Treating autoimmune diseases through restoration of antigen-specific immune tolerance
Lawrence A. Wolfraim
Abstract
The first line of treatment for many human autoimmune diseases involves the use of anti-inflammatory or immunosuppressive drugs such as prednisone or other steroids that not only suppress the underlying autoimmune disease, but lead to global suppression of the immune system. The sequelae of this approach include increased risk of infection, carcinogenesis, and osteoporosis. Moreover, such broad spectrum immunosuppression tends to have transient therapeutic benefit, as in many cases the disease becomes refractory to these drugs. There is a clear need for more specific means to restore immune tolerance to the specific autoantigens implicated in disease pathology. This review provides an overview of some of these newer, more specific therapeutic approaches to restoring immune tolerance to autoantigens, with an emphasis on those approaches that have been or will soon be tested in controlled clinical trials. Covered here are peptide- or protein-based therapeutics, oral tolerance, and cellular and gene therapy approaches to restoring antigen-specific immune tolerance.
Key words: autoimmunity, immune tolerance, antigen-specific, T cell activation, clinical trial.
Regulation of innate and adaptive immune responses by the related cytokines IL-12, IL-23, and IL-27
Carol Beadling and Mark K. Slifka
Abstract
The functional characterization and subsequent purification of T cell growth factor/interleukin (IL)-2 in the early 1980s established this secreted protein as a key mediator of immune cell activation and provided the prototype that enabled the discovery of numerous cytokines over the ensuing two decades. While soluble immunoregulatory factors were initially identified functionally as biological activities present in the culture supernatants of activated lymphocytes/monocytes, this methodology shifted radically following the completion of the human genome sequence. Computer-generated structural modeling algorithms have replaced functional assays and biochemical purification as the initial means of discovering new cytokines. To date, a total of 31 interleukins, as well as over a dozen other related hematopoietic factors, have been identified. These cytokines and their receptors may be grouped on the basis of structural homologies as well as by shared ligand and receptor subunits. The challenge now at hand is to define the biological functions of the newly identified cytokines and to elucidate the common and divergent roles of related family members. This point is well illustrated by the IL-12/IL-23/IL-27 family, whose members share ligand and receptor subunits and play somewhat overlapping roles in innate and adaptive immune responses. These three cytokines are not entirely redundant, as they may preferentially activate naïve or memory T cells, induce discrete T cell cytokine profiles, contribute to distinct stages of host immune responses to infectious agents, and differentially promote autoimmunity. Further elucidation of the unique functions of the IL-12 family members may lead to improved immunodiagnostics and therapies.
Key words: immune regulation, T cell activation, cytokines.
Activation of
Salmonella-specific immune responses
in the intestinal mucosa
Aparna Srinivasan and Stephen J. McSorley
Department of Medicine, University of Connecticut Health Center, Farmington, Connecticut 06030, USA
Received: 2005.05.10, Accepted: 2005.07.11, Published online first: 2006.01.23
Summary
The mammalian immune response to Salmonella has long been a subject of scientific study. Indeed, many of the general
aspects of bacterial pathogenesis and host immune defense have been well described. However, a lack of clarity remains con-
cerning important aspects of the host immune response to Salmonella, particularly with regard to the induction of an immune
response in the intestinal mucosa. A major limitation has been the general lack of knowledge about specific antigenic targets
that are recognized by both the innate and adaptive immune response in the intestine. Progress towards the identification of
these targets is critical for the development of a detailed model of immunity to Salmonella and will lead to a better under-
standing of mucosal immune responses to other intracellular pathogens.
Key words: Salmonella, intestinal mucosa, immune responses
Stem cells in nephrology: present status and future
Ewa Wątorek and Marian Klinger
Department of Nephrology and Transplantation Medicine, Wroc³aw Medical University, Poland
Received: 2005.02.24, Accepted: 2005.07.18, Published online first: 2006.02.13
Abstract
Stem cell biology is currently developing rapidly because of the potential therapeutic utility of stem cells. The ability to
acquire any desired phenotype raises hope for regenerative therapies. Manipulation of these cells is a potentially valuable
tool; however, the mechanisms of stem cell differentiation and plasticity are currently beyond our control. In the field of
nephrology, the presence of adult kidney stem cells has been debated. Renal adult stem cells may be descendants of some
early kidney progenitors, or may be derived from bone marrow. Evidence of a hematopoietic stem-cell contribution to renal
repair encourages the possibility of bone marrow or stem cell transplantation as a means of treating autoimmune glomeru-
lopathies. The transplantation of fetal kidney tissue containing renal progenitors, which then develop into functional
nephrons, is a step towards renal regeneration. According to recent reports, the development of functional nephrons from
human mesenchymal stem cells in rodent whole-embryo culture is possible. Establishing in vitro self organs from autologous
stem cells would be a promising therapeutic solution in light of the shortage of allogenic organs and the unresolved problem
of chronic allograft rejection.
Key words: stem cells, embryonic stem cells, hematopoietic stem cells, metanephric mesenchyme, regenerative medicine,
stem cell diseases.
Therapeutic effects of cisplatin
on rat experimental autoimmune encephalomyelitis
Xiao−Bo Li1, 2 and Hermann J. Schluesener2
1 Institute of Environmental Medicine, Tongji Medical College, Huazhong Science and Technology University,
Wuhan City, China
2 Institute of Brain Research, University of Tuebingen, Tuebingen, Germany
Received: 2005.02.23, Accepted: 2005.08.10, Published online first: 2006.02.13
Abstract
Introduction: Experimental autoimmune encephalomyelitis (EAE) is a prototypic Th1-mediated autoimmune inflammatory
disease of the central nervous system (CNS), and serves as a model for the human demyelinating disease, multiple sclerosis.
Cisplatin is a drug widely used in the treatment of a variety of human neoplasias, such as advanced bladder carcinoma, adren-
al cortex carcinoma, breast cancer, head and neck or lung carcinoma. Cisplatin binds to DNA and interferes with cellular
repair and other mechanism, which eventually result into cell death. It is known that cisplatin can induce immunosuppres-
sive effects through inhibition of T cell activity. Therefore we analyzed the anti-inflammatory effects of cisplatin in a rat EAE
model.
Materials and Methods: EAE was induced in male LEW rats by immunizing with a synthetic peptide of guinea pig myelin basic
protein. The development of EAE and neurological signs were evaluated by a standard protocol. Immunohistochemistry was
applied to show immune cell infiltration into the CNS.
Results: Early treatment of EAE rats with cisplatin effectively ameliorated the development of disease and provided a sig-
nificant protective effect compared to control rats. Further, histological analysis demonstrated that the formation of the typ-
ical perivascular cuffs and brain infiltration of monocytes and lymphocytes were complete absent in cisplatin treated rats,
while abundant T cell infiltration was seen in the CNS of EAE rats.
Conclusions: Our data show that cisplatin has protective effects in EAE, indicating that cisplatin could be a candidate in the
treatment of human CNS autoimmunity.
Key words: experimental allergic encephalomyelitis, rat, cisplatin, suppression
Routine clinical laboratory tests correspond
to increased serum levels of 3-hydroxy fatty acids,
markers of endotoxins, in cardiosurgery patients
Leonard Kraśnik1, Bogumiła Szponar2, 4, Maciej Walczak3, Lennart Larsson4
and Andrzej Gamian2
1 Department of Physiology, Medical Academy of Poznañ, Poland
2 Institute of Immunology and Experimental Therapy, Polish Academy of Sciences, Wroc³aw, Poland
3 Cardiosurgery Clinical Hospital, Medical Academy of Poznañ, Poland
4 Department of Laboratory Medicine, Division of Medical Microbiology, Lund University, Sweden
Received: 2005.01.19, Accepted: 2005.08.29, Published online first: 2006.02.13
Abstract
Introduction: Endotoxemia developing during cardiosurgery as elevated endotoxin concentrations in patient’s serum may pre-
vail over 24 h after operation. A major reason is thought to be increased gut permeability resulting in endotoxin and bacte-
rial leakage. In this study we aimed to measure endotoxin levels on samples obtained during and after cardiovascular proce-
dures and compare them with clinical observations and laboratory test results.
Materials and Methods: 3-Hydroxy fatty acids (3-OH FAs) of 10–18 carbon chain length, chemical markers of endotoxin
(lipopolysaccharide), were determined in patient sera by gas chromatography-mass spectrometry-based analysis. Results
were compared with routine laboratory tests: blood morphology, urine, ALT, AST, bilirubin, kidney parameters, clotting
parameters, and gasometry.
Results: Of a total of 16 patients, 5 patients (group I) showed increased serum 3-OH FA levels and 11 patients (group II) did
not show any change in 3-OH FA levels 24 h after operation. All group I patients revealed leukocytosis, two developed post-
-operative anemia. Significantly different changes were observed: the initial, pre-operative 3-OH FA levels were similar for
both groups, while group I patients showed increased levels of all the studied 3-OH FAs during the operation (p≤0.05), and
3-OH C14 and 3-OH C16 remained elevated 24 h after the operation.
Conclusions: Cardiosurgery may strongly promote gut endotoxin translocation to the blood in some patients. Prolonged
leukocytosis, deep anemia, and increased liver dysfunction markers may indicate the need for observation of possible endo-
toxemia development. It is recommended to monitor the endotoxin level and/or endotoxemia markers in cardiosurgery
patients.
Key words: cardiosurgery, endotoxin, chemical markers, 3-hydroxy fatty acids.
Susceptibility of
Propionibacterium acnes
and Staphylococcus epidermidis to killing
by MPO-halide system products.
