XII European Symposium on Platelet and Granulocyte Immunobiology
Immune Regulation by Sphingosine 1-Phosphate and Its Receptors
Constantin Bode • Markus H. Gra¨ler
Abstract It is well established that the lysophospholipid and signalling molecule sphingosine 1-phosphate (S1P) has many important functions in immune surveillance. S1P is produced from sphingosine by two distinct sphingosine kinases, SphK1 and SphK2, and acts as an intracellular messenger and as an extracellular ligand of five G proteincoupled cell surface receptors designated S1P1–S1P5. S1P not only regulates peripheral lymphocyte circulation, but also influences their differentiation, activation, infiltration, and local positioning. The therapeutic value of modulating S1P metabolism and S1P receptor function is currently tested in clinical trials and holds great promise for treatment of different autoimmune diseases. Despite its obvious contribution to immune regulation, the analysis of S1P is still challenging. A major obstacle is the difficulty to analyze S1P locally in tissues and within cells due to its high metabolic turnover and the limited resolution of current analytical techniques like liquid chromatography and mass spectrometry. This review focuses on recent advancements to our understanding how different sources of S1P contribute to immune function, and how changes in production, secretion, and degradation of S1P can influence immune responses.
Keywords Lymphocyte egress Sphingosine kinase S1P-lyase Sphingolipid metabolism Lymphopenia FTY720
Acetylcholine Receptor-Induced Experimental Myasthenia Gravis: What Have We Learned from Animal Models After Three Decades?
Fulvio Baggi • Carlo Antozzi • Chiara Toscani • Chiara Cordiglieri
Abstract Myasthenia gravis (MG) is an autoimmune disease caused by an immunological response against the acetylcholine receptor (AChR) at the neuromuscular junction. Anti-AChR antibodies induce degradation of the receptor, activation of complement cascade and destruction of the post-synaptic membrane, resulting in a functional reduction of AChR availability. The pathophysiological role of autoantibodies (auto-Abs) and T helper lymphocytes has been studied in the experimental autoimmune MG (EAMG) models. EAMG models have been employed to investigate the factors involved in the development of MG and to suggest new therapies aimed to preventing or modulating the ongoing disease. EAMG can be induced in susceptible mouse and rat strains, which develop clinical symptoms such as muscular weakness and fatigability, mimicking the human disease. Two major types of EAMG can be induced, passive and active EAMG. Passive transfer MG models, involving the injection of auto-Abs, are helpful for studying the role of complement molecules and their regulatory proteins, which can prevent neuromuscular junction degradation. Active models, induced by immunization, are employed for the analysis of antigen-specific immune responses and their modulation in order to improve disease progression. In this review, we will concentrate on the main pathogenic mechanisms of MG, focusing on recent findings on EAMG experimental models.
Keywords Myasthenia gravis Autoimmunity Neuroimmunology Experimental model
Immunotherapy of Lung Adenocarcinoma Patient
with Peptide-Pulsed Dendritic Cells: a Case Report
Kamila Wojas-Krawczyk • Paweł Krawczyk •
Jarosław Buczkowski • Anna Walkowska • Olga Jankowska •
El_zbieta Czekajska-Chehab • Janusz Milanowski
Received: 6 May 2011 / Accepted: 14 July 2011 / Published online: 6 December 2011
Ó L. Hirszfeld Institute of Immunology and Experimental Therapy, Wroclaw, Poland 2011
Abstract Immunotherapy with ex vivo generated den-
dritic cells (DCs) is reported to be of low toxicity and of
diverse effectiveness in cancer treatment. The synthetic
antigens are frequently used for immunotherapy especially
for patients with stable disease after prior treatment. We
described the effect of peptide-loaded DCs-based immu-
notherapy on patient with recurrent surgically resected
adenocarcinoma with bronchoalveolar feature with co-
existing of Takayasu arteritis and chronic hepatitis B. In
January 2010, 61-year-old patient received subcutaneously
four bi-weekly vaccinations of DCs loaded with MUC1 and
MAGE-3 epitopes. Additionally, he received three bi-
weekly booster vaccinations after 7 months from the first
course of immunotherapy. Delayed-type hypersensitivity
test was positive only for MAGE-3 antigen. The evidence
expansion of MAGE-3-specific CD8? cells after first vac-
cination and after third vaccination during boosters
injections was observed (from 0.08% before vaccination to
0.5% after first vaccination; from 0.05% before booster
vaccination to 0.24% after third injection). Computed
tomography scans performed after first course and after
booster course of vaccination until April 2011 did not
shown any presence of lung tumour or metastases. Based
on clinical factors (no completed wedge-resection and
recurrent character of cancer) as well as on the presence of
the tumour-antigen-specific immunological response, we
could speculated that immunotherapy prolonged disease
free-survival in our patient. Over 16 months from first
vaccination, the patient remains without symptoms of
cancer.
Keywords Dendritic cells Immunotherapy
Adenocarcinoma Synthetic peptide
Neutrophil Myeloperoxidase: Soldier and Statesman
Zofia Prokopowicz • Janusz Marcinkiewicz •
David R. Katz • Benjamin M. Chain
Received: 20 May 2011 / Accepted: 5 October 2011 / Published online: 6 December 2011
Ó L. Hirszfeld Institute of Immunology and Experimental Therapy, Wroclaw, Poland 2011
Abstract Myeloperoxidase (MPO) is a major protein
constituent of the primary granules of vertebrate neutro-
phils. It catalyses the hydrogen peroxide-mediated
oxidation of halide ions to hypohalous acids, especially
HOCl. These reactive oxygen species can participate in a
variety of secondary reactions, leading to modifications of
amino acids and many types of biological macromolecules.
The classic paradigm views MPO as a component of the
phagocyte oxygen-dependent intracellular microbicidal
system, and thus an important arm of the effector phase of
innate immune responses. However, the limited immuno-
deficiency associated with lack of MPO in mouse and
human models has challenged this paradigm. In this review
we examine more recent information on the interaction
between MPO, its bioreactive reaction products, and tar-
gets within the inflammatory microenvironment. We
propose that two assumptions of the current model may
require revisiting. First, many important targets of MPO
modification are extracellular, rather than present only
within the phagolysosome, such as various components of
neutrophil extracellular traps. Second, we suggest that the
pro-inflammatory pathological role of MPO may be a
particular feature of chronic inflammation. In the physio-
logical setting of acute neutrophil-mediated inflammation
MPO may also form part of a negative feedback loop
which down-regulates inflammation, limits tissue damage,
and facilitates the switch from innate to adaptive immunity.
This different perspective on this well-studied enzyme may
usefully inform further research into its function in health
and disease.
Keywords Myeloperoxidase Hypochlorous acid
Neutrophils Acute and chronic inflammation
Molecular Mechanisms and Pathological Consequences
of Endotoxin Tolerance and Priming
Matthew Morris • Liwu Li
Received: 23 June 2011 / Accepted: 27 September 2011 / Published online: 6 December 2011
Ó L. Hirszfeld Institute of Immunology and Experimental Therapy, Wroclaw, Poland 2011
Abstract Lipopolysaccharide (LPS), a component of
Gram-negative bacteria, is a potent inflammatory stimu-
lant, with high doses due to disseminated bacterial
infection resulting in systemic inflammatory response
syndrome and death. Lower doses can induce a state of
tolerance to subsequent toxic doses of LPS, but extremely
low doses have an opposite effect, priming the immune
system for an even more violent response to subsequent
challenge. A substantial body of research exists on the
phenomenon of endotoxin tolerance, which appears to be a
state of generalized dampening of inflammatory pathways.
Comparatively little is known about the mechanisms or
indeed the phenomenon of priming, particularly regarding
the shift from a priming to a tolerizing response. Our aim is
to review recent findings in the field of the inflammatory
response to endotoxin, with a focus on highlighting the
gaps in current understanding and attempting to reconcile
the competing tolerance and priming phenomena.
