Pretreatment Levels of Vascular Endothelial Growth Factor in Plasma Predict a Complete Remission Rate and Time to Relapse or Progression in Patients with Diffuse Large B-Cell Lymphoma
Ewa Lech-Maranda • Jacques Bienvenu • Florence Broussais-Guillaumot • Anne-Sophie Michallet • Krzysztof Warzocha • Przemysław Biliński •Peter Boyle • Bertrand Coiffier • Gilles Salles
Abstract The aim of this study was to verify whether pretreatment plasma levels of vascular endothelial growth factor (VEGF) correlate with prognosis and survival of patients with diffuse large B-cell lymphoma (DLBCL). Plasma VEGF levels were assessed at the time of diagnosis in 157 DLBCL patients treated with anthracycline-based chemotherapy. Plasma VEGF levels greater than or equal to the highest quartile (high VEGF levels) were associated with lower probability of a complete remission achievement (odds ratio 0.3; 95 % confidence interval [CI]: 0.1–0.6; p = 0.002) in univariate as well as in multivariate analysis (p = 0.04). The estimated 3-year progression-free survival (PFS) rate of patients with high VEGF levels was 31.7 % (95 % CI 17–51) compared to the 62.5 % 3-year PFS rate (95 % CI 53–71; p = 0.0004) in the patients with lower values. The former group of patients demonstrated an estimated 3-year overall survival (OS) rate of 47.1 % (95 % CI 30–65) in contrast to the 3-year OS rate of 64.3 % (95 % CI 54–73; p = 0.02) in the latter. In multivariate analysis, the high VEGF level retained its independent impact on shorter PFS (p = 0.02). Our results suggest that VEGF plays an important role in the clinical course of DLBCL. VEGF may be a useful marker for selecting the patients for whom new treatment approaches, especially those based on VEGF inhibitors, could be recommended.
Keywords Vascular endothelial growth factor Diffuse large B-cell lymphoma Chemotherapy Prognosis
Multi-Step Regulation of Interferon Induction by Hepatitis C Virus
Hiroyuki Oshiumi • Kenji Funami • Hussein H. Aly • Misako Matsumoto • Tsukasa Seya
Abstract Acute hepatitis C virus (HCV) infection evokes several distinct innate immune responses in host, but the virus usually propagates by circumventing these responses. Although a replication intermediate double-stranded RNA is produced in infected cells, type I interferon (IFN) induction and immediate cell death are largely blocked in infected cells. In vitro studies suggested that type I and III IFNs are mainly produced in HCV-infected hepatocytes if the MAVS pathway is functional, and dysfunction of this pathway may lead to cellular permissiveness to HCV replication and production. Cellular immunity, including natural killer cell activation and antigen-specific CD8 T-cell proliferation, occurs following innate immune activation in response to HCV, but is often ineffective for eradication of HCV. Constitutive dsRNA stimulation differs in output from type I IFN therapy, which has been an authentic therapy for patients with HCV. Host innate immune responses to HCV RNA/proteins may be associated with progressive hepatic fibrosis and carcinogenesis once persistent HCV infection is established in opposition to the IFN system. Hence, innate RNA sensing exerts pivotal functions against HCV genome replication and host pathogenesis through modulation of the IFN system. Molecules participating in the RIG-I and Toll-like receptor 3 pathways are the main targets for HCV, disabling the anti-viral functions of these IFN-inducing molecules. We discuss the mechanisms that abolish type I and type III IFN production in HCV-infected cells, which may contribute to understanding the mechanism of virus persistence and resistance to the IFN therapy.
Keywords Hepatitis C virus TLR3 TICAM-1 (TRIF) MAVS (IPS-1, Cardif, VISA) Interferon-inducing pathway Double-stranded RNA
Differentiation of Stem Cells into Insulin-Producing Cells:
Current Status and Challenges
Marta Pokrywczynska • Sandra Krzyzanowska •
Arkadiusz Jundzill • Jan Adamowicz •
Tomasz Drewa
Received: 1 June 2012 / Accepted: 20 December 2012 / Published online: 3 January 2013
Ó L. Hirszfeld Institute of Immunology and Experimental Therapy, Wroclaw, Poland 2012
Abstract Diabetes mellitus is one of the most serious
public health challenges of the twenty-first century. Allo-
genic islet transplantation is an efficient therapy for type 1
diabetes. However, immune rejection, side effects of
immunosuppressive treatment as well as lack of sufficient
donor organs limits its potential. In recent years, several
promising approaches for generation of new pancreatic b
cells have been developed. This review provides an over-
view of current status of pancreatic and extra-pancreatic
stem cells differentiation into insulin-producing cells and
the possible application of these cells for diabetes treat-
ment. The PubMed database was searched for English
language articles published between 2001 and 2012, using
the keyword combinations: diabetes mellitus, differentia-
tion, insulin-producing cells, stem cells.
Keywords Diabetes mellitus Differentiation
Insulin-producing cells Stem cells
Granulocyte-Colony Stimulating Factor as Treatment
Option in Patients with Recurrent Miscarriage
Claudia Santjohanser • Catherine Knieper •
Cordula Franz • Kaino Hirv • Osama Meri •
Manfred Schleyer • Wolfgang Wu¨ rfel • Bettina Toth
Received: 11 May 2012 / Accepted: 20 December 2012 / Published online: 24 January 2013
Ó L. Hirszfeld Institute of Immunology and Experimental Therapy, Wroclaw, Poland 2013
Abstract In 1–5 % of patients during childbearing years
recurrent miscarriages (RM) occur. There are established
risk factors like anatomical, endocrine and hemostatic
disorders as well as immunological changes in the maternal
immune system. Nevertheless, further elucidation of the
pathogenesis remains a matter of debate. In addition, there
are no standardized immunological treatment strategies.
Recent studies indicate possible effects of tumor necrosis
factor a blocker and granulocyte-colony stimulating factor
(G-CSF) concerning live birth rate (LBR) in RM patients.
Therefore, we performed a retrospective cohort study in
patients undergoing assisted reproductive treatment (ART)
with known RM analysing the possible benefits of G-CSF
application. From January 2002 to December 2010, 127
patients (199 cylces) with RM (at least 2 early miscar-
riages) 49 (72 cycles) receiving G-CSF and 78 (127 cycles)
controls receiving either no medication (subgroup 1) or
Cortisone, intravenous immunoglobulins or low molecular
weight heparin (subgroup 2) undergoing ART for in vitro
fertilisation/intracytoplasmic sperm injection were ana-
lysed. G-CSF was administered weekly once (34 Mill) in
11 patients, 38 patients received 2 9 13 Mill G-CSF per
week until the 12th week of gestation. Statistical analysis
was performed with SPSS for Windows (19.0), p \ 0.05
significant. The mean age of the study population was
37.3 ± 4.4 years (mean ± standard deviation) and dif-
fered not significantly between patients and subgroups.
However, the number of early miscarriages was signifi-
cantly higher in the G-CSF group as compared to the
subgroups (G-CSF 2.67 ± 1.27, subgroup 1 0.85 ± 0.91,
subgroup 2 0.64 ± 0.74) and RM patients receiving G-CSF
had significantly more often a late embryo transfer (day 5)
(G-CSF 36.7 %, subgroup 1 12.1 %, subgroup 2 8.9 %).
The LBR of patients and the subgroups differed signifi-
cantly (G-CSF 32 %, subgroup 1 13 %, subgroup 2 14 %).
Side effects were present in less than 10 % of patients,
consisting of irritation at the injection side, slight leuko-
cytosis, rise of the temperature (\38 °C), mild bone pain
and hyperemesis gravidarum. None of the newborn showed
any kind of malformations. According to our data, G-CSF
seems to be a safe and promising immunological treatment
option for RM patients. However, with regard to the ret-
rospective setting and the possible bias of a higher rate of
late embryo transfers in the G-CSF group additional studies
are needed to further strengthen our results.
Keywords Recurrent miscarriage
Assisted reproduction Immunological treatment
G-CSF
Brain-Derived Neurotrophic Factor in Neuroimmunology:
Lessons Learned from Multiple Sclerosis Patients
and Experimental Autoimmune Encephalomyelitis Models
Fred Lu¨ hder • Ralf Gold •
Alexander Flu¨ gel • Ralf A. Linker
Received: 22 June 2012 / Accepted: 20 December 2012 / Published online: 3 January 2013
Ó L. Hirszfeld Institute of Immunology and Experimental Therapy, Wroclaw, Poland 2013
Abstract The concept of neuroprotective autoimmunity
implies that immune cells, especially autoantigen-specific
T cells, infiltrate the central nervous system (CNS) after injury
and contribute to neuroregeneration and repair by secreting
soluble factors. Amongst others, neurotrophic factors and
neurotrophins such as brain-derived neurotropic factor
(BDNF) are considered to play an important role in this pro-
cess. New data raise the possibility that this concept could also
be extended to neuroinflammatory diseases such as multiple
sclerosis (MS) where autoantigen-specific T cells infiltrate the
CNS, causing axonal/neuronal damage on the one hand, but
also providing neuroprotective support on the other hand. In
this review, we summarize the current knowledge on BDNF
levels analyzed in MS patients in different compartments and
its correlation with clinical parameters. Furthermore, new
approaches in experimental animal models are discussed that
attempt to decipher the functional relevance of BDNF in
autoimmune demyelination.
Keywords Experimental autoimmune encephalomyelitis
(EAE) Neurotrophins Brain-derived neurotropic factor
(BDNF) Multiple sclerosis (MS)
Inflammation, Cytokines and Insulin Resistance:
A Clinical Perspective
Verena Wieser • Alexander R. Moschen •
Herbert Tilg
Received: 17 May 2012 / Accepted: 20 December 2012 / Published online: 10 January 2013
Ó L. Hirszfeld Institute of Immunology and Experimental Therapy, Wroclaw, Poland 2013
Abstract Obesity and obesity-related disorders have
dramatically increased globally in the last decades. These
entities are commonly associated with a state of insulin
resistance and the relationship between extensive lipid
deposition and insulin resistance is widely accepted. The
underlying mechanisms for insulin resistance, however, are
still incompletely understood. Evidence from experimental
research and human studies in the last years has suggested
that innate immune pathways and inflammatory mecha-
nisms also might play a role. Insulin resistance in case
of obesity is commonly accompanied by low-grade sys-
temic inflammation and adipose tissue inflammation. The
expression of various pro-inflammatory cytokines such as
tumor necrosis factor (TNF)-a, interleukin (IL)-1 and IL-6
is increased in adipose tissue and its expression linked to
systemic inflammation and accompanying insulin resis-
tance. Weight loss reduces this enhanced cytokine expres-
sion in the adipose tissue and thereby improves systemic
inflammation. Whereas there is also substantial evidence
that pro-inflammatory cytokines, certain members of the
inflammasome and various transcription factors such as
nuclear factor-jB play a major role in animal models of
insulin resistance and type 2 diabetes, human studies
neutralizing certain pro-inflammatory cytokines suggest so
far that not all pro-inflammatory cytokines are of equal
clinical importance. First clinical studies favor an impor-
tant role for IL-1 family members and probably IL-6 but
not for TNF-a in insulin-resistant states. Although we still
have missing links in the understanding of insulin resis-
tance, certain inflammatory pathways have also evolved in
humans as central players.