Implication for taurine bromamine as a new candidate
for topical therapy in treating acne vulgaris
Janusz Marcinkiewicz1, Rafał Biedroń1, Anna Białecka2, Andrzej Kasprowicz2,
Monika Mak1 and Marta Targosz3
1 Department of Immunology, Jagiellonian University Medical College, Cracow, Poland
2 Center of Microbiological Research and Autovaccines Ltd., Cracow, Poland
3 Institute of Physics, Jagiellonian University, Cracow, Poland
Received: 2004.02.23, Accepted: 2005.08.10, Published online first: 2006.02.13
Abstract
Introduction: Taurine chloramine (TauCl) and taurine bromamine (TauBr) are the main haloamines produced by activated
neutrophils. TauCl exerts both anti-inflammatory and microbicidal activities. Clinical studies showed that TauCl may be use-
ful as an antimicrobial agent in the local treatment of infections. Much less is known about TauBr. Circumstantial evidence
suggests that Propionibacterium acnes (PA) has a role in the inflammation of acne. Available topical therapies include antimi-
crobial agents which reduce total PA numbers and anti-inflammatory agents which suppress activity of the cells present in
acne inflammatory lesions. In this study the bactericidal activities of TauBr and TauCl against PA and Staphylococcus epi-
dermidis (SE), as a control strain, were investigated. Moreover, the influence of these haloamines on the generation of reac-
tive oxygen species (ROS) by activated neutrophils was also tested.
Materials and Methods: TauBr and TauCl were prepared by reaction of taurine with HOBr and HOCl, respectively. The reac-
tion was monitored by UV absorption spectra. The bactericidal activities of TauBr and TauCl were determined by the pour-
plate method. The generation of ROS by neutrophils was determined by luminol chemiluminescence assay.
Results: In our experimental set-up, TauBr showed stronger antibacterial activity than TauCl. Interestingly, PA was signifi-
cantly more susceptible to TauBr than SE was. Moreover, TauBr at non-cytotoxic concentrations significantly reduced ROS
generation by neutrophils.
Conclusions: Since PA is considered to be an etiological agent in acne and ROS are closely correlated with the pathogenesis
of inflammatory skin diseases, the reported data suggest that TauBr may be a good candidate for the topical therapy for acne
vulgaris.
Key words: acne vulgaris, Propionibacterium acnes, Staphyloccocus epidermidis, taurine bromamine, N-bromotaurine, taurine
chloramine, N-chlorotaurine, bactericidal activity, anti-inflammatory properties.
Regulatory T cells: magic bullets for immunotherapy?
Oliver Frey1 and Rolf Bräuer2
1 Institute of Immunology, University Hospital, Friedrich Schiller University, Jena, Germany
2 Institute of Pathology, University Hospital, Friedrich Schiller University, Jena, Germany
Received: 2005.04.11, Accepted: 2005.05.10, Published online first: 2006.02.13
Abstract
In the past few years it has been become increasingly clear that T cells capable of actively suppressing immune responses are
thought to be in part responsible for the maintenance of peripheral self tolerance. In healthy rodents and humans, CD4+ T
cells constitutively expressing the interleukin (IL)-2 receptor α-chain (CD25) are able to exert such suppressive function in
vitro and in vivo. Despite great efforts in our understanding of the biology of such immunoregulatory T cells, there are still
certain points incompletely understood. Although some authors suggest that immunoregulatory cytokines such as IL-10 or
transforming growth factor-β are critical for the suppressive effect of these cells, this is controversial and the exact molecu-
lar nature and the targets of suppression are largely unknown. Thus far, until regulatory T cells can be used for diagnostic or
therapeutic purposes many questions have to be answered. In this review we summarize the current knowledge on the func-
tion and properties of this T cell subset and discuss their potential role in human autoimmune or chronic inflammatory dis-
eases.
Key words: T cell subset, regulatory T cell, autoimmune disease, inflammation, infection, immunoregulation
The effect of
Chlamydia trachomatis infection
of the prostate gland on the concentration of citric acid
Bożena Zdrodowska−Stefanow1, Iwona Ostaszewska−Puchalska2,
Jerzy Badyda3 and Zofia Galewska4
1 Department of Dermatology and Venerology, Medical University, Bia³ystok, Poland
2 Center for Sexual Transmitted Disease Research and Diagnostics, Bia³ystok, Poland
3 Outpatients Urology Clinic “Multimedica”, Bia³ystok, Poland
4 Department of Medical Biochemistry, Medical University, Bia³ystok, Poland
Received: 2005.03.30, Accepted: 2005.09.19, Published online first: 2006.02.13
Abstract
Introduction: The objective of the study was to assess the relationship between Chlamydia trachomatis (C.t.) infection of the
prostate and the concentration of citric acid.
Materials and Methods: The study involved 60 patients with chronic prostatitis (NIH III). Urethral swabs and expressed pro-
static secretions (EPS) were collected for analysis. The urethral swabs were tested for PMNs and the presence of Neisseria
gonorrhoeae and C.t., while the EPS were analyzed to determine PMN count, C.t., and citric acid concentration. The DFA or
LCR method was used for C.t. diagnosis. The concentration of citric acid was measured using the UV method.
Results: Inflammation of the prostate (PMNs ≥10/field) was diagnosed in 58.3% of the patients. C.t. infection was found in
20%, including 8.3% with only the urethra affected and 10% with only the prostate. One patient had both the urethra and
the prostate infected. A reduction in the concentration of citric acid in EPS was observed in 56.7% of the men. In 88.2% of
the patients, reduced citric acid concentration was accompanied by an elevated PMN count in the EPS. All patients with C.t.
infection of the prostate showed a reduced concentration of citric acid. In five patients with urethral infection, lack of a
decrease in this parameter was noted in one. In all the patients with chlamydial infection, irrespective of localization, a high
PMN count was observed in the EPS.
Conclusions: Determination of the concentration of citric acid in the prostatic fluid is a good indicator of prostatitis. C.t.
infection of the prostate gland is accompanied by a decrease in the concentration of citric acid.
Key words: Chlamydia trachomatis, citric acid, polymorphonuclear leukocyte, urethra, prostate gland, expressed prostatic
secretions.
P-selectin mediates adhesion of leukocytes, platelets,
and cancer cells in inflammation, thrombosis,
and cancer growth and metastasis
Ming Chen1 and Jian−Guo Geng2
1 Laboratory of Molecular Cell Biology, Institute of Biochemistry and Cell Biology, Shanghai Institutes
for Biological Sciences, Graduate School of Chinese Academy of Sciences, Shanghai 200031, China
2 Vascular Biology Center and Division of Hematology, Oncology and Transplantation, University of Minnesota
Medical School, Minneapolis, MN 55455, USA
Received: 2005.06.20, Accepted: 2005.12.20, Published online first: 2006.03.24
Abstract
Stimulated endothelial cells and activated platelets express P-selectin (CD62P), a member of the selectin family of cell adhe-
sion molecules, which interacts with P-selectin glycoprotein ligand-1 (PSGL-1, CD162) for leukocyte rolling on stimulated
endothelial cells and heterotypic aggregation of activated platelets onto leukocytes. Cross-linking of PSGL-1 by P-selectin
also primes leukocytes intracellularly for cytokine and chemoattractant-induced β2-integrin activation for firm adhesion of
leukocytes. Furthermore, P-selectin mediates heterotypic aggregation of activated platelets to cancer cells and adhesion of
cancer cells to stimulated endothelial cells. Here we provide a comprehensive summary of the functional roles and the bio-
logical importance of P-selectin-mediated cell adhesive interactions in the pathogeneses of inflammation, thrombosis, and
the growth and metastasis of cancers.
Key words: selectin, cell adhesion molecule, leukocytes, platelets, endothelial cells, cancer cells, inflammation, thrombosis,
cancer growth and metastasis.
Monoclonal and bispecific antibodies
as novel therapeutics
Evan P. Booy1, 2, Dina Johar1, 2, Srilekha Maddika1, 2, Hasan Pirzada1,
Mickey M. Sahib3, Iris Gehrke1, Shauna Loewen1, 2, Sherif F. Louis1,
Kamran Kadkhoda1, 2, Michael Mowat1, 2 and Marek Los1, 2, 4
1 Manitoba Institute of Cell Biology, CancerCare Manitoba, University of Manitoba, Winnipeg, Canada
2 Department of Biochemistry and Medical Genetics, University of Manitoba, Winnipeg, Canada
3 Department of Oral Biology, University of Manitoba, Winnipeg, Canada
4 Manitoba Institute of Child’s Health, University of Manitoba, Winnipeg, Canada
Received: 2005.09.06, Accepted: 2005.11.30, Published online first: 2006.03.24
Abstract
Gene amplification, over-expression, and mutation of growth factors, or the receptors themselves, causes increased signal-
ing through receptor kinases, which has been implicated in many human cancers and is associated with poor prognosis.
Tumor growth has been shown to be decreased by interrupting this process of extensive growth factor-mediated signaling by
directly targeting either the surface receptor or the ligand and thereby preventing cell survival and promoting apoptosis.
Monoclonal antibodies have long been eyed as a potential new class of therapeutics targeting cancer and other diseases.
Antibody-based therapy initially entered clinical practice when trastuzumab/Herceptin became the first clinically approved
drug against an oncogene product as a well-established blocking reagent for tumors with hyperactivity of epidermal growth
factor signaling pathways. In the first part of this review we explain basic terms related to the development of antibody-based
drugs, give a brief historic perspective of the field, and also touch on topics such as the “humanization of antibodies” or cre-
ation of hybrid antibodies. The second part of the review gives an overview of the clinical usage of bispecific antibodies and
antibodies “armed” with cytotoxic agents or enzymes. Further within this section, cancer-specific, site-specific, or signaling
pathway-specific therapies are discussed in detail. Among other antibody-based therapeutic products, we discuss: Avastin
(bevacizumab), CG76030, Theragyn (pemtumomab), daclizumab (Zenapax), TriAb, MDX-210, Herceptin (trastuzumab),
panitumumab (ABX-EGF), mastuzimab (EMD-72000), Erbitux (certuximab, IMC225), Panorex (edrecolomab), STI571,
CeaVac, Campath (alemtuizumab), Mylotarg (gemtuzumab, ozogamicin), and many others. The end of the review deliber-
ates upon potential problems associated with cancer immunotherapy.