Keywords Endotoxin Tolerance Priming Sepsis
Inflammatory diseases
Lysophosphatidylcholines: Bioactive Lipids Generated During
Storage of Blood Components
Krystyna Mas´lanka • Gabriela Smolen´ ska-Sym •
Halina Michur • Agnieszka Wro´bel • El_zbieta Lachert •
Ewa Brojer
Received: 15 April 2011 / Accepted: 12 August 2011 / Published online: 14 December 2011
Ó L. Hirszfeld Institute of Immunology and Experimental Therapy, Wroclaw, Poland 2011
Abstract Transfusion-related acute lung injury (TRALI) is
suggested to be a ‘‘two hit’’ event, resulting from priming
and activation of pulmonary neutrophils. It is known that
neutrophil activation may result from infusion of lysophos-
phatidylcholines (LysoPCs) accumulated during storage of
blood components. The aim of our study was to verify
whether the LysoPCs are released into the storage medium of
blood components. We measured the LysoPCs concentration
in the supernatants from stored apheresis platelet concen-
trates (PLTs), packed non-leukoreduced red blood cell
concentrates (RBCs), leukoreduced red blood cell concen-
trates (L-RBCs), fresh frozen plasma (FFP) and donor
plasma (control). Lipids were separated on high-performance
thin-layer chromatography, detected by primulin spray and
quantified by photodensitometric scanning. The LysoPCs
concentration in donor plasma was similar to that in FFP.
During storage the LysoPCs content in PLTs increased
almost two-fold as compared to the fresh isolated platelets. In
RBCs and L-RBCs the LysoPC level was very low or below
detection limit and did not increase throughout the storage
period. According to our observations bioactive LysoPCs
may be considered a neutrophil-activating factor only fol-
lowing PLT transfusions but not RBCs transfusions.
Keywords Bioactive lipids Lysophosphatidylcholines
Storage of blood components TRALI
Neutrophil Function and Apoptosis in Patients with Chronic
Hepatitis C Treated with Pegylated Interferon a and Ribavirin
El_zbieta Jabłonowska • Kamila Wo´jcik •
Barbara Kur • Przemysław Lewkowicz •
Marek Nocun´
Received: 30 January 2011 / Accepted: 1 July 2011 / Published online: 8 December 2011
Ó L. Hirszfeld Institute of Immunology and Experimental Therapy, Wroclaw, Poland 2011
Abstract The role of neutrophils in the pathogenesis of
chronic hepatitis C as well as the effect of pegylated inter-
feron a (PEG-IFN-a) and ribavirin treatment on neutrophil
function is not precisely known. The study included 32
patients with CCH aged between 19 and 58 years (mean
33.5 years). Before and after 12 weeks of treatment with
Peg-IFN-a and ribavirin, intracellular reactive oxygen spe-
cies (ROS) level, expression of adhesion molecules CD11b/
MAC-1, CD16, CD18 and CD62L on neutrophils, as well as
apoptosis and necrosis of these cells were analyzed with the
use of flow cytometry. During antiviral therapy, a statisti-
cally significant decrease of mean fluorescence intensity for
CD16 high and CD62 and increase for CD11b/MAC-1 along
with the increased apoptosis and decreased necrosis of
neutrophils were observed. After 12 weeks of treatment,
intracellular ROS production by unstimulated neutrophils
did not change, but after stimulation with phorbol 12-myr-
istate 13-acetate, statistically significant increase of ROS
level was observed. During PEG-IFN-a and ribavirin treat-
ment, activation of neutrophil function and increased ROS
production were reported, which possibly resulted in
accelerated apoptosis of these cells.
Keywords HCV IFN Neutrophils Apoptosis
Allogeneic Hematopoietic Cell Transplantation
for Myelodysplastic Syndrome: Current Status
H. Joachim Deeg • Matthias Bartenstein
Received: 11 May 2011 / Accepted: 28 October 2011 / Published online: 6 December 2011
Ó L. Hirszfeld Institute of Immunology and Experimental Therapy, Wroclaw, Poland 2011
Abstract Hematopoietic cell transplantation (HCT) offers
potentially curative therapy for patients with myelodysplastic
syndrome (MDS). However, as the majority of patients with
MDS is in the 7th or 8th decade of life, only few of these
patients were transplanted following high-dose conditioning
regimens. The development of reduced-intensity conditioning
has allowed to apply HCT also to older patients and those with
clinically relevant comorbid conditions. Dependent upon
disease status and the type of clonal chromosomal abnor-
malities present at the time of HCT, some 25–75% of patients
will be cured of their disease and survive long term. Recent
results with HLA-matched unrelated donors are comparable
to those with HLA genotypically identical siblings. The
increasing use of cord blood and HLA-haploidentical donors
is expected to make HCT available to a growing number of
patients. However, post-transplant relapse and graft-versus-
host disease remain problems requiring further instigations.
Keywords Transplantation for MDS Age
Comorbidities Conditioning intensity Relapse
Archivum Immunologiae et Therapiae Experimentalis in Its 60th Year of Publication (1953–2012)
Angiogenesis in Oral Lichen Planus: An In Vivo
and Immunohistological Evaluation
Giuseppe Alessandro Scardina • Alessia Ruggieri •
Emiliano Maresi • Pietro Messina
Received: 6 July 2010 / Accepted: 21 February 2011 / Published online: 1 October 2011
Ó L. Hirszfeld Institute of Immunology and Experimental Therapy, Wroclaw, Poland 2011
Abstract Oral lichen planus (OLP) is an autoimmune
disease with an inflammatory pathogenesis. The angioge-
netic phenomenon is a mechanism at the base of the
pathogenesis of chronic inflammatory processes. The aim
of this research is to evaluate the angiogenetic phenome-
non, comparing an in vitro method with an in vivo one.
Thirty OLP patients and 30 healthy subjects were enrolled
in the study. Immunohistochemical analysis of the vascu-
lar-endothelial growth factor (VEGF) and vascular-
endothelial adhesion molecules were carried out by the
means of primary antibodies and anti-CD34, anti-VEGF,
anti-CD106 antigen (VCAM-1) and anti-CD54 antigen
(ICAM-1). Capillary density and others capillaroscopic
parameters were tested in vivo using oral videocapillaros-
copy. The results reveal the presence of a significant
angiogenesis in OLP patients through the immunoexpres-
sion of VEGF, CD34, CD106 and CD54 (p \ 0.001).
Capillaroscopic analysis demonstrates significant value for
the following parameters: density, tortuosity, loop diame-
ter, afferent and efferent capillary loop diameter. The in
vivo and in vitro investigation in OLP reveals a significant
angiogenesis.
Keywords Angiogenesis Videocapillaroscopy
VEGF Immunohistochemistry
Beclin 1 is Involved in Regulation of Apoptosis and Autophagy
During Replication of Ectromelia Virus in Permissive L929 Cells
Lech Martyniszyn • Lidia Szulc • Anna Boratyn´ska •
Marek G. Niemiałtowski
Received: 21 December 2010 / Accepted: 25 July 2011 / Published online: 5 October 2011
Ó L. Hirszfeld Institute of Immunology and Experimental Therapy, Wroclaw, Poland 2011
Abstract Several reports have brought to light new and
interesting findings on the involvement of autophagy and
apoptosis in pathogenesis of viral and bacterial diseases, as
well as presentation of foreign antigens. Our model studies
focused on the involvement of apoptosis during replication
of highly virulent Moscow strain of ectromelia virus
(ECTV-MOS). Here, we show evidence that autophagy is
induced during mousepox replication in a cell line. Fluo-
rescence microscopy revealed increase of LC3
(microtubule-associated protein 1 light chain 3) aggrega-
tion in infected as opposed to non-infected control L929
cells. Furthermore, Western blot analysis showed that
replication of ECTV-MOS in L929 cells led to the increase
in LC3-II (marker of autophagic activity) expression.
Beclin 1 strongly colocalized with extranuclear viral rep-
lication centers in infected cells, whereas expression of
Bcl-2 decreased in those centers as shown by fluorescence
microscopy. Loss of Beclin 1-Bcl-2 interaction may lead to
autophagy in virus-infected L929 cells. To assess if Beclin
1 has a role in regulation of apoptosis during ECTV-MOS
infection, we used small interfering RNA directed against
beclin 1 following infection. Early and late apoptotic cells
were analyzed by flow cytometry after AnnexinV and
propidium iodide staining. Silencing of beclin 1 resulted in
decreased percentage of early and late apoptotic cells in the
late stage of ECTV-MOS infection in L929 cells. We
conclude that Beclin 1 plays an important role in regulation
of both, autophagy and apoptosis, during ECTV-MOS
replication in L929 permissive cells.