Keywords Innate immunity Type 2 diabetes
Adipocytokines Anti-cytokine therapies
Molecular Beacons: A Novel Optical Diagnostic Tool
Su-Xia Han • Xi Jia • Jin-lu Ma • Qing Zhu
Received: 3 April 2012 / Accepted: 20 December 2012 / Published online: 5 January 2013
Ó L. Hirszfeld Institute of Immunology and Experimental Therapy, Wroclaw, Poland 2012
Abstract As a result of the efforts of the Human Genome
Project and the rise in demand for molecular diagnostic
assays, the development and optimization of novel
hybridization probes have focused on speed, reliability, and
accuracy in the identification of nucleic acids. Molecular
beacons (MBs) are single-stranded, fluorophore-labeled
nucleic acid probes that are capable of generating a fluo-
rescent signal in the presence of target, but are dark in the
absence of target. Because of the high specificity and
sensitivity characteristics, MBs have been used in variety
of fields. In this review, MBs are introduced and discussed
as diagnostic tools in four sections: several technologies of
MBs will be illustrated primarily; the limitation of MBs
next; the third part is new fashions of MBs; and the last one
is to present the application of MBs in disease diagnosis.
Keywords Molecular beacons (MBs)
Nucleic acid probe Fluorescence resonance energy
transfer (FRET) Diagnosis
The Evolutionary Role of the IL-33/ST2 System
in Host Immune Defence
Susanne Sattler • Hermelijn H. Smits •
Damo Xu • Fang-Ping Huang
Received: 7 July 2012 / Accepted: 20 December 2012 / Published online: 3 January 2013
Ó L. Hirszfeld Institute of Immunology and Experimental Therapy, Wroclaw, Poland 2012
Abstract Interleukin (IL)-33 is a recently identified
pleiotropic cytokine, which can orchestrate complex innate
and adaptive immune responses in immunity and disease. It
has been characterized as a cytokine of the IL-1 family and
affects a wide range of immune cells by signalling through
its receptor ST2L. Accumulating evidence suggests a cru-
cial role of IL-33/ST2 in inducing and modifying host
immune responses against a variety of pathogens including
parasites, bacteria, viruses and fungi as well as sterile
insults of both endogenous and exogenous source. In this
review, we endeavour to give a comprehensive overview of
the current knowledge about the role of IL-33 and its
receptor ST2 in host defence against infections.
Keywords IL-33 ST2 Infection Infectious disease
Host defence Immune response
Associated Immunological Disorders and Cellular Immune
Dysfunction in Thymoma: A Study of 87 Cases from Thailand
Charat Thongprayoon • Pakpoom Tantrachoti •
Parkpoom Phatharacharukul • Supranee Buranapraditkun •
Jettanong Klaewsongkram
Received: 16 May 2012 / Accepted: 21 September 2012 / Published online: 11 December 2012
Ó L. Hirszfeld Institute of Immunology and Experimental Therapy, Wroclaw, Poland 2012
Abstract Several immune disorders are often associated
with thymoma. The aim of this study was to analyze the
correlation between clinicopathological features of Thai
patients with thymoma and concomitant immune-mediated
diseases. Medical records of 87 patients diagnosed with
thymoma during a 10-year period were retrospectively
reviewed. Peripheral blood T cell subsets along with
cytokine responses in 15 thymoma patients and 15 healthy
controls were comparatively analyzed. The results dem-
onstrated that thymoma type AB and B2 were the most
common types among patients diagnosed with thymoma.
The most common presentation was incidentaloma, fol-
lowed by local chest symptoms and autoimmune diseases.
The prevalence of autoimmune diseases, immunodefi-
ciency states, and secondary neoplasms was 34.5, 10.3, and
10.3 %, respectively. Autoimmune diseases were most
frequently found in thymoma type B2 and sometimes
associated with clinical immunodeficiency, although clas-
sic Good’s syndrome was rare. Patients with thymoma had
significantly lower percentage CD4?ve T cells and inter-
feron c response, but higher percentage regulatory T cells
than those in healthy controls. This study indicated that the
aberrant immunologic disorders comprising autoimmune
diseases, immunodeficiency states, and secondary neo-
plasms were found in almost 40 % of Thai patients with
thymoma and possibly related to defectiva cytokine
responses and altered T cell subsets.
Keywords Thymoma Paraneoplastic syndromes
Autoimmunity Immunologic deficiency syndrome
Neoplasms
Recombinant k Bacteriophage Displaying Nanobody
towards Third Domain of HER-2 Epitope Inhibits
Proliferation of Breast Carcinoma SKBR-3 Cell Line
Alireza Shoae-Hassani • Seyed Abdolreza Mortazavi-Tabatabaei •
Shiva Sharif • Shabnam Madadi • Hamidreza Rezaei-Khaligh • Javad Verdi
Received: 22 February 2012 / Accepted: 20 August 2012 / Published online: 7 December 2012
Ó L. Hirszfeld Institute of Immunology and Experimental Therapy, Wroclaw, Poland 2012
Abstract Phage display of many nanobodies via fila-
mentous phage in combination with helper phage has been
reported by many scientists. The aim of this study was to
produce lambda (k) bacteriophage displaying high-affinity
nanobody against HER-2 expressing breast carcinoma
cells. Bacteriophage k is a temperate phage with inherent
biological safety in mammalian cells. Here we report the
construction of a recombinant k phage that efficiently
expresses specific nanobody towards third domain of HER-
2 target on SKBR-3 and MCF-7 cell lines in vitro. We
constructed recombinant k phage particles containing a
mammalian expression cassette, C-Myc tagged, encoding
VHH gene of camelid anti HER-2 third domain epitope
using k ZAP-cytomegalic virus (CMV) vector. The SKBR-3,
MCF-7 and human endometrial stem cells were treated by
the nanobody displayed recombinant k phage. The cell
growth inhibition assay was performed by MTT Cell
Viability Assay Kit. After the fourth round of biopanning
there was a significant enrichment in the phage specifically
binding to the antigen. The ratio of targeted phage
increased approximately 1,000-fold in the fifth round. The
nanobody expressed by k ZAP-CMV-VHH phagemid
cloned in k bioparticles significantly inhibited the prolif-
eration of HER-2 positive SKBR-3 and MCF-7 cells.
Recombinant bacteriophage k ZAP-CMV-VHH-cDNA
could be used efficiently for construction of nanobodies to
mortify HER-2 positive breast carcinoma cells as a nano-
medical therapeutic.
Keywords Nanomedicine Nanobody Nanobioparticle
Recombinant
Autophagy, Apoptosis, Mitoptosis and Necrosis: Interdependence
Between Those Pathways and Effects on Cancer
Wiem Chaabane • Sırma D. User • Mohamed El-Gazzah • Roman Jaksik •
Elaheh Sajjadi • Joanna Rzeszowska-Wolny • Marek J. Łos
Received: 22 May 2012 / Accepted: 6 September 2012 / Published online: 11 December 2012
Ó L. Hirszfeld Institute of Immunology and Experimental Therapy, Wroclaw, Poland 2012
Abstract Cell death is a fundamental ingredient of life.
Thus, not surprisingly more than one form of cell death
exists. Several excellent reviews on various forms of cell
death have already been published but manuscripts
describing interconnection and interdependence between
such processes are uncommon. Here, what follows is a brief
introduction on all three classical forms of cell death, fol-
lowed by a more detailed insight into the role of p53, the
master regulator of apoptosis, and other forms of cell death.
While discussing p53 and also the role of caspases in cell
death forms, we offer insight into the interplay between
autophagy and apoptosis, or necrosis, where autophagy may
initially serve pro-survival functions. The review moves
further to present some details about less researched forms of
programmed cell death, namely necroptosis, necrosis and
mitoptosis. These ‘‘mixed’’ forms of cell death allow us to
highlight the interconnected nature of cell death forms,
particularly apoptosis and necrosis. The interdependence
between apoptosis, autophagy and necrosis, and their sig-
nificance for cancer development and treatment are also
analyzed in further parts of the review. In the concluding
parts, the afore-mentioned issues will be put in perspective
for the development of novel anti-cancer therapies.
Keywords AMPK Hsp-70 Mdm2 Mitochondria
mTOR RIPK PARP-1 PKC-delta TNF VHL
While at Rome miRNA and TRAIL Do Whatever BCR-ABL
Commands to Do
Ammad Ahmad Farooqi • Ali Nawaz • Zeeshan Javed •
Shahzad Bhatti • Muhammad Ismail
Received: 26 January 2012 / Accepted: 20 August 2012 / Published online: 11 December 2012
Ó L. Hirszfeld Institute of Immunology and Experimental Therapy, Wroclaw, Poland 2012
Abstract It is a well-acclaimed fact that proteins
expressed as a consequence of oncogenic fusions, muta-
tions or amplifications can facilitate ectopic protein–
protein interactions that re-wire signal dissemination
pathways, in a manner that escalates malignancy. BCR-
ABL-mediated signal transduction cascades in leukemic
cells are assembled and modulated by a finely controlled
network of protein–protein interactions, mediated by
characteristic signaling domains and their respective
binding motifs. BCR-ABL functions in a cell context-
specific and cell type-specific manner to integrate signals
that affect uncontrolled cellular proliferation. In this
review, we draw attention to the recent progress made in
outlining resistance against TRAIL-mediated apoptosis and
diametrically opposed roles of miRNAs in BCR-ABL-
positive leukemic cells. BCR-ABL governs carcinogenesis
through well-organized web of antiapoptotic proteins and
over-expressed oncomirs which target death receptors and
pro-apoptotic genes. Set of oncomirs which inversely cor-
relate with expression of TRAIL via suppression of SMAD
is an important dimension which is gradually gaining
attention of the researchers. Contrary to this, some current
findings show a new role of BCR-ABL in nucleus with
spotlight on apoptosis. It seems obvious that genetic het-
erogeneity of leukemias poses therapeutic challenges, and
pharmacological agents that target components of the
cancer promoting nano-machinery still need broad experi-
mental validation to be considered competent as a
component of the therapeutic arsenal for this group of
diseases. Rapidly developing technologies are empowering
us to explain the molecular ‘‘nature’’ of a patient and/or
tumor and with this integration of personalized medicine,
with maximized efficacy, cost effectiveness will hopefully
improve survival chances of the patient.
Keywords BCR-ABL TRAIL miRNA Apoptosis
Cancer
Glioblastoma, a Brief Review of History, Molecular Genetics,
Animal Models and Novel Therapeutic Strategies
Sameer Agnihotri • Kelly E. Burrell • Amparo Wolf •
Sharzhad Jalali • Cynthia Hawkins • James T. Rutka •
Gelareh Zadeh
Received: 30 May 2012 / Accepted: 22 November 2012 / Published online: 7 December 2012
Ó L. Hirszfeld Institute of Immunology and Experimental Therapy, Wroclaw, Poland 2012
Abstract Glioblastoma (GBM) is the most common and
lethal primary brain tumor. Over the past few years tre-
mendous genomic and proteomic characterization along
with robust animal models of GBM have provided
invaluable data that show that ‘‘GBM’’, although histo-
logically indistinguishable from one another, are comprised
of molecularly heterogenous diseases. In addition, robust
pre-clinical models and a better understanding of the core
pathways disrupted in GBM are providing a renewed
optimism for novel strategies targeting these devastating
tumors. Here, we summarize a brief history of the disease,
our current molecular knowledge, lessons from animal
models and emerging concepts of angiogenesis, invasion,
and metabolism in GBM that may lend themselves to
therapeutic targeting.