Key words: Bexxar, Gleevec, Imantib, Mitumomab, Tositumomab, Trastuzumab.
Novel therapies of multidrug-resistantPseudomonas aeruginosa and
Acinetobacter spp
.
infections: the state of the art
Marta Wróblewska
Department of Medical Microbiology, Medical University of Warsaw, and Infection Control Team, National Tuberculosis
and Lung Diseases Research Institute, Warsaw, Poland
Received: 2005.07.05, Accepted: 2005.12.20, Published online first: 2006.03.24
Abstract
Gram-negative non-fermenting bacilli, particularly Pseudomonas aeruginosa and Acinetobacter spp., are important oppor-
tunistic pathogens in hosppitalized patients, contributing to their morbidity and mortality. Recently, a rapid increase in fre-
quency of multidrug-resistant clinical strains is being recorded, making the available therapeutic options very limited. Apart
from the development of novel classes of antimicrobials, there is renewed interest in the use of old agents or new combina-
tions of available drugs. Numerous in vitro investigations have been reported on the efficacy of different antimicrobials; how-
ever, they should be evaluated in experimental infection models and clinical trials. Novel approaches are being investigated,
such as inhibition of virulence factor expression by pathogens or inhibition of their metabolic pathways. The use of bacte-
riophages, particularly those genetically modified, remains an alternative option in the therapy of infections caused by mul-
tidrug-resistant strains. Several vaccines against P. aeruginosa are under development. Apart from therapy with antimicro-
bial agents, eradication of outbreaks comprises implementation of strict infection control measures and prudent use of
antimicrobials.
Key words: Pseudomonas, Acinetobacter, vaccines, antimicrobial therapy.
The distinctive role of small heat shock proteins
in oncogenesis
Krzysztof Laudanski1 and Dorota Wyczechowska2
1 Department of Medicine, Griffin Hospital, Derby, CT 06418, USA
2 Department of Medicinal Chemistry, Medical University of £ódŸ, Poland
Received: 2005.12.25, Accepted: 2006.02.06, Published online first: 2006.03.28
Abstract
Recently, the role of small heat shock proteins (HSPs) has been widely recognized in cancer research. Small HSPs are tumor-
protective via numerous, independent mechanisms such as: oxidative stress, protection preventing protein denaturation,
anti-apoptotic activity, and likely direct suppression of the immune system. However, it is unclear whether they play any role
in the initial steps in carcinogenesis. This article seeks to familiarize the reader with general characteristics of small HSPs
(especially HSP-27, αA/B-crystallins), their tissue distribution with special regard on expression in malignant specimens, and
biological properties of intracellular HSP-27 and αA/B-crystallins promoting tumor genesis and growth. A separate chapter
describes immunomodulatory characteristics of extracellular HSP-27 with special emphasis on their plausible effect on the
neoplasm development.
Key words: small heat shock proteins, αA-crystallin, αB-crystallin, HSP-27, carcinogenesis, neoplasm, immune system.
Toll-like receptors types 2 and 6
and the apoptotic process in human neutrophils
Ewa Jabłońska1, Magdalena Marcińczyk1 and Jakub Jabłoński2
1 Department of Immunology, Medical University of Bia³ystok, Poland
2 Department of Toxicology, Medical University of Bia³ystok, Poland
Received: 2005.06.10, Accepted: 2006.02.08, Published online first: 2006.03.28
Abstract
Introduction: Neutrophils (PMN) apoptosis plays an important role in limiting the last phase of inflammatory processes. It is
unknown whether Toll-like receptor (TLR)2 acts independently or together with TLR6 in this process.
Materials and Methods: The aim of this study was to estimate the relationship between the expressions of TLR2 and TLR6
and the apoptosis of human neutrophils in physiological conditions. We investigated the influence of recombinant human
interleukin (IL)-18 and N-formyl-metionyl-leucyl-phenylalanine (fMLP) on the relationships between these receptors and
neutrophil apoptosis.
Results: Our results showed that after 4-h incubation, the percentage of apoptotic PMNs significantly increased compared
with PMN counts before incubation. The stronger expression of TLR2 on the neutrophils suggests that this receptor con-
tributes more significantly to the induction of PMN apoptosis than does TLR6. We also demonstrated an influence of recom-
binant human IL-18 (rhIL-18) on the expression of TLR6, whereas this effect was not observed in the expression of TLR2.
We observed that both rhIL-18 and fMLP inhibited the apoptosis of PMNs and that rhIL-18 had a stronger effect than fMLP.
Conclusions: The obtained results suggest that not only TLR2, but also TLR6 plays an important role in the regulation of the
apoptosis of PMNs. Changes in the expression of TLR6 and inhibition of apoptosis of PMNs by rhIL-18 seem to confirm the
vital role this receptor and of rhIL-18 in regulating the survival of these cells. These data can be useful in developing meth-
ods to regulate PMN apoptosis in conditions associated with their excessive and unfavorable activation.
Key words: neutrophils (PMNs), TLR2, TLR6, apoptosis, IL-18, fMLP
The migration of bone marrow-derived non-hematopoietic
tissue-committed stem cells is regulated
in an SDF-1-, HGF-, and LIF-dependent manner
Magda Kucia1, Wojtek Wojakowski2, Ryan Reca1, Bogdan Machalinski3,
Jolanta Gozdzik4, Marcin Majka4, Jarek Baran4, Janina Ratajczak1
and Mariusz Z. Ratajczak1, 4
1 Stem Cell Biology Program at James Graham Brown Cancer Center and Department of Medicine,
University of Louisville, Louisville, KY 40202, USA
2 Third Division of Cardiology, Silesian School of Medicine, Katowice, Poland
3 Pomeranian Medical School of Medicine, Szczecin, Poland
4 European Stem Cell Therapeutic Excellence Center, Medical College, Jagiellonian University, Cracow, Poland
Received: 2005.08.18, Accepted: 2005.12.20, Published online first: 2006.03.28
Abstract
Introduction: Recently we identified in bone marrow (BM) by employing chemotactic isolation to SDF-1 gradient combined
with real time RT-PCR analysis a mobile population of CXCR4+ BM mononuclear cells that express mRNA for various
markers of early tissue-committed stem cells (TCSCs). In this study we evaluated whether TCSCs respond to other moto-
morphogens, such as hepatocyte growth factor (HGF) and leukemia inhibitory factor (LIF).
Materials and Methods: We again employed chemotactic isolation combined with real-time RT-PCR analysis to assess
whether murine and human BM contain TCSCs that respond to HGF and LIF gradients. We also evaluated expressions of
HGF and LIF in damaged organs.
Results: We noted that the number of TCSCs is highest in BM from young (1- to 2-month-old) mice and decreases in 1-year-
-old animals. Murine and human TCSCs 1) respond to HGF and LIF gradients in addition to an SDF-1 gradient, 2) reside
in populations of BM-derived non-hematopoietic CD45– cells, and 3) are released (mobilized) from BM into the peripheral
blood (PB) during tissue injury (e.g. after partial body irradiation).
Conclusions: These findings further support our theory of the BM as a “hideout” for TCSCs and we suggest that their pres-
ence in BM tissue should be considered before experimental evidence is interpreted simply as transdifferentiation/plasticity
of hematopoietic stem cells. Since we demonstrated that not only SDF-1, but also HGF and LIF are upregulated in damaged
tissues, we postulate that CXCR4+ c-Met+ LIF-R+ TCSC could be mobilized from the BM into the PB, from which they are
subsequently chemoattracted to damaged organs, where they play a role in tissue repair/regeneration.
Key words: CXCR4-SDF-1, c-MET-HGF, LIF-R-LIF, stem cell plasticity, stem cell mobilization, regeneration.
Elevated TGF-ββ1 concentration in bronchoalveolar
lavage fluid from patients with primary lung cancer
Joanna Domagała−Kulawik1, Grażyna Hoser2, Aleksandra Safianowska1,
Hanna Grubek−Jaworska1 and Ryszarda Chazan1
1 Department of Pneumonology and Allergology, Warsaw Medical School, Poland
2 Department of Clinical Cytology, Medical Center of Postgraduate Education, Warsaw, Poland
Received: 2005.06.14, Accepted: 2005.12.18, Published online first: 2006.03.28
Abstract
Introduction: Transforming growth factor (TGF)-β is one of numerous inhibitory factors produced by cancer cells that regu-
late antitumor immunity. The aim of this study was to evaluate TGF-β1 levels and lymphocyte subsets in the broncholaveo-
lar lavage fluid (BALF) of patients with primary lung cancer and to analyze the interdependence of these parameters.
Materials and Methods: BALF samples were collected from 38 patients with primary lung cancer prior to treatment and from
23 healthy volunteers. Concentrations of TGF-β1 were measured in two independent lots of samples using a commercially
available sandwich ELISA kit after concentration of the supernatants. Differential cell counts in the BALF were performed
on slides stained with the May Grünwald Giemsa method. Flow cytometry with monoclonal antibodies was applied for lym-
phocyte phenotyping.
Results: A higher level of TGF-β1 in the BALF of patients compared with the healthy subjects was observed in both lots of
samples (3.23±2.96 pg/ml vs. 1.05±0.95 pg/ml, p<0.05, and 16.1±19.3 pg/ml vs. 10.1±11.1 pg/m,, respectively, difference not
significant). There was significant positive correlation of the TGF-β1 level with the proportion of lymphocytes and negative
correlation with both the proportion of macrophages and the percentage of cytotoxic and activated T lymphocytes.