Keywords Ectromelia (mousepox) virus Autophagy
Apoptosis
Disseminated Adenovirus Disease in Immunocompromised
Patient Successfully Treated with Oral Ribavirin: A Case Report
Ewa El_zbieta Ulrych • Tomasz Dziecia˛tkowski • Maciej Przybylski •
Dorota Zdun´ czyk • Piotr Boguradzki • Tigran Torosian •
Anna Waszczuk-Gajda • Sylwia Rynans • Marta Wro´blewska •
Wiesław Wiktor Je˛drzejczak • Gra_zyna Młynarczyk
Received: 17 January 2011 / Accepted: 28 June 2011 / Published online: 1 October 2011
Ó L. Hirszfeld Institute of Immunology and Experimental Therapy, Wroclaw, Poland 2011
Abstract In patients with immunological disorders, ade-
novirus infections are associated with significant rates of
morbidity and mortality. Only few hematological units use
molecular virological methods, such as polymerase chain
reaction, for surveillance of adenovirus infection, and
treatment strategies have never been evaluated in multi-
center clinical trials. This report describes the detection and
treatment of human adenovirus (HAdVs) disseminated
disease in the case of a 46-year-old immunocompromised
female having myelodysplastic syndrome with refractory
cytopenia with multilineage dysplasia: International Prog-
nostic Scoring System 1. Serum and urine samples were
tested for the presence of adenoviral DNA using the
quantitative real-time polymerase chain reaction (PCR)
assay. For additional confirmation, sequencing of PCR
products was also performed. With real-time PCR, we
detected HAdV DNA in both serum and urine samples. The
viral level constantly decreased with applied oral ribavirin
therapy. As the result of sequencing, HAdVs type 11 was
determined. Surveillance of adenovirus by real-time PCR is
useful in detecting and monitoring disseminated HAdV
infection; it is a potential standard diagnostic approach that
could assist clinicians to decide whether antiviral therapy
ought to be administered.
Keywords Adenovirus infection Immunosuppression
Myelodysplastic syndrome Real-time PCR
The Transcription Factor PU.1 is a Critical Regulator
of Cellular Communication in the Immune System
Shereen A. Turkistany • Rodney P. DeKoter
Received: 28 April 2011 / Accepted: 25 August 2011 / Published online: 5 October 2011
Ó L. Hirszfeld Institute of Immunology and Experimental Therapy, Wroclaw, Poland 2011
Abstract PU.1 is an E26 transformation-specific family
transcription factor that is required for development of the
immune system. PU.1 functions at both early and late
stages of lymphoid and myeloid differentiation. At least
110 direct target genes of PU.1 have been identified since
its discovery in 1988. We used the published literature to
determine if aspects of PU.1 function can be inferred from
the identity of target genes that are directly activated. This
analysis revealed that 61% of described PU.1 target genes
encode extracellular proteins or transmembrane proteins,
most of which are involved in cellular communication. The
genes activated by PU.1 can be grouped into pathways
based on function. Specific examples of cellular commu-
nication pathways regulated by PU.1 include (1) antibodies
and antibody receptors, (2) cytokines and cytokine recep-
tors regulating leukocyte growth and development, and
(3) cytokines and cytokine receptors regulating inflamma-
tion. As a consequence of mutation or repression of the
gene encoding PU.1, hematopoietic progenitors may be
generated but there is a ‘‘failure to thrive’’ because they
cannot interact with their environment. The loss of cellular
communication caused by reduced PU.1 levels can lead to
leukemia. In summary, PU.1 is a critical regulator of cel-
lular communication in the immune system.
Keywords Immune development Transcription factor
PU.1 Cytokine Antibody Leukemia
Complement in Cancer and Cancer Immunotherapy
Martin Kolev • Laurence Towner • Rossen Donev
Received: 5 April 2011 / Accepted: 7 June 2011 / Published online: 30 September 2011
Ó L. Hirszfeld Institute of Immunology and Experimental Therapy, Wroclaw, Poland 2011
Abstract Recently, there has been an increase of interest
in the use of biological or immune-based therapies for
patients with malignancies. This has been informed by the
deeper understanding of the crosstalk between the host
immune system and malignant tumours, as well as the
potential advantages of immunotherapy—high specificity
and less toxicity compared to standard approaches. The
particular emphasis of this article is on the role of the
complement system in tumour growth and antibody-based
cancer immunotherapy. The functional consequences from
overexpression of complement regulators by tumours and
the development of strategies for overcoming this are dis-
cussed in detail. This review discusses these issues with a
view to inspiring the development of new agents that could
be useful for the treatment of cancer.
Keywords Immunotherapy Cancer Complement
Complement regulators
Complement-dependent cytotoxicity (CDC)
Antibody-dependent cell-mediated cytotoxicity (ADCC)
NK Cells Prevalence, Subsets and Function in Viral Hepatitis C
Jan _Zeromski • Iwona Mozer-Lisewska •
Mariusz Kaczmarek • Arleta Kowala-Piaskowska •
Jan Sikora
Received: 18 April 2011 / Accepted: 4 July 2011 / Published online: 5 October 2011
Ó L. Hirszfeld Institute of Immunology and Experimental Therapy, Wroclaw, Poland 2011
Abstract Innate immunity appears to play an important
role in the pathogenesis of viral hepatitis C. Among various
cell subsets of this immunity natural killer (NK) cells
raised particular interest. These cells are abundant in liver,
possess significant cytotoxic potential and show links with
adaptive immunity. They play important role, particularly
in the acute phase of viral infections, including hepatitis
C. They exhibit various types of receptors, either inhibitory
or activating, that are able to react with distinct ligands on
infected cells. Homozygosity of some receptors, namely
KIR2DL3 reacting with recipient HLA-C1 antigens is a
herald of good prognosis in hepatitis C virus (HCV)
infection. In the early stage of the latter, both the preva-
lence and the cytotoxicity of NK cells are increased. Their
inhibitory receptors are down regulated whereas activating
ones are up regulated. Interferon-c secreted by NK56?bright
NK cells has a direct cytotoxic effect on infected hepato-
cytes. In contrast, in the chronic phase of HCV liver
disease both, the prevalence and function of NK cells are
impaired. Nevertheless, their cytotoxicity contributes to
liver injury. Cells show change in the polarization profile
from NK1 to NK2, manifested by secretion of immuno-
suppressive cytokines. Some HCV peptides are inhibitory
for NK cells leading to the reduction of their antiviral
activity. The unwanted effects of HCV peptides can be at
least partly reversed by the antiviral therapy.
Keywords NK cells Viral hepatitis C
NK receptors NK cytotoxicity Antiviral therapy
HCV-mediated NK immunosuppression
Oxidative and Nitrosative Stress on Phagocytes’ Function:
from Effective Defense to Immunity Evasion Mechanisms
Carlos K. B. Ferrari • Paula C. S. Souto •
Eduardo L. Franc¸a • Adenilda C. Honorio-Franc¸a
Received: 24 March 2011 / Accepted: 27 June 2011 / Published online: 5 October 2011
Ó L. Hirszfeld Institute of Immunology and Experimental Therapy, Wroclaw, Poland 2011
Abstract Although oxygen, nitrogen, and chlorine reac-
tive species have been associated with disease pathogenesis,
their partial absence is very harmful to the body’s innate
immune defense. Lacking of adequate release of free radi-
cals from activated phagocytes is related to impaired ability
on fungi, bacteria, and protozoa killing. We constructed an
updated conceptual landmark regarding the paramount role
of free radicals in phagocyte defense systems (phagocyte
oxidase, myeloperoxidase, and nitric oxide/peroxynitrite
system) on natural immunity. Diverse fungal, bacterial and
protozoal pathogens evade the phagocytes’ oxidative/
nitrosative burst though antioxidant genes, enzymes and
proteins. The most important evasion mechanisms were
also described and discussed. These interconnected systems
were reviewed and discussed on the basis of knowledge
from relevant research groups around the globe. Phagocyte-
derived free radicals are essential to destroy important
human pathogens during the course of innate immunity.
Keywords Phagocyte oxidase Macrophage
Myeloperoxidase Nitric oxide Peroxynitrite
Malaria Candida spp. Mycobacterium tuberculosis
Trypanosoma cruzi
Islet Transplantation: Factors in Short-Term Islet Survival
Natasha A. Deters • Rebecca A. Stokes •
Jenny E. Gunton
Received: 28 January 2011 / Accepted: 25 May 2011 / Published online: 8 October 2011
Ó L. Hirszfeld Institute of Immunology and Experimental Therapy, Wroclaw, Poland 2011
Abstract Islet transplantation has the potential to cure
type 1 diabetes. In recent years, the proportion of patients
achieving initial insulin independence has improved, but
longer term outcomes remain poor compared to those for
whole pancreas transplants. This review article will dis-
cuss factors affecting islet yield and viability leading up
to transplantation and in the immediate post-transplant
period.