Keywords Glioblastoma Molecular genetics of glioma
Mouse models Novel molecular targets
Regulation of Atherogenesis by Chemokines
and Chemokine Receptors
Wuzhou Wan • Philip M. Murphy
Received: 4 June 2012 / Accepted: 18 November 2012 / Published online: 7 December 2012
Ó L. Hirszfeld Institute of Immunology and Experimental Therapy, Wroclaw, Poland 2012
Abstract Atherosclerosis is a chronic inflammatory and
metabolic disorder affecting large- and medium-sized
arteries, and the leading cause of mortality worldwide. The
pathogenesis of atherosclerosis involves accumulation of
lipids and leukocytes in the intima of blood vessel walls
creating plaque. How leukocytes accumulate in plaque
remains poorly understood; however, chemokines acting at
specific G protein-coupled receptors appear to be impor-
tant. Studies using knockout mice suggest that chemokine
receptor signaling may either promote or inhibit athero-
genesis, depending on the receptor. These proof of concept
studies have spurred efforts to develop drugs targeting the
chemokine system in atherosclerosis, and several have
shown beneficial effects in animal models. This study will
review key discoveries in basic and translational research
in this area.
Keywords Atherosclerosis Cardiology Immunology
Inflammation Antagonist
Professor Józef Lisowski (1928–2012)
The Member of AITE Advisory Board 1968–1997
New European Rules Regarding the Approval of Clinical Trials,
the Role of Ethics Committees and the Protection of Subjects
Herman Nys
Function of Junctional Adhesion Molecules (JAMs)
in Leukocyte Migration and Homeostasis
Marie-Laure Arcangeli • Vincent Frontera •
Michel Aurrand-Lions
Received: 23 February 2012 / Accepted: 23 July 2012 / Published online: 1 September 2012
Ó L. Hirszfeld Institute of Immunology and Experimental Therapy, Wroclaw, Poland 2012
Abstract Homeostasis is a word widely used in the
scientific community to refer to the property of a system to
maintain its uniformity and functionality. In living organ-
isms, the word refers to the concept enunciated 150 years
ago by C. Bernard by which external variations must be
compensated for in order to maintain internal conditions
compatible with life. This is especially true in the case of
highly dynamic system such as the hematopoietic system
that requires the coordinated control of cell proliferation
and death within specialized microenvironments that are
anatomically distinct. As a consequence, hematopoietic
cell adhesion and migration must be tightly controlled in
order for hematopoietic cells to reach and to be maintained
in appropriate microenvironments. The junctional adhesion
molecules (JAMs) are adhesion molecules that belong to
the immunoglobulin superfamily (IgSf) and that have been
initially identified as important players controlling vascular
permeability and leukocyte transendothelial migration.
This involves the regulated localization of the JAMs at
lateral endothelial cell/cell borders and their interaction
with leukocyte integrins. More recently, some of the JAM
family members have also been found to be expressed by
stromal cells and to regulate chemokine secretion within
lymphoid organs, acting not only on leukocyte transendo-
thelial migration, but also on hematopoietic cell retention
within specialized microenvironments. This review sum-
marizes recent progress in understanding the role of the
JAMs in leukocyte adhesion and migration to tentatively
draw an integrated view of the homeostatic function of the
JAMs within the hematopoietic system.
Keywords Transendothelial migration Lymph node
Bone marrow Chemokine secretion
Generation of Functional T-Regulatory Cells
in Children with Metabolic Syndrome
Włodzimierz Łuczyn´ ski • Natalia Wawrusiewicz-Kurylonek •
El_zbieta Iłendo • Artur Bossowski • Barbara Głowin´ ska-Olszewska •
Adam Kre˛towski • Anna Stasiak-Barmuta
Received: 5 April 2012 / Accepted: 23 July 2012 / Published online: 7 October 2012
Ó L. Hirszfeld Institute of Immunology and Experimental Therapy, Wroclaw, Poland 2012
Abstract Recent research implies a role of decreased
number and/or function of T-regulatory cells (Tregs) in
low-grade inflammation associated with obesity and
atherosclerosis. The enhancement of atheroprotective
immunity by the expansion of Tregs could serve as a
therapeutic strategy in obesity-related immunological
disturbances. The aim of our study was an attempt to
generate Treg cells in children with risk factors for the
development of cardiovascular disease and to compare
the results to those obtained in healthy subjects. The
study group consisted of 30 children with metabolic
syndrome (MS) and 30 controls. Conventional
CD4?CD25– cells separated from the peripheral blood
were converted into Treg cells with the use of CD3/
CD28 antibodies and interleukin (IL)-2/transforming
growth factor (TGF)-b stimulation. The expression of
critical Treg molecules and cytokines was assessed at
mRNA and protein levels. The percentages of Treg cells
in the peripheral blood were significantly lower in the
children with MS compared to the healthy subjects. After
the culture with CD3/CD28 and IL-2/TGF-b we detected
a significant increase in the expression of Tregs marker
transcription factor FoxP3. The Tregs induced from the
children with MS varied from the ones obtained in the
controls in the expression of some molecules at mRNA
level (e.g. IL-27, LGAL, KLF10 and NRP1) yet not in
proliferation studies. For the first time, we have dem-
onstrated the possibility of generating functional Treg
cells in children with MS. The results of our study could
be used in the design of therapeutic interventions in
obesity associated immunologic disturbances.
Keywords Children Immunology
Metabolic syndrome Obesity
Serum Tumor Necrosis Factor-a and Interleukin-10 Levels
as Markers to Predict Outcome of Patients with Chronic
Lymphocytic Leukemia in Different Risk Groups Defined
by the IGHV Mutation Status
Ewa Lech-Maranda • Olga Grzybowska-Izydorczyk • Krystyna Wyka • Wojciech Mlynarski •
Maciej Borowiec • Karolina Antosik • Barbara Cebula-Obrzut • Hanna Makuch-Lasica • Grazyna Nowak •
Gabriela Klimkiewicz-Wojciechowska • Ewa Wawrzyniak • Przemyslaw Bilinski • Tadeusz Robak •
Krzysztof Warzocha
Received: 26 March 2012 / Accepted: 23 July 2012 / Published online: 4 September 2012
Ó L. Hirszfeld Institute of Immunology and Experimental Therapy, Wroclaw, Poland 2012
Abstract Tumor necrosis factor (TNF)-a and interleukin
(IL)-10 are cytokines involved in the balance between cell-
mediated and humoral immunity. We investigated whether
serum TNF-a and IL-10 levels have any impact on clinical
outcome of patients with chronic lymphocytic leukemia
(CLL). TNF-a and IL-10 levels were determined in the
serum of 160 CLL patients at the time of diagnosis. The
cytokine low-risk group consisted of patients with either
TNF-a and IL-10 levels below their medians or those with
only one elevated parameter. Both TNF-a and IL-10 levels
greater than or equal to their medians defined the cytokine
high-risk group. The high-risk patients presented a shorter
3-year treatment-free survival (TFS) than low-risk subjects
(15 vs. 69.6 %; p \ 0.0001). The high-risk group
(p = 0.0002) along with high leukocyte count (p \ 0.0001)
and unmutated immunoglobulin heavy-chain variable
region genes (p \ 0.0001) independently predict the risk of
progression in patients with Rai stage 0–II. Furthermore,
the high-risk group had an independent prognostic impact
on shorter TFS both in patients with mutated (24.3 vs.
78.2 %; p \ 0.0001) and unmutated (8.2 vs. 49 %;
p = 0.004) immunoglobulin heavy-chain variable region
genes (IGHV) as compared to the low-risk group. The
estimated 5-year overall survival (OS) of high-risk patients
was shorter than those in the low-risk group (83.3 vs.
97.1 %; p = 0.003). Multivariate analysis demonstrated
the cytokine high-risk group (p = 0.02) followed by Rai
stage III–IV (p = 0.048) to be independent factors predict-
ing shorter OS. At diagnosis, TNF-a and IL-10 may predict the
outcome of patients with CLL.
Keywords Chronic lymphocytic leukemia
Tumor necrosis factor-a Interleukin-10
Immunoglobulin heavy-chain variable region genes
Outcome
In Vitro Invasion and Survival of Porphyromonas gingivalis
in Gingival Fibroblasts; Role of the Capsule
Muhammad Irshad • Wil A. van der Reijden •
Wim Crielaard • Marja L. Laine
Received: 21 March 2012 / Accepted: 23 July 2012 / Published online: 5 September 2012
Ó L. Hirszfeld Institute of Immunology and Experimental Therapy, Wroclaw, Poland 2012
Abstract Porphyromonas gingivalis is a Gram-negative,
anaerobic bacterium involved in periodontitis and peri-
implantitis that can invade and survive inside host cells in
vitro. P. gingivalis can invade human gingival fibroblasts
(GF), but no data are available about the role of P. gingi-
valis’ capsule in GF invasion. In the current study, we
aimed to determine the ability of three strains of P. gin-
givalis (encapsulated wild type W83, non-encapsulated
HG91 and the non-encapsulated insertional isogenic
knockout mutant of W83, DEpsC) to invade GF and the
ability of internalized P. gingivalis to survive in vitro
antibiotic treatment. The ability of P. gingivalis strains to
invade GF was tested using an antibiotic protection assay at
multiplicity of infection (MOI) 100 and 1000. The survival
of internalized P. gingivalis cells was further analyzed by
subsequent in vitro treatment with either metronidazole or
amoxicillin alone or a combination of metronidazole and
amoxicillin and anaerobic culture viability counts. All
strains of P. gingivalis used in this study were able to
invade GFs. The non-encapsulated mutant of W83 (DEpsC
mutant) was significantly more invasive than the wild type
W83 at MOI 100 (p value 0.025) and MOI 1000 (p value
0.038). Furthermore, internalized P. gingivalis was able to
resist in vitro antibiotic treatment. As demonstrated by
the differences in invasion efficiencies of P. gingivalis
strain W83 and its isogenic mutant DEpsC, the capsule of
P. gingivalis makes it less efficient in invading gingival
fibroblasts. Moreover, internalized P. gingivalis can sur-
vive antibiotic treatment in vitro.
Keywords Periodontitis
Mechanism of antibiotic resistance Internalization
Capsule
Putative Biological Mechanisms of Efficiency of Substrate
Reduction Therapies for Mucopolysaccharidoses
Zyta Banecka-Majkutewicz • Joanna Jako´bkiewicz-Banecka •
Magdalena Gabig-Cimin´ ska • Alicja We˛grzyn •
Grzegorz We˛grzyn
Received: 28 February 2012 / Accepted: 15 June 2012 / Published online: 5 September 2012
Ó L. Hirszfeld Institute of Immunology and Experimental Therapy, Wroclaw, Poland 2012
Abstract Mucopolysaccharidoses (MPS) are inherited
metabolic diseases caused by mutations in genes coding for
lysosomal enzymes involved in the degradation of gly-
cosaminoglycans (GAGs). Dysfunction of any of these
enzymes results in the accumulation of GAGs, which leads
to severe clinical symptoms and significantly shortened life
span. Several kinds of therapies have been proposed to treat
MPS, including bone marrow or stem cell transplantation,
enzyme replacement therapy, and gene therapy. Another
option is substrate reduction therapy (SRT), in which
synthesis of GAGs is inhibited. Recent studies employing
in vitro and animal models suggested that this therapy may
be efficient in decreasing levels of GAGs in MPS cells,
including those bearing two null alleles of the affected
gene. Results of behavioral tests in animals as well as some
preliminary clinical observations with pediatric patients
corroborated the suggestions about possible efficacy of
SRT in MPS treatment, including brain functions. Efficient
reduction of GAG levels in MPS cells homozygous for null
mutations may be intriguing in the commonly accepted
scheme of SRT mode of action. In this paper, we propose
an explanation of this phenomenon, based on already
known facts. Thus, we suggest that SRT may lead to
reduction of GAG levels in MPS cells due to inhibition of
efficiency of GAG synthesis combined with (a) any read-
through of the stop codon, (b) dilution of already
accumulated GAGs due to cell growth followed by cell
divisions, and (c) action of endoglycosidases degrading
GAGs, e.g., heparanase, in combination with functional
GAG-specific hydrolases.