Conclusions: Our findings confirmed that TGF-β takes part in the local response in the course of primary lung cancer.
Key words: lung cancer, bronchoalveolar lavage, TGF-β, lymphocyte
ETS transcription factors and regulation of immunity
Sarah Gallant and Gary Gilkeson
Department of Medicine, Medical University of South Carolina, Charleston, SC 29425, USA
Received: 2005.10.25, Accepted: 2005.12.30, Published online first: 2006.05.02
Abstract
The ETS family is a diverse group of transcription factors that control the expressions of genes that participate in an array
of cellular activities, ranging from mitosis to apoptosis. As a consequence of regulating these processes, most ETS factors are
oncogenic. However, there is growing evidence that ETS factors are also essential to regulation of the immune system. Of
the 29 recognized ETS factors, nine are known to regulate genes involved in immunity, including Ets1, Ets2, GABP, Fli1,
Elf1, MEF, ESE1, PU.1, and SpiB. These ETS factors typically activate the transcription of genes associated with pathogen
and tumor defense, but several also demonstrate ability to repress transcription. Ets1 and PU.1 appear to have the greatest
impact on immunity, primarily through their control of immune cell development. Alterations of Fli1 and Elf1 expression
are associated with autoimmunity, emphasizing the role of ETS factors as not only positive, but also negative regulators of
immunity. This review summarizes the roles of ETS factors in development of the immune system, defense against pathogens
and malignancies, and self-tolerance.
Key words: ETS, Ets1, Ets2, GABP, Fli1, Elf1, MEF, ESE1, PU.1, SpiB, transcription factor, immunity
Impact of fetal-maternal tolerance
in hematopoietic stem cell transplantation
Takanori Teshima1, Ken−ichi Matsuoka2 and Tatsuo Ichinohe3
1 Center for Cellular and Molecular Medicine, Kyushu University Hospital, 3-1-1 Maidashi, Higashi-ku,
Fukuoka 812-8582, Japan
2 Biopathological Science, Okayama University Graduate School of Medicine and Dentistry, 2-5-1 Shikata-cho,
Okayama 700-8558, Japan
3 Department of Hematology/Oncology, Graduate School of Medicine, Kyoto University, 54 Shogoin Kawahara-cho,
Sakyo-ku, Kyoto 606-8507, Japan
Received: 2006.01.05, Accepted: 2006.02.06, Published online first: 2006.05.02
Abstract
Allogeneic hematopoietic stem cell transplantation (HSCT) is known to cure various hematological disorders; however, its
widespread use is limited due to a lack of histocompatible donors. Reciprocal cell traffic between the mother and fetus dur-
ing pregnancy gives rise to postpartum fetal-maternal lymphohematopoietic microchimerism, which is frequently detected in
the blood or tissue of healthy individuals. Studies in clinical and experimental transplantation provide evidence that expo-
sure to non-inherited maternal antigens (NIMAs) during pregnancy may result in long-lasting fetomaternal microchimerism
and tolerance induction. Studies of HLA-mismatched HSCT have suggested a relatively lower incidence of severe graft-ver-
sus-host disease (GVHD) after transplantation from a NIMA-mismatched donor. Studies using a mouse model have also
demonstrated a “child-to-mother” bone marrow transplantation from an NIMA-exposed donor to reduce the morbidity and
mortality of GVHD in an antigen-specific manner while preserving the graft-versus-leukemia effects and favoring the
immune reconstitution, thus resulting in a marked improvement in outcome after HSCT. Prospective clinical studies are
therefore warranted to confirm these beneficial effects of fetal-maternal tolerance in allogeneic HSCT.
Key words: HSCT, GVHD, non-inherited maternal antigen.
Modulation of apoptosis signaling for cancer therapy
Simone Fulda and Klaus−Michael Debatin
University Children’s Hospital, Ulm, Germany
Received: 2005.11.14, Accepted: 2006.02.06, Published online first: 2006.05.02
Abstract
Apoptosis, the cell’s intrinsic death program, plays a central role in regulating tissue homeostasis. Also, most cytotoxic ther-
apies used for cancer treatment, such as chemotherapy, γ-irradiation, suicide genes, or immunotherapy, predominantly act
by triggering apoptosis in target cells. Thus, understanding the molecular events that regulate apoptosis and how tumor cells
evade apoptotic deletion have provided a paradigm to link cancer genetics and response to cancer therapy. Therefore,
insights into the mechanisms regulating drug-induced apoptosis provide rational targets for novel therapeutic interventions.
Key words: apoptosis, cancer therapy, resistance.
Haemophilus influenzae type b
and pertussis vaccinations in preterm infants
Janusz Piotr Sikora, Danuta Chlebna−Sokół, Iwona Ligenza and Anna Sikora
Department of Pediatric Propedeutics and Bone Metabolic Diseases, Medical University of £ódŸ, University Clinical
Hospital No. 4, £ódŸ, Poland
Received: 2005.07.15, Accepted: 2006.12.08, Published online first: 2006.05.02
Abstract
Introduction: The aim of the study was to evaluate the serological efficacy of Hiberix and Infanrix-DTPa vaccines in preterm
infants.
Materials and Methods: The results of the investigation of 61 preterm infants immunized three times (primary vaccination)
with Hiberix and Infanrix-DTPa at 6-week intervals are presented. Of the 61 children, 17 were additionally immunized with
a booster dose of these vaccines. Postvaccinal response to these immunizations was evaluated by means of an immunoenzy-
matic method.
Results: We observed a significant increase in protective postvaccinal antibody titers against Haemophilus influenzae type b
(Hib) and Bordetella pertussis after the primary vaccination compared with the initial antibody levels (p<0.05). A significant
increase (p<0.0002) in protective antibody titers after the booster dose of Hiberix compared with the primary vaccination
was also noted. No correlations between birth weight, gestational age, and the achieved levels of postvaccinal anti-polyribo-
sylribitol phosphate of Hib and of anti-pertussis toxin and anti-filamentous hemagglutinin of B. pertussis antibodies after the
primary vaccination or booster dose were found. After the booster dose, all the preterm infants responded with the produc-
tion of protective postvaccinal antibody titers against Hib and B. pertussis.
Conclusions: Due to the very good immunogenicity of the vaccines against Hib studied, inclusion of this immunization should
be proposed in the obligatory vaccination schedule in Poland, especially in preterm infants. An additional immunization (i.e.
a second booster dose) of Polish children with acellular pertussis (DTPa) vaccine is necessary to protect them from decreas-
ing protective anti-pertussis antibody titers in early childhood.
Key words: Hib, pertussis, vaccination, preterm infants.
Association of CD45dimVLA-4+ cells
with the NKT cell lineage and their selective expression
of IL-13, IP-15, and CCR3 transcripts
Agata Matejuk1 and Michael Afentoulis2
1 Department of Pathology, Oregon Health and Science University, Portland, OR 97239, USA
2 Neuroimmunology Research, Veterans Affairs Medical Center, Portland, OR 97239, USA
Received: 2005.11.17, Accepted: 2006.02.10, Published online first: 2006.05.31
Abstract
Introduction: Estrogen (E2) was shown to prevent experimental autoimmune encephalomyelitis (EAE) and to produce
a novel population of regulatory CD45dimVLA-4+ cells. Although their appearance was dependent upon an elevated hor-
monal level, E2 was not required for their production, as they also were induced by immunization with Mycobacterium tuber-
culosis as a component of complete Freund’s adjuvant.
Materials and Methods: Molecular techniques, including ribonuclease protection assays and quantitative RT-PCR, were used
to provide further characterization of CD45dimVLA-4+ cells. Moreover, we determined the developmental requirements of
the CD45dimVLA-4+ cells using genetically modified mice and extensive flow cytometry analysis.
Results: Characterization of CD45dimVLA-4+ mRNA profile revealed highly elevated levels of CD16, CD44, CCR3, IP-15,
and IL-13 transcripts compared with their CD45highVLA-4+ counterparts. Furthermore, we found up-regulation of anti-
-apoptotic bcl-w and bcl-xl genes and transcripts encoding the TCRα and CD8α homodimer. The production of
CD45dimVLA-4+ cells was evident in nude mice and in MHC class II- and β2-microglobulin, but not in CD1-deficient mice,
suggesting a crucial role for CD1 in their induction.
Conclusions: These findings suggest that CD45dimVLA-4+ cells might resemble natural killer T cells and imply possible roles
for IL-13 and IP-15 in the protective function of CD45dimVLA-4+ cells. A better understanding of how these cells, also occur-
ring naturally during pregnancy, suppress the harmful immune response of EAE may lead to novel therapeutic approaches
to combat multiple sclerosis and other autoimmune diseases.
Key words: autoimmunity, EAE/MS, regulatory cells, adhesion molecules, chemokines/cytokines, pregnancy tolerance.
Characterization and serological classification
of O-specific polysaccharide of
Proteus mirabilis TG 276-90
from
Proteus serogroup O34
Katarzyna Kołodziejska1, Małgorzata Siwińska1, Krystyna Zych1, Antoni Różalski2
and Zygmunt Sidorczyk1
1 Department of General Microbiology, Institute of Microbiology and Immunology, University of £ódŸ, Poland
2 Department of Immunobiology of Bacteria, Institute of Microbiology and Immunology, University of £ódŸ, Poland
Received: 2005.06.27, Accepted: 2005.12.08, Published online first: 2006.05.31
Abstract
Introduction: Gram-negative bacteria of the genus Proteus from the family Enterobacteriaceae are currently divided into the
five species P. mirabilis, P. vulgaris, P. penneri, P. hauseri, and P. myxofaciens and three unnamed Proteus genomospecies 4,
5, and 6. They are important facultative human and animal pathogens which, under favorable conditions, cause mainly
intestinal and urinary tract infections, sometimes leading to serious complications such as acute or chronic pyelonephritis
and the formation of bladder and kidney stones. In this study we report on the serological properties of the lipopolysaccha-
ride (LPS) of P. mirabilis TG 276-90, whose O-polysaccharide chemical structure was described earlier.
