drugs, cellular therapy and hematopoietic
stem cell transplantation (HSCT) are real hopes for treat-
ment development in the future. The concept of both
autologous and allogeneic HSCT in children with autoim-
mune diseases is developing energetically since 1996,
when the first HSCT was performed. Nowadays, after
17 years of clinical experience, both types of HSCT remain
attractive and powerful salvage methods of treatment.
However, there are still many doubts and unclear issues,
which need further investigation. In the present review, we
provide an overview of the knowledge concerning actual
data on HSCT in a pediatric group of patients with different
ACTDs, focused on juvenile idiopathic arthritis, systemic
lupus erythematosus and systemic sclerosis.
Keywords Autoimmune connective tissue diseases
Children Treatment Hematopoietic
stem cell transplantation
Pregnancy-Induced Hypertension is Accompanied by Decreased
Number of Circulating Endothelial Cells and Circulating
Endothelial Progenitor Cells
Jerzy Heimrath • Maria Paprocka • Andrzej Czekanski •
Agata Ledwozyw • Aneta Kantor • Danuta Dus
Received: 11 July 2013 / Accepted: 11 December 2013 / Published online: 23 February 2014
Ó The Author(s) 2014. This article is published with open access at Springerlink.com
Abstract Maternal endothelial dysfunction is one of the
main features of pregnancy-induced hypertension (PIH). It
is generally accepted that circulating endothelial cells
(CECs) and endothelial progenitor cells (EPCs) reflect the
state of the endothelium, its injury and/or repair possibili-
ties. The objective of this study was to determine whether
the CECs and EPCs numbers in the circulation of women
with PIH reflect the presence of this pathology. Peripheral
blood cells of PIH and normotensive pregnant women were
labeled with specific monoclonal antibodies. For CECs
evaluation, samples were labeled with anti-CD31 and anti-
CD45 antibodies; for EPCs with anti-VEGFR2/KDR and
anti-CD34 antibodies. Cells were quantified by flow
cytometry. The levels of both CECs (CD31?, CD45–) and
EPCs (CD34?, VEGFR2/KDR?) in the peripheral blood of
women with PIH were significantly lower compared with
those of control pregnant women with normal blood pres-
sure level. Lowered accessibility of maternal CECs and
EPCs may diminish general regenerative potential of the
patient endothelia, contributing to PIH symptoms and to
the risk of subsequent coronary and arterial disease.
Keywords CECs EPCs Preeclampsia
Effect of Oral Administration Involving a Probiotic Strain
of Lactobacillus reuteri on Pro-Inflammatory Cytokine Response
in Patients with Chronic Periodontitis
Anna K. Szkaradkiewicz • Janina Stopa •
Tomasz M. Karpin´ ski
Received: 18 June 2013 / Accepted: 15 November 2013 / Published online: 9 February 2014
Ó The Author(s) 2014. This article is published with open access at Springerlink.com
Abstract This study aimed at evaluation of pro-inflam-
matory cytokine response (TNF-a, IL-1b and IL-17) in
patients with chronic periodontitis administered per os with
a probiotic strain of Lactobacillus reuteri. In the 38 adult
patients with moderate chronic periodontitis, professional
cleaning of teeth was performed. Two weeks after per-
forming the oral hygienization procedures, clinical
examination permitted to distinguish a group of 24 patients
(Group 1) in whom treatment with probiotic tablets con-
taining L. reuteri strain, producing hydrogen peroxide
(Prodentis), was conducted. In the remaining 14 patients,
no probiotic tablet treatment was applied (the control
group; Group 2). From all patients in two terms, gingival
crevicular fluid (GCF) was sampled from all periodontal
pockets. Estimation of TNF-a, IL-lb and IL-17 in GCF was
performed using the ELISA method. After completion of
the therapy with probiotic tablets, 18 (75 %) of the patients
of Group 1 have manifested a significant decrease in levels
of studied pro-inflammatory cytokines (TNF-a, IL-1b and
IL-17). In parallel, we have detected an improvement of
clinical indices [sulcus bleeding index (SBI), periodontal
probing depth (PPD), clinical attachment level (CAL)]. At
individuals of Group 2 levels of studies, pro-inflammatory
cytokines and clinical indices (SBI, PPD, CAL) were sig-
nificantly higher than in Group 1. Results obtained in this
study indicate that application of oral treatment with tablets
containing probiotic strain of L. reuteri induces in most
patients with chronic periodontitis a significant reduction
of pro-inflammatory cytokine response and improvement
of clinical parameters (SBI, PPD, CAL). Therefore, such an
effect may result in a reduced activity of the morbid
process.