Keywords Islet transplantation Apoptosis Hypoxia
Type 1 diabetes b cell
Proceedings of the Conference ‘‘Lymphocyte Development,
Tolerance and Autoimmunity: Solved and Open Questions’’,
Held at the Ludwik Hirszfeld Institute of Immunology
and Experimental Therapy, Polish Academy of Sciences,
on May 12, 2011 (Wrocław, Poland)
Lymphocyte Development, Tolerance and Autoimmunity: a personal perspective
Pawel Kisielow
Abstracts
Apoptosis in Lymphocytes of Pancreatic Cancer Patients:
Influence of Preoperative Enteral Immunonutrition
and Extensive Surgery
Robert Słotwin´ ski • Waldemar Olszewski • Maciej Słodkowski • Gustaw Lech •
Marzanna Zaleska • Sylwia Ke˛dziora • Anna Włuka • Anna Domaszewska •
Sylwia Słotwin´ ska • Wojciech Krasnode˛bski • Zdzisław Wo´jcik
Received: 6 August 2010 / Accepted: 8 April 2011 / Published online: 5 August 2011
Ó L. Hirszfeld Institute of Immunology and Experimental Therapy, Wroclaw, Poland 2011
Abstract The mechanisms of correcting immune disor-
ders in patients with pancreatic cancer requiring major
surgery procedures by introducing perioperative immune-
enhancing diet (immunonutrition) are still unclear. The
purpose of our study was to investigate the effect of pan-
creatic cancer, extensive surgery and immunonutrition
versus enteral standard nutrition on the apoptotic signaling
pathways. The randomized studies were performed in 72
patients before and after pancreatic cancer resection with
preoperative standard (Group I) or enteral immunonutrition
(Group II). The expressions of Bcl-2, Bax, caspases-3, -9,
NF-jB, PARP-1/89 kDa, TNFR1/CD120a and Fas/CD95
in peripheral blood lymphocytes were assessed by western
blot analysis and flow cytometry before and on day 1, 3 and
7 after surgery. In malnourished patients before and after
surgery, the expression of Bcl-2, Bax, NF-jB, PARP-1 was
significantly lower, whereas the expression of caspases, as
well as the percentage of cells with death receptors were
significantly higher when compared with the control group.
There was no difference in Bcl-2, Bax and PARP-1
expression between the control group and the patients with
normal nutritional status (Group III) before surgery. In
comparison to the standard nutrition, the preoperative im-
munonutrition increased the Bcl-2 and Bax expression
inconsiderably but significantly increased the percentage of
CD95- and CD120a-positive lymphocytes after surgery. In
malnourished patients with pancreatic cancer, the over-
whelming expression of caspases and the decrease
expression of anti-apoptotic proteins may lead to inappro-
priate lymphocyte apoptosis and higher cell depletion. The
preoperative enteral immunonutrition prevents the postop-
erative decrease in lymphocyte subsets, but a higher level
of lymphocyte susceptibility to undergo accelerated apop-
tosis can also be considered.
Keywords Pancreatic cancer Immunonutrition
Apoptosis
Bone Marrow Mesenchymal Cells: How Do They Contribute
to Tissue Repair and Are They Really Stem Cells?
Yasumasa Kuroda • Masaaki Kitada •
Shohei Wakao • Mari Dezawa
Received: 14 January 2011 / Accepted: 7 March 2011 / Published online: 26 July 2011
Ó L. Hirszfeld Institute of Immunology and Experimental Therapy, Wroclaw, Poland 2011
Abstract Adult stem cells typically generate the cell
types of the tissue in which they reside, and thus the range
of their differentiation is considered limited. Bone marrow
mesenchymal stem cells (MSCs) are different from other
somatic stem cells in that they differentiate not only into
the same mesodermal-lineage such as bone, cartilage, and
adipocytes but also into other lineages of ectodermal and
endodermal cells. Thus, MSCs are a unique type of adult
stem cells. In addition, MSCs home to damaged sites,
differentiate into cells specific to the tissue and contribute
to tissue repair. Therefore, application of MSCs in the
treatment of various diseases, including liver dysfunction,
myocardial infarction, and central nervous system repair,
has been initiated. Because MSCs are generally harvested
as adherent cells from bone marrow aspirates, however,
they comprise heterogeneous cell populations and their
wide-ranging differentiation ability and repair functions are
not yet clear. Recent evidence suggests that a very small
subpopulation of cells that assume a repair function with
the ability to differentiate into trilineage cells resides
among human MSCs and effective utilization of such cells
is expected to improve the repair effect of MSCs. This
review summarizes recent advances in the clarification of
MSC properties and discusses future perspectives.
Keywords Mesenchymal stem cells Adult stem cells
Transdifferenitation Cytokines Repair
Clinical Significance of the HLA-E and CD94/NKG2 Interaction
Milena Iwaszko • Katarzyna Bogunia-Kubik
Received: 4 January 2011 / Accepted: 26 April 2011 / Published online: 29 July 2011
Ó L. Hirszfeld Institute of Immunology and Experimental Therapy, Wroclaw, Poland 2011
Abstract HLA-E belongs to the non-classical HLA (class
Ib family) broadly defined by a limited polymorphism and
a restricted pattern of cellular expression. So far, only two
functional alleles differing at only one amino acid position
(non-synonymous mutation) in the a2 heavy chain domain,
where an arginine in position 107 in HLA-E*0101 is
replaced by a glycine in HLA-E*0103, have been reported.
The interaction between non-classical HLA-E molecule
and CD94/NKG2A receptor plays a crucial role in the
immunological response involving natural killer (NK) cells
and cytotoxic T lymphocytes. All proteins forming CD94/
NKG2 receptors are encoded by genes situated in the same
cluster on chromosome 12, allowing tight control over the
order of their expression. The inhibitory members of the
NKG2 receptor family are available on the cell surface
before activating the members to prevent autoimmune
incidents during immune cells’ ontogenesis. In the present
review, the potential role of this interaction in viral infec-
tion, pregnancy and transplantation of allogeneic
hematopoietic stem cells (HSC) is presented and discussed.
The review will also include the effect of HLA-E poly-
morphism on the outcome of HSC transplants in humans.
Keywords HLA-E CD94/NKG2 receptor
Pregnancy Viral infections Transplantation
of hematopoietic stem cells
Nuclear Factor-jB: a Key Regulator in Health and Disease
of Lungs
Sanjay Batra • Gayathriy Balamayooran •
Malaya K. Sahoo
Received: 7 January 2011 / Accepted: 2 March 2011 / Published online: 26 July 2011
Ó L. Hirszfeld Institute of Immunology and Experimental Therapy, Wroclaw, Poland 2011
Abstract Rel/NF-jB transcription factors play a key role
in modulating the response of immunoregulatory genes
including cytokines and chemokines, cell adhesion mole-
cules, acute phase proteins, and anti-microbial peptides.
Furthermore, an array of genes important for angiogenesis,
tumor invasion and metastasis is also regulated by nuclear
factor-jB (NF-jB). Close association of NF-jB with
inflammation and tumorigenesis makes it an attractive
target for basic research as well as for pharmaceutical
industries. Studies involving various animal and cellular
models have revealed the importance of NF-jB in patho-
biology of lung diseases. This review (a) describes
structures, activities, and regulation of NF-jB family
members; (b) provides information which implicates
NF-jB in pathogenesis of pulmonary inflammation and cancer;
and (c) discusses information about available synthetic and
natural compounds which target NF-jB or specific com-
ponents of NF-jB signal transduction pathway and which
may provide the foundation for development of effective
therapy for lung inflammation and bronchogenic
carcinomas.
Keywords NF-jB Inflammation Cancer Lung
Transitional B Cells: How Well Are the Checkpoints
for Specificity Understood?