Keywords Lysosomal storage diseases
Mucopolysaccharidoses Substrate reduction therapy
Gene expression-targeted isoflavone therapy Genistein
Glycosaminoglycans
The Lnk Adaptor Protein: A Key Regulator of Normal
and Pathological Hematopoiesis
Laura Velazquez
Received: 15 February 2012 / Accepted: 6 August 2012 / Published online: 19 September 2012
Ó L. Hirszfeld Institute of Immunology and Experimental Therapy, Wroclaw, Poland 2012
Abstract The development and function of blood cells
are regulated by specific growth factors/cytokines and their
receptors’ signaling pathways. In this way, these factors
influence cell survival, proliferation and differentiation of
hematopoietic cells. Central to this positive and/or negative
control are the adaptor proteins. Since their identification
10 years ago, members of the Lnk adaptor protein family
have proved to be important activators and/or inhibitors in
the hematopoietic, immune and vascular system. In par-
ticular, the generation of animal and cellular models for the
Lnk and APS proteins has helped establish the physiolog-
ical role of these molecules through the identification of
their specific signaling pathways and the characterization
of their binding partners. Moreover, the recent identifica-
tion of mutations in the LNK gene in myeloproliferative
disorders, as well as the correlation of a single nucleotide
polymorphism on LNK with hematological, immune and
vascular diseases have suggested its involvement in the
pathophysiology of these malignancies. The latter findings
have thus raised the possibility of addressing Lnk signaling
for the treatment of certain human diseases. This review
therefore describes the pathophysiological role of this
adaptor protein in hematological malignancies and the
potential benefits of Lnk therapeutic targeting.
Keywords Adaptor protein Cytokine signaling
Regulation Hematopoietic cells Human diseases
Animal models
Platelets in Inflammation and Immune Modulations:
Functions Beyond Hemostasis
Hsuan-Shun Huang • Hsin-Hou Chang
Received: 4 September 2011 / Accepted: 29 February 2012 / Published online: 1 September 2012
Ó L. Hirszfeld Institute of Immunology and Experimental Therapy, Wroclaw, Poland 2012
Abstract Platelets play central roles for maintaining the
homeostasis of the blood coagulation. As they are also
involved in immune responses and host defenses, increas-
ing evidences have suggested that platelets exert other roles
beyond their well-recognized function in preventing
bleeding. This review is focused on inflammation, allergy
and immune modulations of platelets. Platelets conduct
immunoregulation through secretion of functional media-
tors, interaction with various immune cells, endothelial
cells and beneficial for the leukocyte infiltration to
inflamed/allergic tissues. In these regulations, the leuko-
cytes are influenced by and receiving the signals from
platelets. In contrast, rare attentions were focused on
platelet regulations by immune system. An intriguingly
example in the intravenous immunoglobulin (IVIg) treat-
ment is discussed, in which dendritic cells exert anti-
inflammatory effect through platelets. This further suggests
that coagulant and immune systems are tightly associated
rather than separate entities. The cross-talks between these
two systems implicate that platelet therapy may have
application beyond thrombosis, and immune interventions
may have potentials to treat thrombosis diseases.
Keywords Platelet Dendritic cell Immune regulation
Intravenous immunoglobulin
Pathophysiological Mechanisms of Carotid Plaque Vulnerability:
Impact on Ischemic Stroke
Jaroslav Pelisek • Hans-Henning Eckstein •
Alma Zernecke
Received: 5 April 2012 / Accepted: 6 August 2012 / Published online: 4 September 2012
Ó L. Hirszfeld Institute of Immunology and Experimental Therapy, Wroclaw, Poland 2012
Abstract Stroke is among the major causes of mortality
and disabilities in the world. About 80 % of all strokes in
the anterior circulation are ischemic and up to 20 % of all
ischemic strokes are caused by extracranial atherosclerotic
carotid artery stenosis. The prevalence of a cervical inter-
nal carotid artery stenosis increases with age and can be
found in 6.9 % of the elderly population ([65 years).
Atherosclerotic changes of the carotid vessel wall can lead
to plaque vulnerability and may result in arterio-arterial
embolism, which frequently underlie carotid-related cere-
brovascular ischemic events. Carotid atherosclerosis is
characterised by inflammation, extensive degradation of
extracellular matrix components, neovascularization, and
as recently recognised is also affected by epigenetic
changes. These factors accelerate the progression of ath-
erosclerosis towards vulnerable plaques and increase the
risk of ischemic stroke. In this review, the main patho-
physiological mechanisms leading to rupture-prone carotid
artery plaques and successive ischemic stroke are consid-
ered. It is important to recognise the heterogeneity of
atherosclerosis and that various pathophysiological pro-
cesses dissected in this review are not acting individually,
but rather in a complementary manner. The identification
and careful integration of all relevant factors will be
required for the development of future diagnostic and
therapeutic strategies.
Keywords Carotid atherosclerosis
Vulnerable carotid plaque Ischemic stroke
Aberrant Function and Differentiation of Monocytes in End Stage
Renal Disease
Krzysztof Laudan´ ski • Zbigniew Nowak
Received: 19 April 2012 / Accepted: 23 July 2012 / Published online: 19 October 2012
Ó L. Hirszfeld Institute of Immunology and Experimental Therapy, Wroclaw, Poland 2012
Abstract Patients with end stage renal disease (ESRD)
suffer from many disturbances of the immune system.
These immunopathologies are related to the higher failure
of vaccination, and increased prevalence of infections and
neoplasms. In the presented article, we review the current
data regarding the role of monocytes in immune dysfunc-
tions which are observed in terminal renal failure. As
monocytes play a pivotal role in regulating the function of
the immune system, their dysfunction can have a profound
effect on the immune system and may lead to accelerated
arteriosclerosis and deteriorating overall health conditions.
More specifically, we suggest that peripheral blood
monocytes in patients with ESRD are chronically activated,
and their functional and phenotypical features resemble
those of inflammatory macrophages. This state of chronic
inflammation is unfavorable for dendritic cells and conse-
quently, the prevalence of dendritic cells is reduced. As
these effects are consistent across different modes of
dialysis, they are probably mediated by the uremia itself.
Keywords End stage renal disease Monocyte
Dendritic cells Inflammation
Immunological Functions of Steryl Glycosides
Michio Shimamura
Received: 20 January 2012 / Accepted: 8 August 2012 / Published online: 31 August 2012
Ó L. Hirszfeld Institute of Immunology and Experimental Therapy, Wroclaw, Poland 2012
Abstract Steryl glycosides, sterols glycosylated at the
3b-hydroxy group, have been widely found in plants, algae,
and fungi, but are rare in bacteria and animals. Glycosyl-
ation of sterols is known to modify properties of the cell
membrane and confer resistance against stresses by freez-
ing or heat-shock on cells. Furthermore, accumulating
evidence obtained from recent research suggests important
biological functions of steryl glycosides, including regu-
lation of host defenses against pathogens, lipid metabolism,
and developmental events. This review is focused on the
immunological functions of steryl glycosides, such as
modulation of host immune functions upon exposure to
cholesteryl glycosides produced by pathogenic bacteria.
Keywords Steryl glycoside Invariant NKT cell
Helicobacter pylori Immunomodulation
Immunogenicity to Biologics: Mechanisms, Prediction
and Reduction
Swaminathan Sethu • Karthik Govindappa •
Mohammad Alhaidari • Munir Pirmohamed •
Kevin Park • Jean Sathish
Received: 9 January 2012 / Accepted: 11 May 2012 / Published online: 29 August 2012
Ó L. Hirszfeld Institute of Immunology and Experimental Therapy, Wroclaw, Poland 2012
Abstract Currently, there is a significant rise in the
development and clinical use of a unique class of pharma-
ceuticals termed as Biopharmaceuticals or Biologics, in the
management of a range of disease conditions with, remark-
able therapeutic benefits. However, there is an equally
growing concern regarding development of adverse effects
like immunogenicity in the form of anti-drug antibodies
(ADA) production and hypersensitivity. Immunogenicity to
biologics represents a significant hurdle in the continuing
therapy of patients in a number of disease settings. Efforts
focussed on the identification of factors that contribute
towards the onset of immunogenic response to biologics
have led to reductions in the incidence of immunogenicity.
An in-depth understanding of the cellular and molecular
mechanism underpinning immunogenic responses will
likely improve the safety profile of biologics. This review
addresses the mechanistic basis of ADA generation to biol-
ogics, with emphasis on the role of antigen processing and
presentation in this process. The article also addresses the
potential contribution of complement system in augmenting
or modulating this response. Identifying specific factors that
influences processing and presentation of biologic-derived
antigens in different genotype and disease background may
offer additional options for intervention in the immunogenic
process and consequently, the management of immunoge-
nicity to biologics.
Keywords Biologics Immunogenicity
Antigen processing Complement
Protein Kinase Cd in Apoptosis: A Brief Overview
Meng Zhao • Li Xia • Guo-Qiang Chen
Received: 4 June 2011 / Accepted: 6 August 2012 / Published online: 24 August 2012
Ó L. Hirszfeld Institute of Immunology and Experimental Therapy, Wroclaw, Poland 2012
Abstract Protein kinase C-delta (PKCd), a member of
the lipid-regulated serine/threonine PKC family, has been
implicated in a wide range of important cellular processes.
In the past decade, the critical role of PKCd in the regu-
lation of both intrinsic and extrinsic apoptosis pathways
has been widely explored. In most cases, over-expression
or activation of PKCd results in the induction of apoptosis.
The phosphorylations and multiple cell organelle translo-
cations of PKCd initiate apoptosis by targeting multiple
downstream effectors. During apoptosis, PKCd is proteo-
lytically cleaved by caspase-3 to generate a constitutively
activated catalytic fragment, which amplifies apoptosis
cascades in nucleus and mitochondria. However, PKCd
also exerts its anti-apoptotic and pro-survival roles in some
cases. Therefore, the complicated role of PKCd in apop-
tosis appears to be stimulus and cell type dependent. This
review is mainly focused on how PKCd gets activated in
diverse ways in response to apoptotic signals and how
PKCd targets different downstream regulators to sponsor or
restrain apoptosis induction.