Materials and Methods: LPS and alkali-treated LPS of a few serologically related Proteus strains and O-antisera against
P. mirabilis TG 276-90 and CCUG 4669 (O34) were used. Serological characterization of P. mirabilis TG 276-90 O-specific
polysaccharide was done using enzyme immunosorbent assay, passive immunohemolysis test (PIH), inhibition of these tests,
SDS/PAGE and Western blot techniques, absorption of rabbit polyclonal O-antisera, and repeated PIH test.
Results: Structural and serological investigations showed that the O-polysaccharides of P. mirabilis TG 276-90 and P. vulgaris
O34 are identical and that their LPSs differ only in epitopes in the core part. Therefore these two strains could be classified
into the same Proteus O34 serogroup.
Conclusions: The serological data showed that the β-D-GalpNAc-(1→4)-α-D-GalpNAc disaccharide is an important epitope
of the P. mirabilis TG 276-90 and P. vulgaris O34 LPSs, shared by the P. mirabilis O16 and P. vulgaris TG 251 LPSs. It is
responsible for cross-reactions with P. mirabilis TG 276-90 and P. vulgaris O34 O-antisera.
Key words: Proteus mirabilis, Proteus vulgaris, lipopolysaccharide, O-serogroup, O-antigen.
TNF-αα priming effect on polymorphonuclear
leukocyte reactive oxygen species generation
and adhesion molecule expression
in hemodialyzed patients
Jacek Rysz1, Maciej Banach2, Robert A. Stolarek1, Jarosław Pasnik3,
Aleksandra Ciałkowska−Rysz4, Leszek Markuszewski5 and Zbigniew Baj6
1 2nd Department of Family Medicine, Medical University Hospital No. 2, Medical University of £ódŸ, Poland,
2 Department of Cardiac Surgery; Medical University Hospital No. 3, Medical University of £ódŸ, Poland
3 Department of Pediatrics and Clinical Immunology; Medical University of £ódŸ, Poland
4 Department of Palliative Care, Medical University of £ódŸ, Poland
5 Department of Interventional Cardiology, Cardiodiabetology and Cardiac Rehabilitation, Medical University Hospital
No. 2, Medical University of £ódŸ, Poland
6 Department of Pathophysiology and Clinical Immunology, Medical University of £ódŸ, Poland
Received: 2005.04.25, Accepted: 2005.12.08, Published online first: 2006.05.31
Abstract
Introduction: The study aimed to assess reactive oxygen species generation and the expressions of some surface antigens on
polymorphonuclear leukocytes (PMNs) in patients on regular hemodialysis (HD) treatment.
Materials and Methods: The respiratory burst of PMNs was determined with luminol-dependent chemiluminescence (CL) in
resting cells and following N-formyl-methionyl-leucyl-phenylalanine (fMLP), phorbol 12-myristate 13-acetate (PMA), or
opsonized zymosan (OZ) stimulation and expressed in arbitrary CL units times assay-time (aU×min). The expressions of
CD11b/CD18, CD10, and CD13 receptors were determined with flow cytometry.
Results: Basal PMN CL was increased in HD patients to up to 1285±129 aU×min compared with 895±88 aU×min in healthy
controls (p<0.05). The CL of unprimed PMNs increased after fMLP stimulation from 3085±746 to 4529±808 aU×min, and
after OZ stimulation from 12945±1296 to 14678±1355 aU×min. PMA-stimulated CL of PMNs was similar to control values.
The oxidative burst in PMNs from HD patients and healthy controls was similar in response to TNF-α alone. The CL of
TNF-α-primed PMNs in HD patients was significantly lower than CL measured in healthy controls (p<0.05). The expres-
sions of CD10 and CD13 metalloproteinase receptors were also increased (p<0.05). Although CD11b expression was sig-
nificantly increased at rest and after fMLP stimulation, the expression of another β-integrin heterodimer compound, CD18,
was not increased.
Conclusions: These results provide evidence that TNF-α priming of PMNs is down-regulated in HD patients despite consti-
tutive up-regulation of resting cytotoxicity and enhanced expression of adhesion and metalloproteinase receptors.
Key words: hemodialysis, neutrophils, chemiluminescence, TNF-α, CD11b, CD18.
The use of the Congo red-related dye DBACR
to recognize the heavy chain-derived abnormality
of myeloma immunoglobulins
Paweł Spólnik1, Leszek Konieczny1, Barbara Piekarska1, Janina Rybarska1,
Barbara Stopa1, Grzegorz Zemanek1, Anna Drozd1, Marcin Król2*, Irena Roterman2,
Teresa Wolska−Smoleń3 and Aleksander B. Skotnicki3
1 Institute of Medical Biochemistry, Medical College, Jagiellonian University, Kraków, Poland
2 Department of Bioinformatics and Telemedicine, Medical College, Jagiellonian University, Kraków, Poland
3 Department of Hematology, Medical College, Jagiellonian University, Kraków, Poland
* Present address: Biomolecular Modelling Laboratory, Cancer Research UK London Research Institute Lincoln’s Inn
Fields Laboratories, London WC2A 3PX, UK
Received: 2005.06.14, Accepted: 2005.12.23, Published online first: 2006.05.31
Abstract
Introduction: The aim of this study was to differentiate heavy and light chain-derived instability of monoclonal myeloma
immunoglobulins by complexation of matched supramolecular dyes. These are composed of several micellar pieces of self-
-assembled dye molecules which may penetrate the protein interior of the binding locus with polypeptide chains. These dyes
were used to elicit, by precipitation, the postulated higher aggregation tendency of the heavy chain derived from its higher
hydrophobicity.
Materials and Methods: Agarose gel electrophoresis was used to create conditions for dye complexation and to reveal the pre-
cipitation.
Results: Congo red derivatives with aromatic ring substitutes, BACR and DBACR, of increased penetrating capability were
chosen to provoke the precipitation of abnormal immunoglobulins by displacing association-prone polypeptide chains from
the protein interior.
Conclusions: The results of this study confirm the heavy chain-related propensity of some monoclonal immunoglobulins to
aggregate and precipitate. The simplicity of the technique may improve clinical diagnosis and facilitate predictions of disease
complications.
Key words: multiple myeloma, immunoglobulin aggregation, monoclonal immunoglobulins, Congo red, bis-azo dyes.
Nitric oxide production by pulmonary leukocytes
from induced sputum in patients with asthma
and its effect on epithelial cell viability
Małgorzata Bieńkowska−Haba1, Jerzy Liebhart2 and Monika Cembrzyńska−Nowak1
1 Laboratory of Virology, Institute of Immunology and Experimental Therapy, Polish Academy of Science, Wroc³aw, Poland
2 Department of Internal Medicine and Allergology, Wroclaw Medical University, Wroc³aw, Poland
Received: 2005.06.01, Accepted: 2006.02.06, Published online first: 2006.05.31
Abstract
Introduction: Nitric oxide (NO) is one of many factors potentially involved in lung remodeling in asthma. The aim of the study
was to assess the effect of pulmonary leukocytes from patients with bronchial asthma on alveolar epithelial cell damage in
relation to NO production.
Materials and Methods: Induced sputum samples were obtained from 25 patients with bronchial asthma and 10 healthy vol-
unteers. Twelve asthmatics were on inhaled corticosteroid treatment and 13 were corticosteroid free. Type II-like alveolar
epithelial (A549) cells were cultured for 48 h in the presence of cell-free media from a 24-h culture of leukocytes obtained
from the induced sputa (IS-Su). The level of NO was measured in supernatants from the cell cultures and the viability of the
A549 cells was established.
Results: The levels of NO in IS-Su from corticosteroid-free asthmatics were significantly higher (p=0.001) than those in
IS-Su from healthy controls. Furthermore, NO production by A549 cells exposed to IS-Su from steroid-free asthmatics
(group A) was significantly higher than that from asthmatics on corticosteroid therapy (group cA) as well as from healthy
controls (p=0.01 and p=0.001, respectively). Lower viability of the epithelial cells exposed to IS-Su was observed in group
A compared with controls (median: 72% vs. 97.5%; p<0.001). In addition, a negative correlation (RS=–0.706, p<0.001) was
found between the levels of NO produced by pulmonary leukocytes and the viability of epithelial cells.
Conclusions: The results suggest that in the course of asthma, pulmonary leukocytes may interact with alveolar epithelial cells
by inducing an excessive production of NO which, in turn, may contribute to epithelium impairment.
Key words: nitric oxide, asthma, lung remodeling, pulmonary leukocytes, alveolar epithelial cells.
Molecular addresses of tumors:
selection by
in vivo phage display
Xiao−Bo Li1, 2, Hermann J. Schluesener2 and Shun−Qing Xu1
1 MOE Key Lab of Environment and Health, Institute of Environmental Health, Tongji Medical College,
Huazhong University of Science and Technology, Wuhan, P. R. China
2 Institute of Brain Research, Tuebingen, Germany
Received: 2005.04.24, Accepted: 2006.05.02, Published online first: 2006.05.31
ABSTRACT
In vivo phage display has been used extensively to screen for novel targets of tumor therapy. Phage display peptide libraries
can express random peptides or protein fragments and the aim of phage display is to identify peptide molecules that bind
stably to a given target. Angiogenesis is essential to tumor development. Both blood and lymphatic vessels of tumors are dif-
ferent from those of normal tissues. Phage display has been used to analyze the structure and molecular diversity of tumor
vasculature and to select tumor-specific antigens which have revealed stage- and type-specific markers of tumor blood ves-
sels. Furthermore, peptides identified by in vivo phage display also work as vehicles to transport cargo therapeutic reagents
to tumors. These peptides and their corresponding cellular proteins and ligands may provide molecular tools to selectively
target the addresses of tumors and their pathological blood vessels and might increase the efficacy of therapy while decreas-
ing side effects.