Anna Vossenka¨mper • Jo Spencer
Received: 18 February 2011 / Accepted: 26 April 2011 / Published online: 26 July 2011
Ó L. Hirszfeld Institute of Immunology and Experimental Therapy, Wroclaw, Poland 2011
Abstract It is crucial for the immune system to minimise
the number of circulating mature self-reactive B cells, in
order to reduce the potential for the development of auto-
antibody-related autoimmune diseases. Studies of animal
models have identified two major checkpoints that ensure
that such cells do not contribute to the naı¨ve B cell rep-
ertoire. The first is in the bone marrow as B cells develop
and the second is in the spleen; B cells that are released
from the bone marrow as transitional B cells go through
more stringent selection in the spleen before they develop
into mature naı¨ve B cells. Transitional B cells and their
maturation have mostly been studied in mice. However,
recent studies characterised human transitional B cells and
found considerable differences to current models. In this
review, we will consider these differences alongside known
differences in mouse and human splenic function and ask
whether human transitional B cells might develop along a
different pathway.
Keywords Transitional B cells Spleen Checkpoint
Marginal zone
Expression of Th1/Th2/Th3/Th17-Related Genes
in Recurrent Aphthous Ulcers
Natalia Lewkowicz • Barbara Kur •
Anna Kurnatowska • Henryk Tchorzewski •
Przemyslaw Lewkowicz
Received: 4 January 2011 / Accepted: 28 April 2011 / Published online: 30 July 2011
Ó L. Hirszfeld Institute of Immunology and Experimental Therapy, Wroclaw, Poland 2011
Abstract The pathogenesis of recurrent aphthous ulcer-
ation (RAU) is unknown, although an abnormal immune
reaction appears to be involved. RAU may result from oral
epithelium damage caused by T cell-mediated immune
response. To improve understanding of the role of T cells
in RAU, the present study analyzed. the expression of T
cell-related genes in oral ulcers from patients with RAU, as
well as in healthy non-keratinized oral mucosa from
aphthae-free volunteers. Biopsies from RAU patients and
healthy individuals were analyzed using Human Th1-Th2-
Th3 RT2 Profiler PCR Array and qRT-PCR that allowed to
quantify the transcript levels of 86 genes related to T cell
activation. We found that cells present in aphthous ulcers
express a characteristic Th1-like gene profile. The majority
of genes up-regulated in aphthous lesions such as IFN-c,
TNF, IL-15, IRF1, STAT-1 and STAT-4 were Th1-asso-
ciated. Th2-realated genes were not overexpressed in RAU
tissues, with the exception for CCR3. Th3- and Th17-
related gene expression patterns were not demonstrated in
RAU. These findings clearly reveal that aphthous ulcer
formation is predominantly dependent on the activation of
the Th1-type immune response.
Keywords Recurrent aphthous ulcers Oral mucosa
Th-related gene expression
Soluble TNF-a Receptor I Encoded on Plasmid Vector
and Its Application in Experimental Gene Therapy
of Radiation-Induced Lung Fibrosis
Małgorzata Przybyszewska • Joanna Miłoszewska • Sylwia Rzon´ ca •
Halina Trembacz • Kazimiera Pys´niak • Agnieszka Kotlarz •
Paweł Swoboda • Marta Zalewska • Maciej Małecki
Received: 10 November 2010 / Accepted: 7 March 2011 / Published online: 3 June 2011
Ó L. Hirszfeld Institute of Immunology and Experimental Therapy, Wroclaw, Poland 2011
Abstract Post-radiation inflammatory reaction leads to
an irreversible pulmonary fibrosis which may cause lethal
respiratory insufficiency. Pathological inflammatory and
fibrotic changes might be attenuated by inhibiting tumour
necrosis factor (TNF)-a activity using TNF-a soluble
receptors. Thus, an experimental antifibrotic gene therapy
with the plasmid vector encoding a mouse soluble receptor
I for TNF-a (psTNFR-I) was assessed. Soluble TNFR-I
encoding gene was cloned into pcDNA3.1 plasmid. The
ability of psTNFR-I expressing vector to transfect cells,
and its biological activity in vitro and in vivo were
examined by PCR, RT-PCR, MTT assay and ELISA. The
C57Bl/6J mice received single intramuscular injection of
psTNFR-I, conjugated with polyetylenimine (PEI) 25 kDa,
equally divided to both hind legs, 3 days before irradiation
(20 Gy, Co60), and either a single injection or ten injec-
tions once a week after irradiation. The data proved the
effectiveness of psTNFR-I product to neutralise TNF-a
activity in vitro. The in vivo plasmid incorporation and
maintenance was confirmed. Measurements of plasma
soluble TNFR-I levels showed that the in vivo gene
transfer was effective. PEI was found to enhance trans-
fection efficiency in vivo. The psTNFR-I/PEI complexes
caused no toxicity in the transfected mice. C57Bl/6J mice
that received prolonged psTNFR-I/PEI injections devel-
oped lethal fibrotic syndrome and died 8 weeks later than
the mice treated with a double plasmid injection and the
control mice treated with a control plasmid. Sequential
administration of soluble TNFR-I by a nonviral, intra-
muscular gene transduction in the early and late post-
radiation inflammatory phase prolonged survival of
irradiated mice and attenuated the symptoms of lung
fibrosis. The psTNFR-I gene transduction may provide a
safe and simple method to partially neutralise TNF-a
activity and prevent radiation-induced lung injury.
Keywords Soluble TNFR-I psTNFR-I TNF-a
Post-radiation lung fibrosis Plasmid expressing vector
Gene therapy
Targeting NF-jB and HIF-1 Pathways for the Treatment
of Cancer: Part II
Jacek Wilczynski • Markus Duechler •
Malgorzata Czyz
Received: 18 November 2010 / Accepted: 2 March 2011 / Published online: 28 May 2011
Ó L. Hirszfeld Institute of Immunology and Experimental Therapy, Wroclaw, Poland 2011
Abstract Hypoxia that originates from disturbed growth
of solid tumors initiates a cascade of intracellular events
engaging hypoxia-inducible factors, HIF-1 and HIF-2.
Overexpression of HIF has been confirmed in solid tumors
and was unfortunately accompanied with chemo- and
radioresistance observed in many patients. Multiple cellu-
lar pathways resulting in HIF activation could be suc-
cessfully inhibited by use of different kinds of drugs (e.g.
topotecan, heat shock protein 90 and mTOR inhibitors,
YC-1, pleurotin or 2-methoxyestradiol), which are being
subjected into intensive investigation in clinical trials.
Keywords HIF inhibitors Cancer Treatment
Targeting NF-jB and HIF-1 Pathways for the Treatment
of Cancer: Part I
Jacek Wilczynski • Markus Duechler •
Malgorzata Czyz
Received: 18 November 2010 / Accepted: 2 March 2011 / Published online: 28 May 2011
Ó L. Hirszfeld Institute of Immunology and Experimental Therapy, Wroclaw, Poland 2011
Abstract The process of chronic inflammation is a common
link which connects different kinds of environmental pollu-
tants and infections with tumorigenesis. Transcription factor
NF-jB is a common final target for many inflammatory and
cell proliferation pathways, independent of the source of
stimuli (e.g., cytokines, growth factors, environmental car-
cinogens, radiation, hypoxia, bacteria, and viruses). Over-
activation of NF-jB has been confirmed in many tumors,
resulting in worse prognosis for patient survival. Therefore,
inhibition of cellular pathways for NF-jB activation is now-
adays considered as a promising anti-cancer therapy and is
extensively studied in clinical trials, or even has been adopted
as an approved therapy in some kinds of cancer.
Keywords NF-jB inhibitors Cancer Treatment
Recent Progress of Shiga Toxin Neutralizer for Treatment
of Infections by Shiga Toxin-Producing Escherichia coli
Kiyotaka Nishikawa
Received: 18 November 2010 / Accepted: 15 December 2010 / Published online: 5 June 2011
Ó L. Hirszfeld Institute of Immunology and Experimental Therapy, Wroclaw, Poland 2011
Abstract Infection with Shiga toxin (Stx)-producing
Escherichia coli (STEC), including O157:H7, causes
bloody diarrhea and hemorrhagic colitis in humans, occa-
sionally resulting in fatal systemic complications, such as
neurological damage and hemolytic-uremic syndrome.
Because Stx is a major virulence factor of the infectious
disease, a series of Shiga toxin neutralizers with various
structural characteristics has been developed as promising
therapeutic agents. Most of these agents function to bind to
the toxin directly and inhibit the binding to its receptor
present on the target cells. Other neutralizers do not inhibit
receptor binding but induce aberrant intracellular transport
of the toxin, resulting in effective detoxification. Such a
novel type of Stx neutralizer provides a new therapeutic
strategy against STEC infections. Here, recent progress of
the development of Stx neutralizers is reviewed.