Keywords Protein kinase C-delta Apoptosis
Signaling molecules
Effect of the Proline-Rich Polypeptide Complex/ColostrininTM
on the Enzymatic Antioxidant System
Agnieszka Zabłocka • Maria Janusz
Received: 10 November 2011 / Accepted: 20 April 2012 / Published online: 28 August 2012
Ó L. Hirszfeld Institute of Immunology and Experimental Therapy, Wroclaw, Poland 2012
Abstract Proline-rich polypeptide complex (PRP) and its
constituent nonapeptide (NP) possess immunoregulatory and
procognitive properties. PRP in the form of sublingually
administered tablets called ColostrininTM improves the
outcome of patients with Alzheimer’s disease (AD). Free
radical-induced oxidative stress has been implicated in the
pathogenesis of AD. It has been previously shown that PRP
and NP inhibit overproduction of reactive oxygen species,
nitric oxide and proinflammatory cytokines induced by
lipopolysaccharide or PMA. Antioxidant defense includes
both low molecular weight components and enzymatic
systems including dismutases, catalase, glutathione reduc-
tase (GSSGR) and glutathione peroxidase (GSHPx). An
early event during the development of AD is lipid and
protein peroxidation. PRP and NP showed no modulatory
effect on lipid peroxidation. A protective effect on protein
oxidation was found only when high doses of NP were used.
We have previously shown, in a model of human peripheral
blood mononuclear cells, that PRP/NP affects activities of
superoxide dismutase and NF-jB. In the present study with
the use of the same cell model and whole blood cells we
observed an activatory effect of PRP/NP on GSHPx and
GSSGR activity but not catalase. The observed effect sug-
gests that PRP/NP can act as a modulatory agent of the ‘‘first
line’’ of antioxidant defense. It can be assumed therefore that
PRP/Colostrinin by regulation of the early phase of the
redox system does not reduce but rather prevents oxidative
damage. This effect may shed some light on the beneficial
effect of PRP/Colostrinin in AD patients.
Keywords Proline-rich polypeptide complex (PRP)
Human peripheral blood mononuclear cells (PBMC)
Oxidative stress Catalase Glutathione Glutathione
reductase Lipid and protein oxidation
Toll-Like Receptors Expression and NF-jB Activation
in Peritoneal Leukocytes in Morphine-Mediated Impairment
of Zymosan-Induced Peritonitis in Swiss Mice
Ewa Wypasek • Joanna Natorska •
Agnieszka Irena Mazur • El_zbieta Kołaczkowska
Received: 15 July 2011 / Accepted: 28 May 2012 / Published online: 23 August 2012
Ó L. Hirszfeld Institute of Immunology and Experimental Therapy, Wroclaw, Poland 2012
Abstract Zymosan-induced peritonitis represents a well-
described model of acute inflammation. The binding of
zymosan with its specific Toll-like receptors (TLR2 and
TLR6) on leukocytes initiates activation and phosphory-
lation of nuclear factor (NF)-jB, which leads to
accumulation of NF-jB p65 subunits in the nucleus and
subsequently up-regulation of the proinflammatory cyto-
kine genes expression. Intraperitoneal co-administration of
zymosan and morphine significantly inhibits peritonitis in
several strains of mice by decreasing the influx of exuda-
tory cells; however, mechanisms of this action still remain
unclear. We aimed to verify the effects of morphine on
NF-jB and TLRs expression at messenger RNA and pro-
tein levels during the early stages of zymosan-induced
peritonitis. Peritonitis was induced by a single injection of
zymosan A or zymosan supplemented with morphine in
Swiss mice. At selected time points, after stimulation,
peritoneal leukocytes were harvested. The TLRs and
NF-jB expression was assessed by real-time PCR and flow
cytometry. In comparison with the mice injected with
zymosan only, morphine co-injection significantly
decreased the expression of phospho-NF-jB and TLR2
in all investigated immunocompetent cells as well as
up-regulated the levels of nitric oxide (NO) in peritoneal
fluid. Moreover, supplementation of zymosan with mor-
phine altered the TLR, NF-jB and some proinflammatory
cytokines (keratinocyte-derived chemokine, tumor necrosis
factor-a) gene expression during ongoing inflammation.
We may postulate that after morphine stimulation perito-
neal leukocytes recognize less effectively zymosan
antigens because of impaired TLRs expression. The lower
TLR expression attenuates TLR-mediated signal transduc-
tion, which prevents NF-jB activation. Additionally, during
zymosan-induced peritonitis, morphine may modulate the
NF-jB expression, at least partially, by an up-regulated
release of NO, as suggested by others.
Keywords Peritonitis NF-jB TLR Morphine
Mice
Adaptive Immune Responses Associated with Breast Cancer
Relapse
Kyle K. Payne • Masoud H. Manjili
Received: 6 February 2012 / Accepted: 28 May 2012 / Published online: 22 August 2012
Ó L. Hirszfeld Institute of Immunology and Experimental Therapy, Wroclaw, Poland 2012
Abstract The generation, survival, and differentiation of
breast cancer stem cells (BCSC) in immunocompetent hosts
remain elusive. Some investigators have shown that BCSC
can be induced from epithelial tumor cells by the pathologic
epithelial to mesenchymal transition (EMT). Emerging evi-
dence suggests that the induction of EMT among epithelial
tumor cells originates from signals produced by the non-tumor
cells that constitute the tumor microenvironment, including
the immune effectors that infiltrate the tumors. Thus, this
suggests that the immune system not only has anti-tumor
function, but also paradoxically immunoedits tumors, facili-
tating tumor escape and progression. Indeed, many studies in
human breast cancers show both positive and negative asso-
ciations between the infiltration of various immune effectors
(e.g., CD4 and CD8 T cells) and the propensity to relapse with
metastatic disease. These observations suggest that distinct
types of immune effector cells may induce or inhibit tumor
relapse. This review focuses on recent advances in identifying
components of the immune system that may directly induce
tumor escape and relapse. We propose that levels of interferon
(IFN)-c production or levels of the expression of IFN-c
receptor a on tumor cells may determine whether tumor
inhibitory or relapse-promoting effect of IFN-c may prevail.
Keywords Breast cancer stem cells Relapse
Her-2/neu IFN-c Immunoediting Tumor escape
Hepatitis C Virus-Mediated Modulation of Cellular Immunity
Erwin Daniel Brenndo¨rfer • Matti Sa¨llberg
Received: 20 October 2011 / Accepted: 9 March 2012 / Published online: 22 August 2012
Ó L. Hirszfeld Institute of Immunology and Experimental Therapy, Wroclaw, Poland 2012
Abstract The hepatitis C virus (HCV) is a major cause of
chronic liver disease globally. A chronic infection can
result in liver fibrosis, liver cirrhosis, hepatocellular car-
cinoma and liver failure in a significant ratio of the
patients. About 170 million people are currently infected
with HCV. Since 80 % of the infected patients develop a
chronic infection, HCV has evolved sophisticated escape
strategies to evade both the innate and the adaptive
immune system. Thus, chronic hepatitis C is characterized
by perturbations in the number, subset composition and/or
functionality of natural killer cells, natural killer T cells,
dendritic cells, macrophages and T cells. The balance
between HCV-induced immune evasion and the antiviral
immune response results in chronic liver inflammation and
consequent immune-mediated liver injury. This review
summarizes our current understanding of the HCV-medi-
ated interference with cellular immunity and of the factors
resulting in HCV persistence. A profound knowledge about
the intrinsic properties of HCV and its effects on intrahe-
patic immunity is essential to be able to design effective
immunotherapies against HCV such as therapeutic HCV
vaccines.
Keywords HCV Kupffer cells Dendritic cells
NK cells NKT cells Adaptive immune response
The Role of Glycyrrhizin, an Inhibitor of HMGB1 Protein,
in Anticancer Therapy
Ryszard Smolarczyk • Tomasz Cichon´ • Sybilla Matuszczak • Iwona Mitrus •
Marta Lesiak • Magdalena Kobusin´ ska • Wojciech Kamysz • Magdalena Jarosz •
Aleksander Sieron´ • Stanisław Szala
Received: 8 November 2011 / Accepted: 11 May 2012 / Published online: 25 August 2012
Ó L. Hirszfeld Institute of Immunology and Experimental Therapy, Wroclaw, Poland 2012
Abstract Certain anticancer drugs, such as the peptide
CAMEL (aa sequence KWKLFKKIGAULKVL) induce
necrotic type of cell death. During this process, a protein
termed high mobility group box 1 (HMGB1) is released
from cell nucleus into cytoplasm and then into extracellular
milieu. Outside of cells, it becomes a proinflammatory
cytokine. Its effects range from stimulation of cancer as
well as endothelial cell proliferation, to activation of
angiogenesis, cell motility and induction of inflammatory
conditions. Release of HMGB1 cytokine during the course
of anticancer therapy has negative effects upon the therapy
itself, since it leads to tumor relapse. We assumed that the
inhibition of HMGB1 activity may be conducive towards
better therapeutic results in case of drugs inducing necrotic
cell death. In this context we studied glycyrrhizin (GR), a
triterpenoid saponin glycoside of glycyrrhizic acid and a
well-known inhibitor of HMGB1. We have shown that GR
inhibits proliferation and migration of cells stimulated by
HMGB1 cytokine, as well as HMGB1-induced formation
of blood vessels and reduces inflammatory condition
(lowering tumor necrosis factor a levels). GR-mediated
inhibition of HMGB1 activity (CAMEL-induced release)
impedes, in turn, tumor regrowth in mice. As expected,
inhibited tumor regrowth is linked to diminished tumor
levels of the released HMGB1 and reduced inflammatory
condition. To conclude, the use of GR significantly
improved anticancer effectiveness of the CAMEL peptide.
Keywords Anticancer peptide Glycyrrhizin
HMGB1 Necrosis Tumor therapy
Retraction Note: Impact of Different Treatment of Whole-Body
Cryotherapy on Circulatory Parameters
Felice Giulio Bonomi • Massimo De Nardi •
Aldo Fappani • Viviana Zani • Giuseppe Banfi
Published online: 27 July 2012
Ó L. Hirszfeld Institute of Immunology and Experimental Therapy, Wroclaw, Poland 2012
Retraction to: Arch. Immunol. Ther. Exp. (2012)
60:145–150
DOI 10.1007/s00005-012-0163-4
The Editor-in-Chief of ‘‘Archivum Immunologiae et
Therapiae Experimentalis’’, in agreement with the authors
and the publisher, hereby retracts the article entitled
‘‘Impact of different treatment of whole-body cryotherapy
on circulatory parameters’’, by Bonomi FG, De Nardi M,
Fappani A, Zani V, Banfi G; published in ‘‘Archivum
Immunologiae et Therapiae Experimentalis’’ (2012),
60:145–150. This article has been retracted due to copy-
right issues that cannot be resolved
The Classification of Microglial Activation Phenotypes
on Neurodegeneration and Regeneration in Alzheimer’s
Disease Brain
Megan M. Varnum • Tsuneya Ikezu
Received: 2 November 2011 / Accepted: 20 February 2012 / Published online: 19 June 2012
Ó L. Hirszfeld Institute of Immunology and Experimental Therapy, Wroclaw, Poland 2012
Abstract Alzheimer’s disease (AD) is a neurodegen-
erative disease characterized by progressive decline of
cognitive function. There is no therapy that can halt or
reverse its progression. Contemporary research suggests
that age-dependent neuroinflammatory changes may play
a significant role in the decreased neurogenesis and
cognitive impairments in AD. The innate immune
response is characterized by pro-inflammatory (M1)
activation of macrophages and subsequent production of
specific cytokines, chemokines, and reactive intermedi-
ates, followed by resolution and alternative activation for
anti-inflammatory signaling (M2a) and wound healing
(M2c). We propose that microglial activation phenotypes
are analogous to those of macrophages and that their
activation plays a significant role in regulating neuro-
genesis in the brain. Microglia undergo a switch from an
M2- to an M1-skewed activation phenotype during
aging. This review will assess the neuroimmunological
studies that led to characterization of the different
microglial activation states in AD mouse models. It will
also discuss the roles of microglial activation on neuro-
genesis in AD and propose anti-inflammatory molecules
as exciting therapeutic targets for research. Molecules
such as interleukin-4 and CD200 have proven to be
important anti-inflammatory mediators in the regulation
of neuroinflammation in the brain, which will be dis-
cussed in detail for their therapeutic potential.