Keywords Chronic periodontitis Lactobacillus
Cytokines Inflammation Immunomodulatory effect
Response to: Research Integrity: the Experience of a Doubting
Thomas
Pere Puigdome`nech
The Chemokine CX3CL1 (Fractalkine) and its Receptor
CX3CR1: Occurrence and Potential Role in Osteoarthritis
Piotr Wojdasiewicz • Łukasz A. Poniatowski • Andrzej Kotela •
Jarosław Deszczyn´ ski • Ireneusz Kotela • Dariusz Szukiewicz
Received: 17 September 2013 / Accepted: 4 December 2013 / Published online: 21 February 2014
Ó The Author(s) 2014. This article is published with open access at Springerlink.com
Abstract Chemokines are molecules able to induce
chemotaxis of monocytes, neutrophils, eosinophils, lym-
phocytes and fibroblasts. The complex chemokine acts in
many physiological and pathological phenomena,
including those occurring in the articular cartilage. To
date, chemokine CX3CL1 (fractalkine) is the only
member of the CX3C class of chemokines with well-
documented roles in endothelial cells. CX3CL1 is a
unique chemokine that combines properties of chemoat-
tractant and adhesion molecule. The main roles of
CX3CL1 include promotion of leukocyte binding and
adhesion as well as activation of the target cells. The
soluble chemokine domain of CX3CL1 is chemotactic
for T cells and monocytes. CX3CL1 acts via its receptor,
CX3CR1, which belongs to a family of G protein-cou-
pled receptors. Stimulation of CX3CR1 activates both
CX3CL1-dependent and integrin-dependent migrations of
cells with synergistically augmented adhesion. Genetic
polymorphisms of CX3CR1 may significantly modify the
biological roles of CX3CL1, especially in pathologic
conditions. Osteoarthritis (OA) is the most common joint
disease, affecting approximately 7–8 % of the general
population. Development of OA is largely driven by low-
grade local background inflammation involving chemo-
kines. The importance of CX3CL1/CX3CR1 signalling in
the pathophysiology of OA is still under investigation.
This paper, based on a review of the literature, updates
and summarises the current knowledge about CX3CL1/
CX3CR1 in OA and indicates possible interactions with
a potential for therapeutic targeting.
Keywords Chemokine CX3CL1 Fractalkine
Fractalkine receptor CX3CR1 Osteoarthritis
Hemophagocytic Syndrome in Children and Adults
Iwona Malinowska • Maciej Machaczka • Katarzyna Popko •
Alicja Siwicka • Małgorzata Salamonowicz •
Barbara Nasiłowska-Adamska
Received: 9 September 2013 / Accepted: 7 January 2014 / Published online: 9 February 2014
Ó The Author(s) 2014. This article is published with open access at Springerlink.com
Abstract Hemophagocytic syndrome, also known as
hemophagocytic lymphohistiocytosis (HLH), is a hetero-
genic syndrome, which leads to an acute, life-threatening
inflammatory reaction. HLH occurs both in children and
adults, and can be triggered by various inherited as well as
acquired factors. Depending on the etiology, HLH can be
divided into genetic (i.e., primary) and acquired (i.e., sec-
ondary) forms. Among genetic HLH forms, one can
distinguish between familial HLH and other genetically
conditioned forms of HLH. Acquired HLH can be typically
triggered by infections, autoimmune diseases, and malig-
nancies. The most common symptoms of HLH are
unremitting fever, splenomegaly, and peripheral blood
cytopenia. Some severely ill patients present with central
nervous system involvement. Laboratory tests reveal
hyperferritinemia (often [10,000 lg/L), increased serum
concentration of soluble receptor a for interleukin-2
([2,400 U/L), hypertriglyceridemia, hypofibrinogenemia,
coagulopathy, hyponatremia, hypoproteinemia, and ele-
vated liver transaminases and bilirubin. Prognosis in HLH
is very serious. Genetic HLH is always lethal if adequate
therapy is not administered. Similarly, severe acquired
cases often lead to death without appropriate treatment.