Keywords Shiga toxin Neutralizer
Shiga toxin-producing E. coli Globotriaosyl ceramide
Peptide-library Clustering effect
Cancer Vaccines. Any Future?
Lukasz A. Myc • Andrzej Gamian •
Andrzej Myc
Received: 3 November 2010 / Accepted: 2 February 2011 / Published online: 5 June 2011
Ó L. Hirszfeld Institute of Immunology and Experimental Therapy, Wroclaw, Poland 2011
Abstract The idea that vaccination can be used to
fight cancer is not new. Approximately 100 years ago,
researchers attempted to stimulate a tumor-specific, thera-
peutic immune response to tumors by injecting patients
with cells and extracts from their own tumors, or tumors of
the same type from different individuals. During the last
decade, great efforts have been made to develop immu-
notherapeutic approaches for the treatment of malignant
diseases as alternatives to traditional chemo- and radio-
therapy. A quintessential goal of immunotherapy in cancer
is treatment with vaccines that elicit potent anti-tumor
immune responses without side effects. In this article, we
have attempted to review some of the most problematic
issues facing the development of cancer vaccines. With the
prospect of immunosuppression, an ill-designed cancer
vaccine can be more harmful than a no-benefit therapy. We
have noted that ‘‘immunoediting’’ and ‘‘immunodomi-
nance’’ are the premier setbacks in peptide-based vaccines
and therefore it appears necessary not only to manipulate
the activity of a vast number of principal components but
also to finely tune their concentrations in time and space. In
the face of all these quandaries, it is at least doubtful that
any reliable anti-cancer vaccine strategy will emerge in the
near future.
Keywords Adjuvant Liposomes Nanoemulsions
Tumor-specific antigens Tumor-associated antigens
Heteroclitic peptides Immunoediting Cytokines
TH1 TH2
Prevention of Lyme Disease: Promising Research
or Sisyphean Task?
Michal Krupka • Katerina Zachova •
Evzen Weigl • Milan Raska
Received: 2 December 2010 / Accepted: 2 March 2011 / Published online: 3 June 2011
Ó L. Hirszfeld Institute of Immunology and Experimental Therapy, Wroclaw, Poland 2011
Abstract Borrelia burgdorferi sensu lato (Spirochaetes)
is a group of at least 12 closely related species, some of
which are responsible for chronic zoonotic infection that
may cause Lyme disease. The only experimentally con-
firmed vector transmitting Borrelia to mammals is the
Ixodes ticks. Borrelia is a highly adapted pathogen that can
survive in the host organism in spite of the intense immune
responses. Some patients have chronic long-lasting com-
plications despite antibiotic therapy, probably due to
adverse effects of the immune responses. A preventive
vaccine against this bacterium has not been available due
to the relatively broad spectrum and antigenic variability of
Borrelia-surface lipoproteins and the different epitope
recognition by experimental animals and humans.
Although a human vaccine was marketed in the USA, it has
been already pulled off the market. In addition, this vaccine
was effective only in the USA, where the only pathogenic
species is B. burgdorferi sensu stricto. Recent data indicate
that a broadly effective vaccine will to be composed of a
mixture of several antigens or multiple epitopes.
Keywords Borrelia burgdorferi Lyme disease
Vaccine Osp
Increased Frequency of CD4+CD25highFoxP3+ Regulatory
T Cells in Patients with Hepatocellular Carcinoma
Xi Feng • Bo Li • Hui Ye • Dan Long
Received: 3 November 2010 / Accepted: 28 February 2011 / Published online: 3 June 2011
Ó L. Hirszfeld Institute of Immunology and Experimental Therapy, Wroclaw, Poland 2011
Abstract Accumulating evidence suggests regulatory T
cells (Tregs) are associated with impaired antitumor
responses. However, the relationship between the
CD4?CD25highFoxP3? Treg and hepatocellular carcinoma
(HCC) has not been well investigated. Levels of
CD4?CD25highFoxP3? Tregs in peripheral blood mono-
nuclear cells (PBMCs) from HCC patients and healthy
donors, tumor infiltrating lymphocytes (TILs) extracted
from HCC, and hepatic lymphocytes extracted from
resected liver were measured by flow cytometry, and their
effects on T-cell proliferation was determined by 3H-thy-
midine incorporation. Serum levels of interleukin (IL)-10
and transforming growth factor (TGF)-b1 were measured
by enzyme linked immunosorbent assay. The frequency of
Tregs in PBMCs from HCC patients was higher than that
from healthy donors. Similarly, the frequency of Tregs in
TILs was higher than that of hepatic lymphocytes. On the
other hand, the 3H-thymidine uptake by TILs and PBMCs
from HCC patients was decreased drastically when
compared to the counterparts from normal controls. Fur-
thermore, serum IL-10 and TGF-b1 levels increased
significantly in HCC patients when compared to the heal-
thy donors. This study identified an increased frequency of
CD4?CD25highFoxP3? Tregs in patients with HCC. The
elevated serum IL-10, TGF-b1 levels also correlated with
impaired antitumor responses in these patients. Further
effort is needed to establish new immunotherapeutic
strategies designed to modulate Tregs to promote a com-
petent antitumor response.
Keywords CD4?CD25highFoxP3? Treg
Hepatocellular carcinoma IL-10 TGF-b1
Bim: Guardian of Tissue Homeostasis and Critical Regulator
of the Immune System, Tumorigenesis and Bone Biology
Toru Akiyama • Sakae Tanaka
Received: 28 September 2010 / Accepted: 11 January 2011 / Published online: 3 June 2011
Ó L. Hirszfeld Institute of Immunology and Experimental Therapy, Wroclaw, Poland 2011
Abstract One of the most important roles of apoptosis is
the maintenance of tissue homeostasis. Impairment of
apoptosis leads to a number of pathological conditions. In
response to apoptotic signals, various proteins are activated
in a pathway and signal-specific manner. Recently, the pro-
apoptotic molecule Bim has attracted increasing attention
as a pivotal regulator of tissue homeostasis. The Bim
expression level is strictly controlled in both transcriptional
and post-transcriptional levels. This control is dependent
on cell, tissue and apoptotic stimuli. The phenotype of
Bim-deficient mice is a systemic lupus erythematosus-like
autoimmune disease with an abnormal accumulation of
hematopoietic cells. Bim is thus a critical regulator of
hematopoietic cells and immune system. Further studies
have revealed the critical roles of Bim in various normal
and pathological conditions, including bone homeostasis
and tumorigenesis. The current understanding of Bim sig-
naling and roles in the maintenance of tissue homeostasis is
reviewed in this paper, focusing on the immune system,
bone biology and tumorigenesis to illustrate the diversified
role of Bim.
Keywords Bim Apoptosis Tissue homeostasis
Immune system Tumorigenesis Bone biology
The Patterns of MHC Association in Aplastic and Non-aplastic
Paroxysmal Nocturnal Hemoglobinuria
Jacek Nowak • Renata Mika-Witkowska • Ewa Mendek-Czajkowska • Marta Rogatko-Koros´ •
El_zbieta Graczyk-Pol • Hanna Pyl • Aneta Klimczak • Małgorzata Wo´jcik •
Monika Prochorec-Sobieszek • Renata Maryniak • Barbara _Zupan´ ska
Received: 26 May 2010 / Accepted: 27 December 2010 / Published online: 27 March 2011
Ó L. Hirszfeld Institute of Immunology and Experimental Therapy, Wroclaw, Poland 2011
Abstract The deficiency of glycosyl-phosphatidylinosi-
tol (GPI)-anchored proteins in plasma membranes of
PIG–A gene mutated hematopoietic stem cells (HSCs) is so
far insufficient to explain the domination of paroxysmal
nocturnal hemoglobinuria (PNH) clone over the normal
HSC. We attempted to elucidate possible link between
MHC and initial severe aplastic anemia (ISAA/PNH) type
and non-aplastic (n/PNH) outcome of PNH. In 50 PNH
patients assigned as ISAA/PNH (n = 13), n/PNH (n = 33)
or nonassigned (n = 4) and 200 ethnically matched con-
trols we analyzed MHC associations. Our data confirmed
strong associations of DRB1*15:01 (RR = 3.51,
p = 0.0011) and DQB1*06:02 (RR = 7.09, p = 0.000026)
alleles, especially with n/PNH subtype. B*18:01 allele was
associated with increased risk of ISAA/PNH subtype
(RR = 5.25, p = 0.0028). We conclude that both class II
and class I MHC alleles are associated with different
subsets of PNH. Clonal selection of PIG–A mutated cells
with cognate metabolic block is associated with MHC class
II alleles DRB1*15:01 and DQB1*06:02 independent from
initial severe AA clone selection. MHC class I molecule
B*18:01 can additionally influence the domination of PNH
clone in PNH subjects with initial severe aplastic anemia.