Keywords Alzheimer’s disease Microglia
Neurogenesis Neuroinflammation
Effects of the Histone Deacetylase Inhibitor, Trichostatin A,
in a Chronic Allergic Airways Disease Model in Mice
Simon G. Royce • William Dang • Gao Yuan • Jenny Tran •
Assam El-Osta • Tom C. Karagiannis • Mimi L. K. Tang
Received: 24 July 2011 / Accepted: 20 February 2012 / Published online: 9 June 2012
Ó L. Hirszfeld Institute of Immunology and Experimental Therapy, Wroclaw, Poland 2012
Abstract There is a need for new asthma therapies that
can concurrently address airway remodeling, airway
hyperresponsiveness and progressive irreversible loss of
lung function, in addition to inhibiting inflammation. His-
tone deacetylase inhibitors (HDACi) alter gene expression
by interfering with the removal of acetyl groups from
histones. The HDACi trichostatin A (TSA) has pleiotropic
effects targeting key pathological processes in asthma
including inflammation, proliferation, angiogenesis and
fibrosis. The aim was to evaluate the effects of TSA
treatment in a mouse model of chronic allergic airways
disease (AAD). Wild-type BALB/c mice with AAD were
treated intraperitoneally with 5 mg/kg TSA or vehicle
control. Airway inflammation was assessed by bronchoal-
veolar lavage fluid (BALF) cell counts and histological
examination of lung tissue sections. Remodeling was
assessed by morphometric analysis and airway hyperre-
sponsiveness was assessed by invasive plethysmography.
TSA-treated mice had a reduced number of total inflam-
matory cells and eosinophils within the BALF as compared
to vehicle-treated mice (both p \ 0.05). Furthermore,
airway remodeling changes were significantly reduced with
TSA compared to vehicle-treated mice, with fewer goblet
cells (p \ 0.05), less subepithelial collagen deposition
(p \ 0.05) and attenuated airway hyperresponsiveness at
the highest methacholine dose. These findings demonstrate
that treatment with an HDACi can concurrently reduce
structural airway remodeling changes and airway hyper-
responsiveness, in addition to attenuating airway
inflammation in a chronic AAD model. This has important
implications for the development of novel treatments for
severe asthma.
Keywords Airway hyperresponsiveness
Airway remodeling Asthma Trichostatin A Therapy
Biological Roles of Liver X Receptors in Immune Cells
Mo´nica Pascual-Garcı´a • Annabel F. Valledor
Received: 2 November 2011 / Accepted: 16 March 2012 / Published online: 14 June 2012
Ó L. Hirszfeld Institute of Immunology and Experimental Therapy, Wroclaw, Poland 2012
Abstract Liver X receptors (LXRs) are members of the
nuclear receptor superfamily that are activated by specific
oxysterols. LXRs heterodimerize with retinoid X receptors
to regulate positively the expression of a variety of target
genes, many of which are involved in lipid and glucose
metabolism. In the last few years, new targets of LXR
activation have been identified with roles in the modulation
of immune responses. Moreover, LXRs mediate repression
of inflammatory pathways through mechanisms collec-
tively known as transrepression. Here, we revise recent
findings on the impact of LXR activation on immune
responses, with an emphasis on advances in the under-
standing of the molecular mechanisms that mediate these
effects.
Keywords LXR Transrepression Inflammation
Gene expression
Bactericidal Effects and Mechanisms of Visible Light-Responsive
Titanium Dioxide Photocatalysts on Pathogenic Bacteria
Je-Wen Liou • Hsin-Hou Chang
Received: 29 August 2011 / Accepted: 28 February 2012 / Published online: 8 June 2012
Ó L. Hirszfeld Institute of Immunology and Experimental Therapy, Wroclaw, Poland 2012
Abstract This review focuses on the antibacterial activi-
ties of visible light-responsive titanium dioxide (TiO2)
photocatalysts. These photocatalysts have a range of
applications including disinfection, air and water cleaning,
deodorization, and pollution and environmental control.
Titanium dioxide is a chemically stable and inert material,
and can continuously exert antimicrobial effects when
illuminated. The energy source could be solar light; there-
fore, TiO2 photocatalysts are also useful in remote areas
where electricity is insufficient. However, because of its
large band gap for excitation, only biohazardous ultraviolet
(UV) light irradiation can excite TiO2, which limits its
application in the living environment. To extend its appli-
cation, impurity doping, through metal coating and
controlled calcination, has successfully modified the sub-
strates of TiO2 to expand its absorption wavelengths to the
visible light region. Previous studies have investigated the
antibacterial abilities of visible light-responsive photocat-
alysts using the model bacteria Escherichia coli and human
pathogens. The modified TiO2 photocatalysts significantly
reduced the numbers of surviving bacterial cells in response
to visible light illumination. They also significantly reduced
the activity of bacterial endospores; reducing their toxicity
while retaining their germinating abilities. It is suggested
that the photocatalytic killing mechanism initially damages
the surfaces weak points of the bacterial cells, before totally
breakage of the cell membranes. The internal bacterial
components then leak from the cells through the damaged
sites. Finally, the photocatalytic reaction oxidizes the cell
debris. In summary, visible light-responsive TiO2 photo-
catalysts are more convenient than the traditional UV light-
responsive TiO2 photocatalysts because they do not require
harmful UV light irradiation to function. These photocata-
lysts, thus, provide a promising and feasible approach for
disinfection of pathogenic bacteria; facilitating the pre-
vention of infectious diseases.
Keywords Photocatalysis Titania Bactericidal effect
Visible light responsive Bacterial spore Mode of action
Low Frequency of Regulatory T Cells in the Peripheral Blood
of Children with Type 1 Diabetes Diagnosed under the Age of Five
Agnieszka Szypowska • Anna Stelmaszczyk-Emmel •
Urszula Demkow • Włodzimierz Łuczyn´ ski
Received: 20 October 2011 / Accepted: 11 May 2012 / Published online: 9 June 2012
Ó L. Hirszfeld Institute of Immunology and Experimental Therapy, Wroclaw, Poland 2012
Abstract The highest annual increase in the incidence of
type 1 diabetes (T1D) in children under the age of 5 years
and aggressive process of b-cell destruction in this age
group indicate the need to assess the immune system. The
aim of this study was to evaluate regulatory T cells (Tregs)
frequency in the peripheral blood of children \5 years of
age with newly diagnosed T1D in comparison with diabetic
children diagnosed at a later age and healthy controls. 40
children with newly diagnosed T1D (20 children \5 years
of age and 20 older patients) and 40 age-matched controls
were included in this study. Flow cytometric analysis of
Tregs was performed using the following markers: CD4,
CD25, CD127, FoxP3, IL-10, and TGF-b. Apoptosis was
measured using anti-active caspase 3 monoclonal antibody.
Fasting C-peptide and HbA1c were monitored as well. We
showed that T1D children \5 years had lower C-peptide
concentration than diabetic children C5 years of age (0.32
vs. 0.80 ng/ml, respectively, p = 0.0005). There was lower
frequency of CD4?CD25highCD127lowFoxP3? Tregs in
T1D children \5 years than C5 years of age (0.87 vs.
1.56 %, respectively, p = 0.017). Diabetic children
\5 years had lower CD4?CD25highCD127lowFoxP3?,
CD4?CD25highIL-10, and CD4?CD25highTGF-b Tregs
compared to age-matched controls. There was no differ-
ence in Tregs apoptosis between the examined groups. This
study highlights the distinctiveness of diabetes in children
\5 years of age. Understanding the differences of immune
system activity in the young diabetic children would open
the way to identify children at risk for T1D and enables the
use of novel forms of intervention.
Keywords Tregs FoxP3 IL-10 TGF-b
The NADPH Oxidase Family and its Inhibitors
Paulina Kleniewska • Aleksandra Piechota •
Beata Skibska • Anna Gora˛ca
Received: 23 September 2011 / Accepted: 27 March 2012 / Published online: 14 June 2012
Ó L. Hirszfeld Institute of Immunology and Experimental Therapy, Wroclaw, Poland 2012
Abstract The classical nicotinamide adenine dinucleo-
tide phosphate (NADPH) oxidase was originally detected
in neutrophils as a multicomponent enzyme that catalyzes
the generation of superoxide from oxygen and the reduced
form of NADPH. This enzyme is composed of two mem-
brane-bound subunits (p22phox and gp91phox), three
cytosolic subunits (p67phox, p47phox, and p40phox) and a
small G-protein Rac (Rac1 and Rac2). Recently, it has been
demonstrated that there are several isoforms of nonphag-
ocytic NADPH oxidase. Endothelial cells, vascular smooth
muscle cells or adventitial fibroblasts possess multiple
isoforms of this enzyme. The new homologs, along with
gp91phox are now designated the Nox family of NADPH
oxidases and are key sources of reactive oxygen species in
the vasculature. Reactive oxygen species play a signifi-
cant role in regulating endothelial function and vascular
tone. However, besides the participation in the processes
of physiological cell, these enzymes can also be the per-
petrator of oxidative stress that causes endothelial
dysfunction. This review summarizes the current state of
knowledge of the structure and functions of NADPH
oxidase and NADPH oxidase inhibitors in the treatment of
disorders with endothelial damage.
Keywords NADPH oxidase Nox family
Nox inhibitors Reactive oxygen species
Cardiovascular disease
Optimizing the European Regulatory Framework for Sustainable
Bacteriophage Therapy in Human Medicine
Gilbert Verbeken • Jean-Paul Pirnay •
Daniel De Vos • Serge Jennes • Martin Zizi •
Rob Lavigne • Minne Casteels • Isabelle Huys
Received: 3 January 2012 / Accepted: 21 February 2012 / Published online: 17 April 2012
Ó L. Hirszfeld Institute of Immunology and Experimental Therapy, Wroclaw, Poland 2012
Abstract For practitioners at hospitals seeking to use
natural (not genetically modified, as appearing in nature)
bacteriophages for treatment of antibiotic-resistant bacte-
rial infections (bacteriophage therapy), Europe’s current
regulatory framework for medicinal products hinders more
than it facilitates. Although many experts consider bacte-
riophage therapy to be a promising complementary (or
alternative) treatment to antibiotic therapy, no bacterio-
phage-specific framework for documentation exists to date.
Decades worth of historical clinical data on bacteriophage
therapy (from Eastern Europe, particularly Poland, and the
former Soviet republics, particularly Georgia and Russia,
as well as from today’s 27 EU member states and the US)
have not been taken into account by European regulators
because these data have not been validated under current
Western regulatory standards. Consequently, applicants
carrying out standard clinical trials on bacteriophages in
Europe are obliged to initiate clinical work from scratch.
This paper argues for a reduced documentation threshold
for Phase 1 clinical trials of bacteriophages and maintains
that bacteriophages should not be categorized as classical
medicinal products for at least two reasons: (1) such a
categorization is scientifically inappropriate for this spe-
cific therapy and (2) such a categorization limits the
marketing authorization process to industry, the only
stakeholder with sufficient financial resources to prepare a
complete dossier for the competent authorities. This paper
reflects on the current regulatory framework for medicines
in Europe and assesses possible regulatory pathways for the
(re-)introduction of bacteriophage therapy in a way that
maintains its effectiveness and safety as well as its inherent
characteristics of sustainability and in situ self-amplifica-
tion and limitation.