Key words: phage display, tumor therapy, angiogenesis, tumor antigen
Deconstructing B cell tolerance to basement membranes
Mary H. Foster, Ying Zhang and Amy G. Clark
Departments of Medicine and Research Service, Duke University and Durham Veterans Affairs Medical Centers,
Durham, NC 27710, USA
Received: 2006.03.03, Accepted: 2006.05.02, Published online first: 2006.07.10
Abstract
Basement membrane antigens are frequent targets of autoantibody attack in systemic and organ-restricted autoimmunity.
These specialized and highly organized matrices are composed of multiple components with restricted tissue distributions
and limited epitope exposure. To dissect mechanisms controlling humoral autoimmunity to nephritogenic basement mem-
brane antigens, we developed autoantibody transgenic models. In mice bearing the LamH Ig transgene encoding B cell
receptors specific for laminin, autoreactive B cells are readily generated but actively regulated in vivo. In this model, anti-
-laminin B cells are immunologically censored by mechanisms that include central deletion, κ light-chain editing, and anergy.
Tolerance is maintained when the transgene is established in MRL and BXSB genetic backgrounds with inherited autoim-
mune susceptibility, and despite provocation with potent environmental stimulants. Collectively, these studies indicate that
the pathogenic anti-laminin reactivity characteristic of systemic lupus is tightly regulated. A novel anti-collagen transgenic
model is used to assess the tolerogenesis of a structurally distinct pathogenic basement membrane epitope and to determine
if reactivity to putative cryptic epitopes targeted in organ-restricted disease is regulated. These studies should provide insight
into the molecular mechanisms controlling basement membrane autoreactivity and ultimately facilitate the development of
novel strategies to inactivate autoreactive cells and treat autoimmune disease.
Key words: B cell tolerance, basement membrane, autoantibody, laminin.
Corresponding author: Mary H. Foster, M.D., De
The role of the lysophospholipid sphingosine 1-phosphate
in immune cell biology
Henrik von Wenckstern*, Karsten Zimmermann* and Burkhard Kleuser
Institute of Pharmacy, Pharmacology and Toxicology, Freie Universität Berlin, Germany
Received: 2006.03.14, Accepted: 2006.05.02, Published online first: 2006.07.10
Abstract
Sphingosine 1-phosphate (S1P) has been shown to be a bioactive lipid mediator intimately involved in mediating a variety of
immunological processes. In particular, S1P regulates lymphocyte cell trafficking between the lymphatic system and the
blood. The lysophospholipid signals mainly through five related G protein-coupled receptor subtypes, termed S1P1 to S1P5.
S1P1 seems to play an essential role in cell trafficking, as this receptor subtype promotes the egress of T and B cells from sec-
ondary lymphatic organs. This S1P1-mediated migratory response is a consequence of different S1P levels in the serum and
lymphatic organs. In addition to its direct effects on lymphocyte motility, S1P strengthens cell barrier integrity in sinus-lin-
ing endothelial cells, thereby reducing lymphocyte egress out of lymph nodes. Furthermore, S1P modulates cytokine profiles
in T and dendritic cells, resulting in an elevated differentiation of T helper-2 cells during the T cell activation process. It is
of interest that the mode of molecular action of the novel immunomodulator FTY720 interferes with the signaling of S1P.
After phosphorylation, FTY720 shares structural similarity with S1P, but in contrast to the natural ligand, phosphorylated
FTY720 induces a prolonged internalization of S1P1, resulting in an impaired S1P-mediated migration of lymphocytes.
Key words: sphingolipids, sphingosine 1-phosphate, FTY720, immunosuppression, T cell, dendritic cell.
Modifier effect of the Toll-like receptor 4 D299G
polymorphism in children with cystic fibrosis
Donald S. Urquhart1, Jennifer Allen2, Mohamed Elrayess3, Katy Fidler2, Nigel Klein2
and Adam Jaffé1
1 Portex Anaesthesia, Intensive Therapy and Respiratory Medicine Unit, Institute of Child Health,
and Great Ormond Street Hospital for Children NHS Trust, London, UK
2 Infectious Diseases and Microbiology Unit, Institute of Child Health, and Great Ormond Street Hospital
for Children NHS Trust, London, UK
3 Department of Cardiovascular Genetics, Rayne Institute, University College London, UK
Received: 2006.01.05, Accepted: 2006.05.04, Published online first: 2006.07.10
Abstract
Introduction: Clinical phenotype varies amongst cystic fibrosis (CF) patients with identical CF transmembrane regulator
(CFTR) genotype, suggesting genetic modifiers exist. One potential modifier is the Toll-like receptor 4 (TLR4) gene. TLR4
binds lipopolysaccharide (LPS), a constituent of Pseudomonas aeruginosa (PA), activating innate immunity and promoting
inflammation. TLR4 polymorphisms are associated with LPS-hyporesponsiveness and may be protective in CF due to
decreased inflammation.
Materials and Methods: DNA was extracted from blood of recruited CF subjects, and PCR performed to establish TLR4
D299G genotype. Case-notes were reviewed to obtain clinical data. Subjects possessing the TLR4 299G allele were com-
pared with age, sex, and CFTR genotype-matched wild-type (299DD) subjects and also with all controls.
Results: 100 subjects (mean age 8.9 years) were studied, with 11 299DG heterozygotes identified. On case-matched analyses,
no statistically significant differences between groups were found for mean±SEM rates of change of %predicted FEV1/year
(0.9±2.3 (DD) vs. –3.9±2.8 (DG), p=0.22), %predicted FEV1 (76±8 vs. 74±11), p=0.91), or z scores for height (–0.47±0.26
vs. –0.24±0.19, p=0.48) and weight (–0.01±0.22 vs. –0.29±0.27, p=0.44). Median±SE survival age at first PA isolation was
also not significantly different (3.5±2.1 vs. 6.5±2.4 years, p=0.29). No statistically significant differences were noted when
299DG heterozygotes were compared with all controls.
Conclusions: Potential reasons for absence of modifier effect include the basolateral location of TLR4 receptors on respira-
tory epithelium, or because inflammatory response to PA in the CF airway is so overwhelming that even a blunted response
(as suggested for the 299G allele) results in increased inflammation and lung damage.
Key words: cystic fibrosis, genotype-phenotype correlation, modifier genes, Toll-like receptor 4
Key factors in experimental mouse
hematopoietic stem cell transplantation
Dmitry Nevozhay and Adam Opolski
Department of Experimental Oncology, Institute of Immunology and Experimental Therapy, Polish Academy of Sciences,
Wroc³aw, Poland
Received: 2005.11.20, Accepted: 2006.04.03, Published online first: 2006.07.25
Abstract
The first mouse model of hematopoietic stem cell transplantation (HSCT) was developed more than 50 years ago. HSCT is
currently being widely used in a broad range of research areas, which include studies of the engraftment process, the patho-
genesis of graft-versus-host disease and possible ways of its treatment and prophylaxis, attempts to use the graft-versus-
-leukemia/tumor effect in treating hematological and oncological malignancies, cancer vaccine development, induction of
transplanted organ tolerance, and gene therapy. However, although this model is widely distributed, many laboratories use
different protocols for the procedure. There are a number of papers discussing different HSCT protocols in clinical work,
but no articles summarizing mouse laboratory models are available. This review attempts to bring together different details
about HSCT in the mouse model, such as the types of transplantation, possible pretreatment regimens and their combina-
tions, methods and sources of graft harvesting and preparation for the transplantation procedure, the influence of graft cell
dose and content on the engraftment process, the transplantation method itself, possible complications, symptoms and tech-
niques of their prophylaxis or treatment, as well as follow-up and engraftment assessment. We have also tried to reflect cur-
rent knowledge of the biology of the engraftment.
Key words: hematopoietic stem cell transplantation, mouse models, engraftment, chimerism
Structures and serology of the O-antigens
of Proteus strains classified into serogroup O17
and former serogroup O35
Agnieszka Torzewska1, Sebastian Grabowski1, Anna N. Kondakova2, Filip V. Toukach2,
Sof’ya N. Senchenkova2, Alexander S. Shashkov2, Nikolay P. Arbatsky2,
Yuriy A. Knirel2, Antoni Różalski1 and Wiesław Kaca3
1 Department of Immunobiology of Bacteria, Institute of Microbiology and Immunology, University of £ódŸ, Poland
2 N.D. Zelinsky Institute of Organic Chemistry, Russian Academy of Sciences, Moscow, Russia
3 Department of Microbiology, Institute of Biology, Œwiêtokrzyska Academy, Kielce, Poland
Received: 2005.08.05, Accepted: 2006.02.10, Published online first: 2006.07.25
Abstract
Introduction: Bacteria of the genus Proteus are facultative pathogens which commonly cause urinary tract infections. Based
on the serological specificity of the O-chain polysaccharide of the lipopolysaccharide (O-polysaccharide, O-antigen), strains
of P. mirabilis and P. vulgaris have been classified into 60 serogroups. Studies on the chemical structure and serological speci-
ficity of the O-antigens aim at the elucidation of the molecular basis and improvement of the serological classification of
these bacteria.
Materials and Methods: The O-polysaccharide was prepared by acetic acid degradation of the lipopolysaccharide isolated
from dried bacterial mass of each strain by hot phenol/water extraction. 1H- and 13C-NMR spectroscopy was used for struc-
tural studies. Serological studies were performed with rabbit O-antisera using enzyme immunosorbent assay, passive hemo-
lysis test, and the inhibition of reactions in these assays as well DOC-PAGE and Western blot.