Keywords Paroxysmal nocturnal hemoglobinuria
Major histocompatibility complex Association study
Bone marrow aplasia
Gender Specific Differences in the Immune Response to Infection
Erin E. McClelland • Jennifer M. Smith
Received: 29 September 2010 / Accepted: 15 December 2010 / Published online: 26 March 2011
Ó L. Hirszfeld Institute of Immunology and Experimental Therapy, Wroclaw, Poland 2011
Abstract There are many instances where males and
females differ in the susceptibility to infections. The reason
for these differences in susceptibility is multifactorial. The
primary cause is thought to be due to differences induced
by sex hormones and their effects on gene expression as
well as the immune system, but may also be due to innate
physiological differences between males and females. This
review summarizes gender specific differences seen in
infections caused by bacteria, fungi, parasites and viruses.
Ultimately, gender specific differences appear to be
dependent on the microbe causing the infection, as not
every infection with a specific microbial type results in
increased susceptibility of one gender over the other. This
suggests that there is an interaction between gender specific
immune differences and the specific immune response to
individual microbes.
Keywords Gender differences Microbial infection
Immune response
Phosphatidylserine-Containing Liposomes: Potential
Pharmacological Interventions Against Inflammatory
and Immune Diseases Through the Production of Prostaglandin
E2 After Uptake by Myeloid Derived Phagocytes
Zhou Wu • Hiroshi Nakanishi
Received: 28 September 2010 / Accepted: 5 November 2010 / Published online: 10 April 2011
Ó L. Hirszfeld Institute of Immunology and Experimental Therapy, Wroclaw, Poland 2011
Abstract Phosphatidylserine (PS), which is normally
located on the inner leaflet of the plasma membrane,
translocates to the outer leaflet at the early stage of apop-
tosis. The PS externalization provides a signal for
phagocytes to initiate uptake of apoptotic cells. After
phagocytosis of apoptotic cells, phagocytes induce the
secretion of anti-inflammatory mediators including pros-
taglandin E2 (PGE2). PS-containing liposomes (PSLs) can
mimic the effects of apoptotic cells on phagocytes to
induce the secretion of PGE2. PSLs induce the PGE2
secretion from microglia without induction of either
cyclooxygenase (COX)-2 or microsomal prostaglandin E
synthase (mPGES)-1. PSLs are found to rather utilize
COX-1/mPGES-2 system to produce PGE2 secretion and
then shift microglia and macrophages from pro- to anti-
inflammatory phenotype by an autocrine action of PGE2.
Moreover, PSLs inhibit the maturation of dendritic cells
and osteoclast precursors. Therefore, PSLs will be potential
pharmacological interventions for inflammatory and
immune diseases through feedback mechanism utilizing
PGE2.
Keywords Phosphatidylserine (PS)
PS-containing liposomes Phagocytes PGE2
Gut Microbiota, Probiotics and Inflammatory Bowel Disease
Johannes Stephani • Katarina Radulovic •
Jan Hendrik Niess
Received: 19 July 2010 / Accepted: 13 December 2010 / Published online: 29 March 2011
Ó L. Hirszfeld Institute of Immunology and Experimental Therapy, Wroclaw, Poland 2011
Abstract The colonization of humans with commensals
is critical for our well-being. This tightly regulated sym-
biotic relationship depends on the flora and an intact
mucosal immune system. A disturbance of either com-
pound can cause intestinal inflammation. This review
summarizes extrinsic and intrinsic factors contributing to
intestinal dysbiosis and inflammatory bowel disease.
Keywords Commensals Gut immunology Probiotics
Dendritic cells Macrophages
Ethical Considerations in Face Transplantation: Ethical Issues
Related to Inclusion Criteria for Face Transplant Candidates
Maria Siemionow
Myelin Restoration: Progress and Prospects for Human Cell
Replacement Therapies
Gregory B. Potter • David H. Rowitch •
Magdalena A. Petryniak
Received: 15 July 2010 / Accepted: 17 November 2010 / Published online: 2 April 2011
Ó L. Hirszfeld Institute of Immunology and Experimental Therapy, Wroclaw, Poland 2011
Abstract Oligodendrocytes are the primary source of
myelin in the adult central nervous system (CNS), and their
dysfunction or loss underlies several diseases of both
children and adults. Dysmyelinating and demyelinating
diseases are thus attractive targets for cell-based strategies
since replacement of a single presumably homogeneous
cell type has the potential to restore functional levels of
myelin. To understand the obstacles that cell-replacement
therapy might face, we review oligodendrocyte biology and
emphasize aspects of oligodendrocyte development that
will need to be recapitulated by exogenously transplanted
cells, including migration from the site of transplantation,
axon recognition, terminal differentiation, axon wrapping,
and myelin production and maintenance. We summarize
studies in which different types of myelin-forming cells
have been transplanted into the CNS and highlight the
continuing challenges regarding the use of cell-based
therapies for human white matter disorders.
Keywords Myelin Transplantation Oligodendrocyte
Development Pelizaeus–Merzbacher disease
Cell-replacement Leukodystrophy
Glucose Metabolism Disorders and the Risk of Cancer
Paweł Pia˛tkiewicz • Anna Czech
Received: 15 September 2010 / Accepted: 29 November 2010 / Published online: 30 March 2011
Ó L. Hirszfeld Institute of Immunology and Experimental Therapy, Wroclaw, Poland 2011
Abstract Diabetes and cancer are diseases which take the
size of an epidemic spread across the globe. Those diseases
are influenced by many factors, both genetic and environ-
mental. Precise knowledge of the complex relationships
and interactions between these two conditions is of great
importance for their prevention and treatment. Many epi-
demiological studies have shown that certain types of
cancer, especially gastrointestinal cancers (pancreas, liver,
colon) and also the urinary and reproductive system can-
cers in women are more common in patients with diabetes
or related metabolic disorders. There are also studies
showing the inverse relationship between diabetes and
cancer, or the lack of it, but they are less numerous and
relate mainly to prostate cancer or squamous cell carci-
noma of the esophagus. Epidemiological studies, however,
do not say anything about the mechanisms of these
dependencies. For this purpose, molecular research is
needed on the metabolism of cells (including tumor cells)
and on metabolic dysfunctions that arise due to changes in
the cell environment taking place in the sick, as well as in
the intensely treated human organism.
Keywords Glucose Insulin Diabetes Cancer
cd T cell Receptor Ligands and Modes of Antigen Recognition
Eric Champagne
Received: 30 August 2010 / Accepted: 2 December 2010 / Published online: 6 February 2011
Ó L. Hirszfeld Institute of Immunology and Experimental Therapy, Wroclaw, Poland 2011
Abstract T lymphocytes expressing the cd-type of T cell
receptors (TCRs) for antigens contribute to all aspects of
immune responses, including defenses against viruses,
bacteria, parasites and tumors, allergy and autoimmunity.
Multiple subsets have been individualized in humans as
well as in mice and they appear to recognize in a TCR-
dependent manner antigens as diverse as small non-peptidic
molecules, soluble or membrane-anchored polypeptides
and molecules related to MHC antigens on cell surfaces,
implying diverse modes of antigen recognition. We review
here the cd TCR ligands which have been identified along
the years and their characteristics, with emphasis on a few
systems which have been extensively studied such as
human cd T cells responding to phosphoantigens or murine
cd T cells activated by allogeneic MHC antigens. We dis-
cuss a speculative model of antigen recognition involving
simultaneous TCR recognition of MHC-like and non-MHC
ligands which could fit with most available data and shares
many similarities with the classical model of MHC-
restricted antigen recognition for peptides or lipids by T
cells subsets with ab-type TCRs.