Keywords Bacteriophage Therapy Human
European Regulatory Legal
Matched Sibling Versus Matched Unrelated Allogeneic
Hematopoietic Stem Cell Transplantation in Children with Severe
Acquired Aplastic Anemia: Experience of the Polish Pediatric
Group for Hematopoietic Stem Cell Transplantation
Dawid Szpecht • Ewa Gorczyn´ ska • Krzysztof Kałwak • Joanna Owoc-Lempach • Marta Choma •
Jan Styczyn´ ski • Jolanta Goz´dzik • Agnieszka Dłu_zniewska • Mariusz Wysocki • Jerzy R. Kowalczyk •
Alicja Chybicka • Anna Pieczonka • Jacek Wachowiak
Received: 1 April 2011 / Accepted: 10 February 2012 / Published online: 17 April 2012
Ó L. Hirszfeld Institute of Immunology and Experimental Therapy, Wroclaw, Poland 2012
Abstract In the study, 48 children with severe acquired
aplastic anemia (SAA) transplanted from matched sibling
donor (MSD) between 1991 and 2009, and 38 children with
SAA transplanted from matched unrelated donor (MUD)
between 2000 and 2009 were evaluated. Engraftment was
achieved in 45 (93.75 %) patients after MSD-hematopoi-
etic stem cell transplantation (HSCT) and in 33 (86.8 %)
after MUD-HSCT. Transplant-related mortality rate after
MSD-HSCT was 8 %, while 37 % after MUD-HSCT.
After MSD-HSCT 44 (91.7 %) patients are alive for
1–216 months (median: 85 months), while after MUD-
HSCT 24 (63.2 %) patients for 1–84 months (median:
16 months). The 5-year probability of event-free survival
after MSD-HSCT and MUD-HSCT was 87 and 53 %,
respectively, while 5 years of overall survival was 91 and
64 %, respectively. It was concluded that MSD-HSCT as
the first line treatment for children with SAA is a safe
therapeutic approach with a low rate of treatment failures
and excellent outcome. Results of MUD-HSCT in pediatric
patients with SAA who failed to respond to immunosup-
pressive therapy are still inferior than those of MSD-
HSCT. Treatment failures of MUD-HSCT are mainly
related to infectious complications and graft failure. It
seems, however, that HLA-matching of unrelated donors at
allelic level along with early MUD-HSCT after FCA
(FLUDA, low-dose cyclophosphamide, and anti-thymocyte
globulin) conditioning, perhaps using lower Thymoglobu-
lin dose could enable further improvement of long-term
results in children with SAA who lack MSD.
Keywords Severe acquired aplastic anemia Children
Allogeneic hematopoietic stem cell transplantation
Matched sibling donor Matched unrelated donor
Myasthenia, Spondylitis and Enterococcus faecalis Endocarditis
Claudia Sto¨llberger • Markus Thalmann •
Josef Finsterer
Received: 15 July 2011 / Accepted: 13 December 2011 / Published online: 10 April 2012
Ó L. Hirszfeld Institute of Immunology and Experimental Therapy, Wroclaw, Poland 2012
Abstract Parasympathomimetics, immunosuppression
and plasmapheresis have considerably improved manage-
ment and prognosis of myasthenia gravis. Side effects of
these measures, however, may complicate the course of the
disease. In a 66-year-old male with myasthenia gravis and
lower back pain, blood cultures, echocardiography and
magnetic resonance imaging led to the diagnosis of endo-
carditis and spondylodiscitis. Enterococcus faecalis grew
in the blood cultures as well as on the aortic and tricuspid
valve vegetations which were resected during cardiac sur-
gery. Possible sources of the infection might be E. faecalis
infections of catheter tips during a 46-day stay in the
intensive care unit 11 months earlier where he had under-
gone plasmapheresis, hemodiafiltration and mechanical
ventilation, or recurrent diarrheas since 18 months.
Infection was favored by immunosuppression with gluco-
corticoids and azathioprine which received the patient
because of myasthenia gravis and hypothyroidism. Patients
with myasthenia gravis require close follow up, including
infection parameters, especially when they receive immu-
nosuppressive therapy and when microorganisms known to
cause endocarditis, are identified.
Keywords Endocarditis Enterococcus faecalis
Myasthenia gravis
Therapeutic Potential of Induced and Natural FoxP3+ Regulatory
T Cells for the Treatment of Graft-Versus-Host Disease
Immo Prinz • Christian Koenecke
Received: 10 October 2011 / Accepted: 4 January 2012 / Published online: 5 April 2012
Ó L. Hirszfeld Institute of Immunology and Experimental Therapy, Wroclaw, Poland 2012
Abstract Graft-versus-host disease (GvHD) remains a
major complication after allogeneic hematopoietic stem-
cell-transplantation. Present GvHD prophylaxis and treat-
ment is still based on unspecific immunosuppressive drug
therapy. Over the last decade, the potential of cell-based
therapies involving the infusion of regulatory T cells has
emerged as a feasible alternative approach for the treatment
and prevention of GvHD. Here we review current efforts to
translate data obtained in rodent models into clinical trials.
Special emphasis is placed on the variety of strategies to
generate sufficient numbers of alloantigen-specific regula-
tory T cells for adoptive cell therapy. This can be achieved
either by expansion or by induction of a regulatory phe-
notype in naive T cells. Stability of the immunosuppressive
phenotype of transferred regulatory T cells even in the
highly inflammatory environment of acute GvHD will be
thereby a critical parameter for actual therapeutic
application.
Keywords GvHD Bone marrow transplantation
FoxP3? regulatory T cells
The Role and Regulation of mTOR in T-Lymphocyte Function
Thomas F. O’Brien • Xiao-Ping Zhong
Received: 12 November 2011 / Accepted: 30 January 2012 / Published online: 8 April 2012
Ó L. Hirszfeld Institute of Immunology and Experimental Therapy, Wroclaw, Poland 2012
Abstract The conversion of naı¨ve T cells into effector T
cells is initiated by stimulation through the T-cell receptor
(TCR). Upon activation, T cells undergo significant mor-
phological and functional changes, putting new metabolic
demands on the cell. Past research has identified the
mammalian target of rapamycin (mTOR) as a critical
regulator of cell metabolism, and the development of new
genetic models has begun to reveal an important role for
this pathway in the homeostasis and function of T lym-
phocytes. In this review, we focus on the most recent
findings that demonstrate the ability of mTOR to regulate
T-cell activation, CD8? memory cell formation and func-
tion, and helper T lineage differentiation. Furthermore, we
highlight the importance of tight control of mTOR sig-
naling by tuberous sclerosis complex 1 for T-cell
homeostasis, and the regulation of mTOR signaling by
diacylglycerol kinases and the RasGRP1-Ras-Erk1/2
pathway in the context of TCR signaling.
Keywords Mammalian target of rapamycin
Tuberous sclerosis complex 1 Diacylglycerol kinase
RasGPP1 PI3K/Akt T lymphocyte
Lactobacillus rhamnosus Exopolysaccharide Ameliorates
Arthritis Induced by the Systemic Injection of Collagen
and Lipopolysaccharide in DBA/1 Mice
Bernadeta Nowak • Marta Ciszek-Lenda •
Małgorzata S´ ro´ttek • Andrzej Gamian • Ewa Kontny •
Sabina Go´rska-Fra˛czek • Janusz Marcinkiewicz
Received: 3 June 2011 / Accepted: 12 December 2011 / Published online: 8 April 2012
Ó L. Hirszfeld Institute of Immunology and Experimental Therapy, Wroclaw, Poland 2012
Abstract Oral administration of some probiotic bacteria
(e.g. Lactobacillus rhamnosus) attenuates various types of
experimental arthritis, including collagen-induced arthritis
(CIA) and inhibits arthritogenic autoantibodies. Much less
is known about the possible anti-arthritogenic properties of
exopolysaccharide (EPS), the major component of lactic
bacteria biofilm. In this study, we asked the question
whether systemic administration of EPS derived from
L. rhamnosus KL37 depresses the production of anti-collagen
IgG and affects the development of CIA in DBA/1 mice.
Arthritis was induced employing two models of active
CIA, in which mice were immunized with type II collagen
(CII) either in the presence of lipopolysaccharide (LPS;
mild arthritis with moderate CII-specific IgG production)
or with Complete Freund’s Adjuvant and LPS (severe
arthritis with massive CII-specific IgG production). Passive
CIA was induced by intravenous injection of CII-specific
monoclonal antibodies and LPS. Disease progression, the
incidence and severity of arthritis, were determined. Serum
concentration of CII-specific IgG was measured by
enzyme-linked immunosorbent assay. Systemic adminis-
tration of EPS markedly reduced CII-specific antibody
production. Moreover, EPS significantly ameliorated
arthritis in the active models of CIA, especially, when LPS
alone was used as an adjuvant. In contrast, when arthrito-
genic antibodies were injected to mice in high amounts, the
effect of EPS on the development of passive CIA was
negligible and transient. These results show that EPS can
suppress active CIA by the inhibition of arthritogenic
antibodies production. Therefore, we suggest that EPS or
EPS-producing probiotics may be promising agents for the
supporting therapy of patients with rheumatoid arthritis.
Keywords Exopolysaccharide Lactobacillus
rhamnosus Collagen-induced arthritis LPS
Collagen-specific antibodies
Prostate Cancer and Immunoproteome: Awakening
and Reprogramming the Guardian Angels
Ammad Ahmad Farooqi • Sundas Fayyaz •
Muhammad Zahid Qureshi • Sadia Rashid
Received: 7 September 2011 / Accepted: 7 March 2012 / Published online: 8 April 2012
Ó L. Hirszfeld Institute of Immunology and Experimental Therapy, Wroclaw, Poland 2012
Abstract Prostate cancer is a life-threatening molecular
disorder that is undruggable to date because of stumbling
blocks in the standardization of therapy. An emerging
framework of research is addressing how pathways that are
derailed during tumorigenesis are linked to immunological
responses, which are instrumental in immunosurveillance
of cancer. However, interestingly, cancer cells circumvent
such immunosurveillance through development of poorly
immunogenic tumor cell variants (immunoselection) and
through subversion of the immunological nanomachinery
(immunosubversion). Detailed mechanistic insights of
molecular specificities that regulate natural killer (NK) cell
function suggest that it might be promising to design NK
cell-based immunotherapeutic interventions against pros-
tate cancer. Here, we elucidate evidence for NK cell
targeting of prostate cancer proteome and address critical
questions that, in our view, need thoughtfulness for the
development of successful NK cell-based therapies. This
review also disproves our contemporary understanding of
the versatile regulators of DNA damage repair (ATM,
ATR) that trigger cell surface expression of NKG2D
ligands and consequent elimination of the tumor cells by
NK cells and other lymphocytes that express NK cell
receptors. Substantial fraction of information has been
generated that guarantees productive future for this tech-
nology as more optimized constructs, better trial designs,
and improved platforms are being brought from benchtop
to bedside.