Since HLH can be encountered by various specialists in the
medical field, basic knowledge of this entity such as
diagnostic criteria and treatment should be familiar to all
physicians.
Keywords Hemophagocytic lymphohistiocytosis
Hemophagocytic syndrome Children Adults
The Anti-Tumor Activity of E1A and its Implications in Cancer
Therapy
Yi-Wen Chang • Mien-Chie Hung • Jen-Liang Su
Received: 29 May 2013 / Accepted: 17 January 2014 / Published online: 7 February 2014
Ó L. Hirszfeld Institute of Immunology and Experimental Therapy, Wroclaw, Poland 2014
Abstract The adenovirus type 5 E1A protein (E1A) plays
a critical role in anti-cancer gene therapy and has been
tested in clinical trials. The expression of E1A significantly
reduces tumorigenesis, promotes cell death, and inhibits
cancer cell mobility. Chemosensitization is one of the anti-
tumor effects of E1A, increasing in vitro and in vivo sen-
sitization of anti-cancer drugs, including cisplatin,
gemcitabine, etoposide, doxorubicin, paclitaxel, and tumor
necrosis factor-related apoptosis-inducing ligand and his-
tone deacetylase inhibitors in different types of cancer
cells. E1A also demonstrates anti-metastasis activity
through various molecular mechanisms such as the
repression of protease expression, suppression of HER2/
neu and downregulation of microRNA (miR-520h).
Moreover, E1A has been reported to reprogram transcrip-
tion in tumor cells and stabilize tumor suppressors such as
PP2A/C, p21 and p53. Because E1A plays a potentially
significant role in anti-tumor therapy, there exists an urgent
need to study the anti-cancer activities of E1A. This paper
presents a review of our current understanding of the
tumor-suppressive functions and molecular regulation of
E1A, as well as the potential clinical applications of E1A.
Keywords E1A Gene therapy Chemoresistance
Metastasis
Research Integrity: the Experience of a Doubting Thomas
Thomas P. Hettinger
Received: 18 November 2013 / Accepted: 10 January 2014 / Published online: 9 February 2014
Ó L. Hirszfeld Institute of Immunology and Experimental Therapy, Wroclaw, Poland 2014
Abstract The sensational ‘‘reactome array’’ paper pub-
lished in Science in 2009 was investigated in Spain by the
Ethics Committee of Consejo Superior de Investigaciones
Cientificas (CSIC) after Science issued an editorial
expression of concern. The paper was retracted in 2010
because of ‘‘skepticism’’ due to ‘‘errors’’ in chemistry. The
‘‘errors’’ were so profound that many readers expressed
doubt that they were really errors, but part of an elaborate
hoax. I conducted a forensic analysis of mass spectrometry
data in the paper’s Supporting Online Material (SOM) and
was able to prove that thousands of data values were in fact
fabricated. The SOM contains signatures of improper
extensive spreadsheet manipulations of incorrect atomic
and molecular mass values as well as impossibly repetitive
deviations of found molecular mass values from their
expected values. No evidence of real mass spectrometry
data was detected. Both CSIC and Science have been
content to retract the paper without acknowledging the
fabrications or assigning responsibility for them. Neither
CSIC nor Science has expressed interest in having an
independent investigation determining how the paper came
to be written, reviewed and published. Their weak response
to this episode is a daunting signal that there is an
impending crisis in research integrity and science
journalism.