Keywords Gamma delta lymphocytes T cell receptor
Innate immunity Antigen recognition
Major histocompatibility complex
Innate Immunity Signaling Pathways: Links between
Immunonutrition and Responses to Sepsis
Robert Słotwin´ ski • Sylwia Słotwin´ ska •
Sylwia Ke˛dziora • Barbara-Joanna Bałan
Received: 20 March 2010 / Accepted: 30 July 2010 / Published online: 6 February 2011
Ó L. Hirszfeld Institute of Immunology and Experimental Therapy, Wroclaw, Poland 2011
Abstract Septic infections in patients treated in intensive
care units show the highest mortality rates. Despite
advances in treatment methods, there is still no therapy
available to efficiently reduce the excessive inflammatory
response, which can increase the risk of multiple organ
failure. One of the ways to discover new, more efficient
treatment methods involves regulating the mechanisms of
inflammatory response to a massive infection. Toll-like
receptors (TLRs) that recognize pathogen-associated
molecular patterns play a significant role in innate anti-
bacterial and inflammatory responses. The regulatory
impact of immunonutrition on TLR expression in septic
patients seems to be a promising research direction. This
paper presents the main mechanisms for the innate immune
response to lipopolysaccharide, based on the research
results for both TLR-dependent and independent signaling
pathways. Special emphasis was put on the research results
for the TLR-dependent immune response and the anti-
bacterial/anti-inflammatory response after applying immuno-
nutrition with increased concentrations of glutamine and
unsaturated fatty acids.
Keywords Innate immunity Immunonutrition
Toll-like receptors Sepsis
Pathogenesis of Joint Destruction in Rheumatoid Arthritis
Shunichi Shiozawa • Ken Tsumiyama •
Kohsuke Yoshida • Akira Hashiramoto
Received: 23 June 2010 / Accepted: 20 September 2010 / Published online: 16 February 2011
Ó L. Hirszfeld Institute of Immunology and Experimental Therapy, Wroclaw, Poland 2011
Abstract Synovial mesenchymal cells, matrix metallopro-
teinases (MMPs), and osteoclasts are the three major players
directly responsible for the pathogenesis of rheumatoid joint
destruction. First, synovial mesenchymal cells, internally
driven by a transcription factor c-Fos/AP-1, not only directly
invade cartilage and bone as a granulation tissue called
‘‘pannus’’ but also release inflammatory cytokine interleukin
(IL)-1b. IL-1b induces MMPs and activates osteoclasts.
Synovial cells can also present antigen to T cells to drive
antigen-specific immune responses. Second, cartilaginous
joint matrix can only be degraded after the first attack of
collagen fibrils by MMPs, and importantly, most of the MMPs
are under the control of c-Fos/AP-1 and IL-1b as well. Third,
differentiation of osteoclast is driven internally by NFATc1,
where NFATc1 is under the control of TRAF6, c-Fos/AP-1
and osteoclastogenic signaling complex. IL-1b has been
shown to induce osteoclastogenesis directly and also indi-
rectly via signaling through RANKL. Therefore, IL-1b and
c-Fos/AP-1 influence each other’s gene expression and
activity, resulting in an orchestrated cross-talk that is crucial to
arthritic joint destruction, and thus, blockade of IL-1b and/or
c-Fos/AP-1 can be most promising as a therapeutic target, and
in fact, a selective inhibition of c-Fos/AP-1 does resolve
arthritic joint destruction.
Keywords Rheumatoid arthritis Joint destruction
c-Fos c-Fos/AP-1 IL-1beta
CXCL8 and CCL5 Expression in Synovial Fluid and Blood Serum
in Patients with Osteoarthritis of the Knee
Adam W. Pierzchala • Damian J. Kusz •
Grzegorz Hajduk
Received: 13 May 2010 / Accepted: 2 December 2010 / Published online: 19 February 2011
Ó L. Hirszfeld Institute of Immunology and Experimental Therapy, Wroclaw, Poland 2011
Abstract Chemokines proved able to induce release of
enzymes relevant in cartilage damage. The present study
addressed the levels of CXCL8 and CCL5 and the potential
role of these chemokines in predicting the morphological
changes in the course of osteoarthritis (OA). Synovial fluid
(SF) and blood serum were obtained from 20 patients
undergoing knee replacement surgery because of OA. For
comparison, samples were also obtained from another 20
patients during diagnostic or therapeutic arthroscopy per-
formed because of knee injury. The samples were analyzed
for CXCL8 and CCL5 using enzyme-linked immunosorbent
assay. SF from the group with OA showed significantly
(p = 0.024) increased levels of CXCL8 when compared
with the group after knee injury. We have not demonstrated
any significant correlation between chemokine expression
and clinical or radiological signs of OA. Mediators of
inflammation are the potential predicting factors of OA,
however, with respect to examined chemokines develop-
ment of a diagnostic test can be limited by the low serum
concentration and lack of correlation with clinical and
radiological signs of the disease.
Keywords Osteoarthritis Cytokines Chemokines
Knee injuries
Physiological Roles of Mammalian Sulfate Transporters
NaS1 and Sat1
Daniel Markovich
Received: 12 August 2010 / Accepted: 8 October 2010 / Published online: 6 February 2011
Ó L. Hirszfeld Institute of Immunology and Experimental Therapy, Wroclaw, Poland 2011
Abstract This review summarizes the physiological roles
of the renal sulfate transporters NaS1 (Slc13a1) and Sat1
(Slc26a1). NaS1 and Sat1 encode renal anion transporters
that mediate proximal tubular sulfate reabsorption and
thereby regulate blood sulfate levels. Targeted disruption
of murine NaS1 and Sat1 leads to hyposulfatemia and
hypersulfaturia. Sat1 null mice also exhibit hyperoxalemia,
hyperoxaluria and calcium oxalate urolithiasis. Dysregu-
lation of NaS1 and Sat1 leads to hypersulfaturia,
hyposulfatemia and liver damage. Loss of Sat1 leads
additionally to hyperoxaluria with hyperoxalemia, neph-
rocalcinosis and calcium oxalate urolithiasis. These data
indicate that the renal anion transporters NaS1 and Sat1 are
essential for sulfate and oxalate homeostasis, respectively.
Keywords Sulfate transport Oxalate transport
Renal reabsorption Anion exchange NaS1 Sat1
SLC13A1 SLC26A1
The Cystic Fibrosis Neutrophil: A Specialized Yet Potentially
Defective Cell
Elaine Hayes • Kerstin Pohl • Noel G. McElvaney •
Emer P. Reeves
Received: 1 July 2010 / Accepted: 27 October 2010 / Published online: 11 February 2011
Ó L. Hirszfeld Institute of Immunology and Experimental Therapy, Wroclaw, Poland 2011
Abstract Cystic fibrosis (CF) is one of the commonest
genetically inherited diseases in the world. It is character-
ized by recurrent respiratory tract infections eventually
leading to respiratory failure. One of the hallmarks of this
disease is a persistent and predominantly neutrophil driven
inflammation. Neutrophils provide the first line of defence
by killing and digesting phagocytosed bacteria and fungi,
yet despite advances in our understanding of the molecular
and cellular basis of CF, there remains a paradox of why
recruited CF neutrophils fail to eradicate bacterial infec-
tions in the lung. This review describes mechanisms
involved in neutrophil migration, microbial killing and
apoptosis leading to inflammatory resolution. We discuss
dysregulated neutrophil activity and consider genetic ver-
sus inflammatory neutrophil reprogramming in CF and
ultimately pharmacological modulation of the CF neutro-
phil for therapeutic intervention.
Keywords Cystic fibrosis Neutrophils
NADPH oxidase Neutrophil elastase
Protein Kinase C Isoforms in Neutrophil Adhesion and Activation
Anna Bertram • Klaus Ley
Received: 14 June 2010 / Accepted: 8 October 2010 / Published online: 6 February 2011
Ó L. Hirszfeld Institute of Immunology and Experimental Therapy, Wroclaw, Poland 2011
Abstract Neutrophils are the first line of defense against
bacterial and mycotic pathogens. In order to reach the
pathogens, neutrophils need to transmigrate through the
vascular endothelium and migrate to the site of infection.
Defense strategies against pathogens include phagocytosis,
production and release of oxygen radicals through the
oxidative burst, and degranulation of antimicrobial and
inflammatory molecules. Protein kinase C (PKC)-d is
required for full assembly of NADPH oxidase and activa-
tion of the respiratory burst. Neutrophils also express PKC-
a and -b, which may be involved in adhesion, degranula-
tion and phagocytosis, but the evidence is not conclusive
yet. This review focuses on the potential impact of protein
kinase C isoforms on neutrophil adhesion and activation.
Keywords Neutrophil Protein kinase C
Integrin activation Adhesion Rolling NADPH oxidase