Results: Four Proteus strains belonging to serogroups O17 and O35 were found to possess similar O-polysaccharide struc-
tures, in particular having the same carbohydrate backbone built up of tetrasaccharide repeating units. However, they differ
in the presence or absence of additional substituents, such as phosphoethanolamine in P. mirabilis O17 and glucose in P. pen-
neri O17, as well as in the pattern and degree of O-acetylation of various monosaccharide residues. Serological studies also
showed close relationships between the O-antigens studied.
Conclusions: Based on these data it is proposed to reclassify strain P. mirabilis PrK 61/57, formerly representing the O35
serogroup, into the serogroup O17 in the Kauffman-Perch classification system of Proteus.
Key words: Proteus vulgaris, Proteus mirabilis, O-antigen, O-polysaccharide structure, lipopolysaccharide, serological cross-
reactivity.
Circadian variations of histamine binding to lymphocytes
and neutrophils and skin reactivity to histamine
in atopic and healthy subjects
Teresa Żak−Nejmark, Iwona Anna Nowak and Maria Kraus−Filarska
Department of Internal Medicine and Allergology, Wroclaw Medical University, Wroc³aw, Poland
Received: 2005.08.18, Accepted: 2006.02.03, Published online first: 2006.07.25
Abstract
Introduction: Numerous pathophysiological conditions change during 24-hour periods. Histamine, the main mediator in aller-
gic reactions, exerts a multiplicity of pathophysiological actions through binding to specific receptors on effector cells.
Nocturnal exacerbation of symptoms occurs in many atopic diseases in which histamine is an important mediator. Nocturnal
wheezing is a very common symptom of asthma. The aim of this study was to determine whether the binding of (fluorescein-
-labeled) histamine to cells participating in allergic-inflammatory processes (lymphocytes, neutrophils) and skin reactivity to
histamine undergo circadian changes and to compare these phenomena in atopic asthmatic and healthy subjects.
Materials and Methods: Blood samples were collected at 8 am, 2 pm, 8 pm, 2 am, and 8 am the next day. Histamine skin-prick
tests were performed at the same times.
Results: It was found that skin reactivity to histamine (wheal, erythema) in healthy subjects underwent significant circadian
changes with acrophase at 8 am (wheal) or 8 pm (erythema), the lowest values being at night (2 am, p=0.017), in contrast to
atopics, in whom the highest reactivity was found at night (2 am, p=0.002). Significant differences in the binding of fluores-
cein-labeled histamine between day (8 am–2 pm) and night (2 am) were observed for lymphocytes (p=0.006) and neutrophils
(p=0.018).
Conclusions: In the asthmatic group these changes were not significant. Circadian changes in both the binding of histamine
by effector cells and skin reactivity to histamine were different in healthy and asthmatic subjects, and this may play a role in
the pathomechanism, course, and chronopharmacotherapy of atopic diseases.
Key words: histamine, circadian rhythms, lymphocytes, neutrophils, skin-prick test.
Matrix metalloproteinase and cytokine production
by bone marrow adherent cells
from multiple myeloma patients
Barbara Zdzisińska1, Adam Walter−Croneck2, Anna Dmoszyńska2
and Martyna Kandefer−Szerszeń1
1 Department of Virology and Immunology, Maria Curie-Sk³odowska University, Lublin, Poland
2 Department of Hematooncology and Bone Marrow Transplantation, Medical University of Lublin, Poland
Received: 2005.10.24, Accepted: 2006.02.23, Published online first: 2006.07.25
Abstract
Introduction: Cultures of bone marrow stromal cells derived from the bone marrow of multiple myeloma (MM) patients were
shown to exhibit several abnormalities compared with control cultures from healthy subjects. The aim of the study was to
examine whether cultures of bone marrow adherent cells, at low passage level, exhibit differences in matrix metallopro-
teinases (MMPs) and cytokine production compared with cultures from normal donors.
Materials and Methods: MMP production was evaluated by gel zymography and by ELISA in supernatants of serum-free cul-
tures of bone marrow adherent cells derived from 20 MM patients and 23 healthy controls. Spontaneous and lipopolysac-
charide (LPS)- or Newcastle disease virus (NDV)-induced cytokine release was assessed in the supernatants of the cultures
by the ELISA method.
Results: Both cultures produced MMP-1, -2, -3, and -9 under serum-free conditions; however, the levels of MMP-1 and
MMP-2 were significantly higher in cultures derived from MM patients, while MMP-3 was significantly higher in control cul-
tures. The level of MMP-9 was comparable in the cultures derived from MM patients and controls. All cultures produced
interleukin (IL)-10 and IL-11 spontaneously, but after LPS or NDV induction the levels of IL-10, IL-11, interferon α, and
tumor necrosis factor α, were significantly higher in the cultures derived from MM patients than in control cultures.
Conclusions: The results indicate that both the abnormalities in MMP production and the overproduction of cytokines (in
the presence of LPS or virus, which mimic inflammatory conditions) may be involved in bone destruction and tumor spread
in multiple myeloma.
Key words: multiple myeloma, bone marrow adherent cells, cytokines, matrix metalloproteinases
Shraga Segal (1939–2006)- Obituary
Noah Isakov
Induction and maintenance of self tolerance:
the role of CD4+CD25+ regulatory T cells
Kamalesh K. Bala and Kamal D. Moudgil
Department of Microbiology and Immunology, University of Maryland School of Medicine, Baltimore, MD 21201, USA
Received: 2006.04.14, Accepted: 2006.05.12, Published online first: 2006.10.06
Abstract
The immune system responds vigorously to invading pathogens (non-self, foreign), while remaining unresponsive (tolerant)
to the body’s own components and circulating constituents (self). This indifference to self components is a result of finely
orchestrated events of thymic negative selection (central tolerance) of developing T cells that are autoaggressive combined
with those operative in the periphery (peripheral tolerance) to control the activity of potentially autoreactive T cells that
escaped thymic tolerance. Recently, autoimmune regulator expressed in the thymus has been identified as a critical media-
tor of central tolerance towards tissue-specific antigens. In the periphery, a variety of regulatory T cells are involved in effect-
ing tolerance. There is immense interest and excitement about the newly identified subset of CD4+CD25+ T cells. This is
a unique subset of CD4+ T cells that bear CD25 (IL-2Rα chain) on the cell surface in the naïve state and express FoxP3 as
a unique marker. These cells suppress the activity of autoreactive effector T cells primarily via cell-cell contact. The defi-
ciency and/or altered function of CD4+CD25+ T cells is associated with autoimmunity. Mice deficient in FoxP3 (scurfy mice)
bear an autoimmune phenotype, and human males with mutations in the corresponding gene express the phenotype of wide
-spread autoimmunity, the immune dysregulation, polyendocrinopathy and enteropathy, and X-linked syndrome. In vitro
expansion of antigen-specific CD4+CD25+ T cells and their adoptive transfer into patients suffering from autoimmunity is
emerging as a promising new therapeutic approach for these debilitating disorders.
Key words: tolerance, regulatory T cells, CD4+CD25+ T cells, autoimmunity.
Thalidomide increases
in vitro sensitivity
of childhood acute lymphoblastic leukemia cells
to prednisolone and cytarabine
Krzysztof Czyżewski1, Agnieszka Zaborowska1 and Jan Styczyński2
1 Students’ Scientific Society, Department of Pediatric Hematology and Oncology,
Ludwik Rydygier Collegium Medicum of Bydgoszcz, Nicolaus Copernicus University, Bydgoszcz, Poland
2 Department of Pediatric Hematology and Oncology, Ludwik Rydygier Collegium Medicum of Bydgoszcz,
Nicolaus Copernicus University, Bydgoszcz, Poland
Received: 2006.01.10, Accepted: 2006.04.03, Published online first: 2006.10.06
Abstract
Introduction: Thalidomide is a derivative of glutamic acid with anti-angiogenic, anti-inflammatory, immunomodulatory and
anti-cancer properties that was found to inhibit the production of tumor necrosis factor α in vitro, stimulate reactive oxygen
species production, and inhibit vascular endothelial growth factor receptor in acute leukemias. The purpose of this study was
to determine the in vitro activity of thalidomide as a single agent and in combination with prednisolone or cytarabine in child-
hood acute lymphoblastic leukemia (ALL).
Materials and Methods: Bone marrow samples of 40 childhood ALL patients, normal lymphocytes of 9 healthy adults, and 3
lymphoid cell lines were evaluated for cytotoxicity of thalidomide (alone and in combination with prednisolone and cytara-
bine) using the MTT assay. Cell cycle analysis was performed by flow cytometry.
Results: Thalidomide as a single agent had weak antileukemic activity to the childhood ALL samples. However, in the pres-
ence of thalidomide the cytotoxicities of prednisolone and cytarabine were increased 3.3-fold (p<0.001) and 2.7-fold
(p=0.002), respectively. Thalidomide increased apoptosis in lymphoblasts and modulated the cell-cycle arrest caused by
prednisolone, but not that by cytarabine, in childhood ALL samples.
Conclusions: Thalidomide increases in vitro the sensitivity of childhood ALL cells to prednisolone and cytarabine.
Key words: thalidomide, prednisolone, cytarabine, drug resistance, acute lymphoblastic leukemia, children.