Keywords Ethics Forensic analysis Fraud
Reactome array Research misconduct
Science journalism
Effects of Resveratrol on the Expression and DNA Methylation
of Cytokine Genes in Diabetic Rat Aortas
Xu-dan Lou • Hai-dong Wang • Shi-jin Xia •
Sven Skog • Jiao Sun
Received: 3 December 2012 / Accepted: 30 July 2013 / Published online: 5 February 2014
Ó L. Hirszfeld Institute of Immunology and Experimental Therapy, Wroclaw, Poland 2014
Abstract This paper studies the expression of proin-
flammatory cytokines such as IL-1b, IL-6, TNF-a, and
IFN-c and anti-inflammatory cytokines such as IL-10 in
diabetic rat aortas, the effects of resveratrol on these
cytokines, and the potential epigenetic mechanisms
involved. The experiment was performed on rats divided
into four groups: normal group (NC), normal interventional
group (NB), diabetic group (DM), and diabetic interven-
tional group (DB). The NB and DB groups were treated
with resveratrol. After more than 3 months, the rats’ aortas
were removed and analyzed for cytokines by using
immunohistochemistry, Western blotting, real-time PCR,
and methylation-specific PCR. Histological localization of
these cytokines was mainly found in the arterial intima of
diabetic rats. The protein and mRNA expression levels of
IL-1b, IL-6, TNF-a, and IFN-c were significantly higher in
the DM group than in the NC group (p \ 0.05), whereas in
the resveratrol-treated groups (NB and DB), the levels were
relatively lower than those in the corresponding groups.
The DM group showed reduced levels of DNA methylation
at the specific cytosine phosphate guanosine sites of IL-1b,
IL-6, TNF-a, and IFN-c, relative to those in the NC group
(p \ 0.01), and these levels were increased by resveratrol.
In contrast, IL-10 was dramatically methylated and showed
decreased expression in response to high glucose, and
resveratrol reversed this effect. These results demonstrate
that the inflammatory response is involved in diabetic
macroangiopathy. Resveratrol inhibits the expression of
proinflammatory cytokines and thus may have a protective
effect on the aorta in hyperglycemia. Thus, DNA methyl-
ation, an epigenetic gene silencing signal, may be
responsible for these two phenomena.
Keywords Inflammatory cytokines Resveratrol
DNA methylation
Exhaled IL-8 in Systemic Lupus Erythematosus
with and without Pulmonary Fibrosis
Agnieszka Nielepkowicz-Goz´dzin´ ska • Wojciech Fendler • Ewa Robak •
Lilianna Kulczycka-Siennicka • Paweł Go´rski • Tadeusz Pietras •
Ewa Brzezian´ ska • Adam Antczak
Received: 17 December 2012 / Accepted: 9 October 2013 / Published online: 4 February 2014
Ó The Author(s) 2014. This article is published with open access at Springerlink.com
Abstract The purpose of this study is to evaluate the
relationship between the concentration of interleukin-8 (IL-
8) in exhaled breath condensate (EBC) and bronchoalve-
olar lavage fluid (BALF) with the disease activity score and
pulmonary function of systemic lupus erythematosus (SLE)
patients with and without pulmonary fibrosis. Thirty-four
SLE patients and 31 healthy controls were enrolled and
evaluated using high-resolution computed tomography
(HRCT), pulmonary function tests, systemic lupus activity
measure (SLAM), assessing BALF and EBC. IL-8 levels in
BALF and EBC samples were measured with an enzyme-
immunosorbent assay kit. The mean (±SEM) IL-8 con-
centrations in BALF and EBC were higher in SLE patients
compared to healthy controls (34.84 ± 95.0 vs.
7.65 ± 21.22 pg/ml, p \ 0.001; 3.82 ± 0.52 pg/m vs.
1.7 ± 1.7 pg/ml, p \ 0.001, respectively). SLE patients
had increased percentage of neutrophils in BALF when
compared with control group (1.00 ± 5.99 vs.
0.00 ± 0.56 %, p = 0.0003). Pulmonary fibrosis in HRCT
was found in 50 % of SLE patients. The disease activity
scored by SLAM was significantly higher and total lung
capacity was significantly lower in SLE patients with
pulmonary fibrosis (8.00 ± 3.17 vs. 6.00 ± 2.31,
p = 0.01; 88.00 ± 28.29 vs. 112.00 ± 21.08 % predicted,
p = 0.01, respectively). In SLE patients with pulmonary
fibrosis, correlations were found between SLAM and IL-8
concentration in BALF, forced expiratory volume in 1 s
and forced vital capacity (r = 0.65, p = 0.006; r = –0.53,
p = 0.035; r = –0.67, p = 0.006, respectively). Our
results indicate that IL-8 plays an important role in the
pathogenesis of SLE. An increased concentration of IL-8
according to BALF could be considered as a useful bio-
marker of SLE activity and pulmonary fibrosis in SLE.