Keywords Prostate cancer Immunoproteome
DNA damage Major histocompatibility complex proteins
Bacterial Cell Surface Structures in Yersinia enterocolitica
Nataniel Białas • Katarzyna Kasperkiewicz •
Joanna Radziejewska-Lebrecht • Mikael Skurnik
Received: 1 July 2011 / Accepted: 30 January 2012 / Published online: 8 April 2012
Ó L. Hirszfeld Institute of Immunology and Experimental Therapy, Wroclaw, Poland 2012
Abstract Yersinia enterocolitica is a widespread member
of the family of Enterobacteriaceae that contains both non-
virulent and virulent isolates. Pathogenic Y. enterocolitica
strains, especially belonging to serotypes O:3, O:5,27, O:8
and O:9 are etiologic agents of yersiniosis in animals and
humans. Y. enterocolitica cell surface structures that play a
significant role in virulence have been subject to many
investigations. These include outer membrane (OM) glyco-
lipids such as lipopolysaccharide (LPS) and enterobacterial
common antigen (ECA) and several cell surface adhesion
proteins present only in virulent Y. enterocolitica, i.e., Inv,
YadA and Ail. While the yadA gene is located on the Yersinia
virulence plasmid the Ail, Inv, LPS and ECA are chromo-
somally encoded. These structures ensure the correct
architecture of the OM, provide adhesive properties as well
as resistance to antimicrobial peptides and to host innate
immune response mechanisms.
Keywords Yersinia enterocolitica Lipopolysaccharide
Enterobacterial common antigen
Yersinia virulence plasmid
Talin, Vinculin and Nestin Expression in Orofacial Muscles
of Dystrophin Deficient mdx Mice
Alexander Spassov • Tomasz Gredes • Dragan Pavlovic •
Tomasz Gedrange • Christian Lehmann • Silke Lucke •
Christiane Kunert-Keil
Received: 8 February 2011 / Accepted: 26 September 2011 / Published online: 4 February 2012
Ó L. Hirszfeld Institute of Immunology and Experimental Therapy, Wroclaw, Poland 2012
Abstract The activity of cytoskeletal proteins like talin,
vinculin and nestin increases in muscle that regenerates.
Little is known about their role or at least their expression
in the process of regeneration in masticatory muscles of
mdx mice, a model of Duchenne muscular dystrophy. To
determine a potential role of cytoskeletal proteins in the
regeneration process of mdx masticatory muscles, we
examined the expression of talin 1, talin 2, vinculin and
nestin in 100-day-old control and mdx mice using quanti-
tative RT-PCR, Western blot analyses and histochemistry.
The protein expression of talin 1, talin 2, nestin and vin-
culin in mdx muscles remained unchanged as compared
with normal mice. However, in mdx masseter it was found
a relative increase of nestin compared to controls. The
protein expression of talin 1 and vinculin tended to be
increased in mdx tongue and talin 2 to diminish in mdx
masseter and temporal muscle. In mdx mice, we found
significantly lower percentage of transcripts coding for
nestin, talin 1, talin 2 and vinculin in masseter (p \ 0.05)
and temporal muscle (p \ 0.001). In contrast, the mRNA
expression of nestin was found to be increased in mdx
tongue. Activated satellite cells, myoblasts and immature
regenerated muscle fibres in mdx masseter and temporal
revealed positive staining for nestin. The findings of the
presented work suggest dystrophin-lack-associated changes
in the expression of cytoskeletal proteins in mdx mastica-
tory muscles could be compensatory for dystrophin
absence. The expression of nestin may serve as an indicator
for the regeneration in the orofacial muscles.
Keywords mdx Nestin Regeneration Dystrophy
Masticatory mucles
Sepsis Immunopathology: Perspectives of Monitoring
and Modulation of the Immune Disturbances
Tomasz Skirecki • Urszula Borkowska-Zielin´ska •
Małgorzata Złotorowicz • Gra_zyna Hoser
Received: 4 July 2011 / Accepted: 7 November 2011 / Published online: 3 February 2012
Ó L. Hirszfeld Institute of Immunology and Experimental Therapy, Wroclaw, Poland 2012
Abstract Septic syndromes are the main cause of death in
the intensive care units and although the mortality rates is
slowly decreasing, the occurrence of the disease has been
increasing. The pathogenesis of sepsis includes countless
disturbances of the host immune system starting with a
harmful, infection-triggered exaggerated inflammatory
cascade, followed by the development of an immunopa-
ralysis state. The latter contributes to the failure in pathogen
eradication and leads to secondary infections, which are
often the cause of fatal complications. In this review, we
consider different novel therapeutic strategies for restora-
tion of immune function. The use of glucocorticoids,
intravenous immunoglobulins, heparin, recombinant human
activated protein C, granulocyte macrophage colony-stim-
ulating factor, granulocyte colony-stimulating factor,
interferon-c, statins, macrolides and high-volume hemofil-
tration are discussed. Even though some clinical trials of
these regimens are promising, the key to their successful
application seems to be the precise monitoring of the status
of immune system followed by implementation of the
adequate therapy. Thus, in this paper we present distur-
bances in the immune system in the course of human sepsis,
with special attention to the parameters that could be
monitored and serve as markers for immunomodulatory
therapies. We conclude by briefly presenting the current
sepsis treatment strategy.
Keywords Sepsis Monitoring Flow cytometry
Immunomodulation
Microvesicles in Health and Disease
Jameel M. Inal • Ephraim A. Ansa-Addo •
Dan Stratton • Sharad Kholia • Samuel S. Antwi-Baffour •
Samireh Jorfi • Sigrun Lange
Received: 14 July 2011 / Accepted: 10 November 2011 / Published online: 4 February 2012
Ó L. Hirszfeld Institute of Immunology and Experimental Therapy, Wroclaw, Poland 2012
Abstract Microvesicles (or MVs) are plasma membrane-
derived vesicles released from most eukaryotic cells con-
stitutively during early apoptosis or at higher levels after
chemical or physical stress conditions. This review looks at
some of the functions of MVs in terms of intercellular
communication and ensuant signal transduction, including
the transport of proteins (unconventional protein export) as
well as of mRNA and microRNA. MVs also have roles in
membrane repair, the removal of misfolded proteins, and in
the control of apoptosis. We also discuss the role MVs have
been shown to have in invasive growth and metastasis as
well as in hypoxia in tumours and cerebral ischaemia. The
association of MVs in infectious and autoimmune disease
is also summarised together with their possible use as
therapeutic agents.
Keywords Microvesicles Intercellular transport
Cancer Autoimmune disease Stroke Infectious disease
Antibodies in the Treatment of Aplastic Anemia
David Go´mez-Almaguer • Jose Carlos Jaime-Pe´rez •
Guillermo J. Ruiz-Arguelles
Received: 1 July 2011 / Accepted: 28 November 2011 / Published online: 4 February 2012
Ó L. Hirszfeld Institute of Immunology and Experimental Therapy, Wroclaw, Poland 2012
Abstract Antibodies have been the cornerstone of
treatment of acquired aplastic anemia for more than
25 years. Treatment with antithymocyte globulin (ATG)
is considered pivotal and the addition of cyclosporine
improves the overall response rate. This antibody is het-
erogeneous and horse ATG is apparently more effective
than rabbit ATG. Several issues remain unsolved in
relation to the combination of ATG and cyclosporine:
cost, toxicity and late clonal disorders. In recent years,
alternative immunosuppressive therapy has been proposed
and new antibodies have emerged: porcine ATG, ale-
mtuzumab, daclizumab, and rituximab. Experience with
these antibodies is limited to a few studies with ale-
mtuzumab being the most promising, but the results are
interesting and provocative. More studies are needed to
find the perfect antibody.
Keywords Aplastic anemia Antibodies
Alemtuzumab Antithymocyte globulin
Impact of Different Treatment of Whole-Body Cryotherapy
on Circulatory Parameters
Felice Giulio Bonomi • Massimo De Nardi •
Aldo Fappani • Viviana Zani • Giuseppe Banfi
Alpha-1 Antitrypsin: A Potent Anti-Inflammatory
and Potential Novel Therapeutic Agent
David A. Bergin • Killian Hurley • Noel G. McElvaney •
Emer P. Reeves
Received: 15 June 2011 / Accepted: 23 November 2011 / Published online: 15 February 2012
Ó L. Hirszfeld Institute of Immunology and Experimental Therapy, Wroclaw, Poland 2012
Abstract Alpha-1 antitrypsin (AAT) has long been thought
of as an important anti-protease in the lung where it is known
to decrease the destructive effects of major proteases such as
neutrophil elastase. In recent years, the perception of this
protein in this simple one dimensional capacity as an anti-
protease has evolved and it is now recognised that AAT has
significant anti-inflammatory properties affecting a wide
range of inflammatory cells, leading to its potential thera-
peutic use in a number of important diseases. This present
review aims to discuss the described anti-inflammatory
actions of AAT in modulating key immune cell functions,
delineate known signalling pathways and specifically to
identify the models of disease in which AAT has been shown
to be effective as a therapy.
Keywords Alpha-1 antitrypsin Innate immune cells
Alpha-1 antitrypsin deficiency Replacement therapy
Immunotherapy of Patients with Recurrent Spontaneous
Miscarriage and Idiopathic Infertility: Does the
Immunization-Dependent Th2 Cytokine
Overbalance Really Matter?
Jacek R. Wilczyn´ ski • Paweł Radwan •
Henryk Tcho´rzewski • Małgorzata Banasik
Received: 17 March 2011 / Accepted: 11 July 2011 / Published online: 18 February 2012
Ó L. Hirszfeld Institute of Immunology and Experimental Therapy, Wroclaw, Poland 2012
Abstract Recurrent spontaneous miscarriage (RSM) and
idiopathic infertility (IIF) are partially caused by immuno-
logic disturbances. Paternal lymphocyte immunization (PLI)
is proposed for restoration of the proper Th1/Th2 balance in
these patients, but still there are controversies on PLI
mechanism, its efficacy and identification of patients who
may benefit from this therapy. The study group consisted of
n = 34 RSM and n = 42 IIF women with unexplained
miscarriage or IIF. PLI was offered as a treatment in both
groups. Peripheral blood lymphocyte (PBL) populations
(CD3?, CD3–/CD19?, CD3?/CD4?, CD3?/CD8?, CD3–/
CD16?CD56?) were studied before immunization, while
PBL cytokine secretion (IFN-c, TNF-a, IL-10, IL-5, IL-4,
IL-2), before and after immunization, pre-conceptionally in
both groups. The reference PBL ratio and cytokine levels
were adopted from previously studied normal fertile women.
PBL populations, concentration and ratio of Th1/Th2 cyto-
kines did not differ between RSM and IIF patients.
Compared to the results observed in normal fertile women
the levels of IFN-c, TNF-a and IL-2 were higher, while IL-10
lower in both RSM and IIF patients (p \ 0.01). After
immunization a decrease of IFN-c (RSM and IIF groups) and
IL-4 and IL-10 (RSM group) were observed, as well as an
increase in TNF-a/IL-4 ratio (RSM group) (p \ 0.05). No
differences in Th1/Th2 concentration and ratio between
patients with successful and unsuccessful pregnancy were
observed. No significant correlations between success and
particular cytokine concentration were observed. Concen-
trations of Th1/Th2 cytokines and PBL populations did not
differ between RSM and IIF women. Th1 shift in both RSM
and IIF patients was observed in comparison to fertile
women. Treatment with PLI-induced pre-conceptionally
cytokine changes which neither indicated Th2 shift nor
correlated with subsequent pregnancy success.
Keywords Recurrent spontaneous miscarriage
Idiopathic infertility Cytokines
Paternal lymphocyte immunization