Keywords Systemic lupus erythematosus
Interleukin-8 Exhaled breath condensate
Bronchoalveolar lavage fluid Pulmonary fibrosis
Call for a Dedicated European Legal Framework
for Bacteriophage Therapy
Gilbert Verbeken • Jean-Paul Pirnay •
Rob Lavigne • Serge Jennes • Daniel De Vos •
Minne Casteels • Isabelle Huys
Received: 8 July 2013 / Accepted: 6 November 2013 / Published online: 6 February 2014
Ó The Author(s) 2014. This article is published with open access at Springerlink.com
Abstract The worldwide emergence of antibiotic resis-
tances and the drying up of the antibiotic pipeline have
spurred a search for alternative or complementary antibac-
terial therapies. Bacteriophages are bacterial viruses that
have been used for almost a century to combat bacterial
infections, particularly in Poland and the former Soviet
Union. The antibiotic crisis has triggered a renewed clinical
and agricultural interest in bacteriophages. This, combined
with new scientific insights, has pushed bacteriophages to
the forefront of the search for new approaches to fighting
bacterial infections. But before bacteriophage therapy can
be introduced into clinical practice in the European Union,
several challenges must be overcome. One of these is the
conceptualization and classification of bacteriophage ther-
apy itself and the extent to which it constitutes a human
medicinal product regulated under the European Human
Code for Medicines (Directive 2001/83/EC). Can thera-
peutic products containing natural bacteriophages be
categorized under the current European regulatory frame-
work, or should this framework be adapted? Various actors
in the field have discussed the need for an adapted (or
entirely new) regulatory framework for the reintroduction
of bacteriophage therapy in Europe. This led to the identi-
fication of several characteristics specific to natural
bacteriophages that should be taken into consideration by
regulators when evaluating bacteriophage therapy. One
important consideration is whether bacteriophage therapy
development occurs on an industrial scale or a hospital-
based, patient-specific scale. More suitable regulatory
standards may create opportunities to improve insights into
this promising therapeutic approach. In light of this, we
argue for the creation of a new, dedicated European regu-
latory framework for bacteriophage therapy.
Keywords Bacteriophage Therapy Human
European Regulatory Legal Legislation
D-K6L9 Peptide Combination with IL-12 Inhibits the Recurrence
of Tumors in Mice
Tomasz Cichon´ • Ryszard Smolarczyk •
Sybilla Matuszczak • Magdalena Barczyk •
Magdalena Jarosz • Stanisław Szala
Received: 28 September 2012 / Accepted: 11 October 2013 / Published online: 2 February 2014
Ó The Author(s) 2014. This article is published with open access at Springerlink.com
Abstract D-K6L9 peptide is bound by phosphatidylserine
and induces necrosis in cancer cells. In our therapeutic
experience, this peptide, when administered directly into
B16-F10 murine melanoma tumors, inhibited their growth.
Cessation of therapy results, however, in tumor relapse. We
aimed at developing a combined therapy involving D-K6L9
and additional factors that would yield complete elimina-
tion of tumor cells in experimental animals. To this
purpose, we employed glycyrrhizin, an inhibitor of
HMGB1 protein, BP1 peptide and interleukin (IL)-12.
Glycyrrhizin or BP1, when combined with D-K6L9,
inhibits growth of primary tumors only during the period of
their administration. A long-term tumor growth inhibitory
effect was obtained only in combining D-K6L9 with IL-12.
At 2 months following therapy cessation, 60 % of animals
were alive. Prolonged survival was noted in mice bearing
B16-F10 tumors as well as in mice bearing C26 colon
carcinoma tumors.
Keywords Peptide D-K6L9 Peptide BP1 Glycyrrhizin
IL-12 Anticancer therapy