Regulation of cytokine transcription
in the context of chromatin
Barbara Nikolajczyk
Department of Microbiology, Boston University School of Medicine, Boston, MA 02118, USA
Received: 2006.04.12, Accepted: 2006.08.02, Published online first: 2006.10.06
Abstract
Understanding the transcriptional regulation of an important class of innate and adaptive immune system effector molecules,
the cytokines, is increasingly important given the promise cytokine regulation holds for treating various autoimmune and
inflammatory diseases. Studies defining the mechanisms regulating cytokine transcription initially focused on identifying the
cis-acting elements and trans-acting factors that activate cytokine promoters and enhancers. In the past, these studies were
largely completed in the absence of constraints instituted by cellular chromatin. Over the past decade it has become obvious
that changes in chromatin accessibility critically control, rather than simply correlate with, the transcriptional activation of
most genes, including cytokines. Hence candidate transcriptional activators are being re-evaluated for potency in the context
of cellular chromatin. Several distinct mechanisms for manipulating the generally repressive context of chromatin have been
identified for cytokine genes. Most recently, single nucleotide polymorphisms in cytokine transcriptional regulatory elements
have been shown to play measurable roles in regulating cytokine levels in the context of naturally selected haplotypes.
Overall, subtle differences in DNA sequence and nucleoprotein complex composition, including protein post-translational
modification, come together in cell type-specific combinations to explain the normal variation in cytokine transcription
throughout the human populace.
Key words: cytokine transcription, chromatin remodeling, haplotypes.
Mechanisms of tumor escape:
role of tumor microenvironment
in inducing apoptosis of cytolytic effector cells
Alessandro Poggi1 and Maria Raffaella Zocchi2
1 Laboratory of Experimental Oncology D, National Institute for Cancer Research, 16132 Genoa, Italy
2 Laboratory of Tumor Immunology, Scientific Institute San Raffaele, 20132 Milan, Italy
Received: 2006.06.02, Accepted: 2006.08.31, Published online first: 2006.10.06
Abstract
Spontaneous tumors grow and kill the host unless therapy reduces their mass to a level where the immune system, it is
thought, can control their growth and diffusion. Indeed, in many instances tumors can reappear, become resistant to thera-
py, and escape the host immune response. Many mechanisms of tumor escape operating in the tumor microenvironment
have been proposed: 1) low or absent expression of molecules on tumor cells involved in tumor target cell recognition; 2)
absence of co-stimulation leading to tolerization of T cells; 3) soluble factors secreted by tumor cells inhibiting T cell
response; and 4) regulatory T cells, myeloid suppressor cells, and stromal cells may impair immune-cell responses to tumors.
Furthermore, tumors can release soluble molecules such as HLA-I (sHLA-I). This, in turn, reduces T cell-mediated immune
response and induces apoptosis of cytolytic effector cells such as natural killer and CD8+ T lymphocytes through the engage-
ment of HLA-I receptors such as CD8 and/or activating isoforms of the inhibitory receptor superfamily. The release of sol-
uble ligand for activating receptors, e.g. UL16 binding proteins and/or MHC class I-related proteins A and B, the natural lig-
ands of NKG2D, may impair activation, effector cell-mediated recognition, and cytolysis of tumor cells. Furthermore, the
elimination of anti-tumor effector cells may be achieved by induction of apoptosis consequent to triggering elicited via acti-
vating molecules, such as receptors responsible for natural cytotoxicity, upon their binding with ligands expressed on tumor
cells.
Key words: tumor escape, soluble HLA-I, natural cytotoxicity receptor, NKG2D, apoptosis, stromal cells.
Langerhans cells in vulvar lichen sclerosus
and vulvar squamous cell carcinoma
Helena Rotsztejn1, Ewa Trznadel−Budźko2 and Dorota Jesionek−Kupnicka3
1 Section of Dermatology, Research Institute of the Polish Mother’s Memorial Hospital, £ódŸ, Poland
2 Department of Dermatology and Pediatric Dermatology, Military-Medical Faculty, Medical University of £ódŸ, Poland
3 Department of Tumor Pathology, Chair of Oncology, Medical University of £ódŸ, Poland
Received: 2006.02.27, Accepted: 2006.07.20, Published online first: 2006.10.06
Abstract
Introduction: Langerhans cells (LCs), specializing in antigen presentation, are a very important part of the skin immune sys-
tem (SIS).
Materials and Methods: Skin biopsies from 22 women with vulvar lichen sclerosus (LS): 15 patients with early and 7 with the
late stage of the disease, were evaluated. Five women with vulvar squamous cell carcinoma (SCC) were also examined. The
control group consisted of 9 women who underwent plastic surgery of the vulvar region. Immunohistochemical staining was
performed on formalin-fixed paraffin-embedded tissues samples using antihuman CD1a antibody (NCL-CD1a-235,
Novocastra).
Results: Increased numbers of LC stainings were present in early LS, whereas decreased numbers of these cells were present
in late LS and in SCC compared with the control group.
Conclusions: This study showed that dysregulation of the SIS may lead to suppression of LCs in the vulvar epithelium and
may be one of the reasons for a higher tendency for carcinogenesis in the vulvar region.
Key words: lichen sclerosus, vulva, Langerhans cells, squamous cell carcinoma
Increased whole blood chemiluminescence
in patients with chronic renal failure independent
of hemodialysis treatment
Jacek Rysz1, Elżbieta Potargowicz2, Maciej Banach3, Maria Łuczyńska2,
Robert Stolarek2, Piotr Białasiewicz2, Marek Kasielski4, Aleksandra Ciałkowska−Rysz5
and Dariusz Nowak2
1 Department of Internal Medicine and Dialysotherapy, Medical University of £ódŸ, Poland
2 Department of Experimental and Clinical Physiology, Institute of Physiology and Biochemistry,
Medical University of £ódŸ, Poland
3 Department of Cardiac Surgery, Medical University in £ódŸ, University Hospital No. 3, £ódŸ, Poland
4 Practical Clinical Training Center, Medical University of £ódŸ, Poland
5 Department of Palliative Care, Medical University of £ódŸ, Poland
Received: 2005.09.01, Accepted: 2006.03.22, Published online first: 2006.10.06
Abstract
Introduction: The luminol-enhanced whole blood chemiluminescence (LBCL) assay is a rapid assay for the measurement of
reactive oxygen species (ROS) generation by circulating phagocytes. This study’s aim was to determine if patients on main-
tenance hemodialysis (HD) and non-dialyzed patients with chronic renal failure (CRF) have altered LBCL and if dialysis
itself affects ROS production in the blood.
Materials and Methods: Twenty-six HD patients, 11 non-dialyzed patients with CRF, and 20 gender- and age-matched healthy
controls were studied. Resting (rCl) and 2×10–5 M n-formyl-methionyl-leucyl-phenylalanine-stimulated LBCL (peak chemi-
luminescence: pCl, total light emission after agonist addition: tCl) calculated per 104 phagocytes present in the 3-µl blood
samples were measured with a Bio-Orbit® 1251 luminometer at 37°C for 11 min.
Results: Prior to the HD session, median rCL, pCL, and tCL were 1.5, 3.0, and 2.8 times higher in HD patients than in healthy
controls (p<0.01) and tended to increase at the end of the session. Significant increases in tCl were observed at 30 min and
240 min (end) of HD (1023.5 vs. 1810.6 vs. 2006.8 arbitrary units×s/104 phagocytes, n=9, p<0.05). Median pCl and tCl were
5.0 and 4.3 times higher in non-dialyzed patients with CRF than in healthy controls (p<0.001). However, no significant differ-
ences were found between pre- and post-HD LBCL of HD patients and the LBCL of non-dialyzed patients with renal failure.
Conclusions: Blood from patients with renal failure generates elevated amounts of oxidants independently of HD treatment.
This may add to the understanding of the nature of oxidative stress and suggests the need of anti-oxidant treatment in these
patients.
Key words: uremia, hemodialysis, whole blood chemiluminescence, reactive oxygen species.
All roads lead to Rome:
pathways of NKT cells promoting asthma
Tan Jinquan, Wang Li, He Yuling and Chen Lang
Department of Immunology, and Laboratory of Allergy and Clinical Immunology, Institute of Allergy and Immune-related
Diseases and Center for Medical Research, Wuhan University School of Medicine, 430071 Wuhan, China
Received: 2006.06.29, Accepted: 2006.09.15, Published online first: 2006.10.06
Abstract
NKT cells are the prominent manipulator in asthma development. Asthmatic NKT cells migrate from thymus, spleen, liver
and bone marrow into blood vessels, and then concentrate in airway bronchi mucosa. This recruitment is dependent on high
expression of CCR9 and engagement of CCL25/CCR9. NKT cells promote asthma in two different pathways. One is an indi-
rect pathway. NKT cells contact with CD3+ T cells and induce them secreting large quantity of Th2 cytokines (IL-4, IL-13),
which requires the participation of dentritic cells and the synergic signaling of CCL25/CCR9 and CD226. The other is a
direct pathway. Circulating asthmatic NKT cells selectively highly express Th1 cytokines (IFN-γ). Once reached airway
epithelium, most NKT cells shift to Th2-bias, highly expressing IL-4, IL-13, but not IFN-γ. Both pathways lead to airway
hyperresponsiveness and inflammation, asthma development. Comparing to the well documented suppressive regulatory T
cells, CD4+CD25+ T cells, NKT cells perform as a novel active regulator in asthma. These recent understanding of NKT cells
performance in the development of asthma might unveil new therapy targets and management strategies for asthma.
Key words: NKT cells, asthma, chemokines, cellular signal transduction.
The effect of packed red blood cell storage
on arachidonic acid and advanced glycation
end-product formation
Lidia Łysenko1, Magdalena Mierzchała1, Andrzej Gamian2, Grażyna Durek1,
Andrzej Kübler1, Ryszard Kozłowski3 and Marek Śliwiński1
1 Department of Anesthesiology and Intensive Care, Wroc³aw University of Medicine, Wroc³aw, Poland
2 Department of Infectious Disease, Institute of Immunology and Experimental Therapy, Wroc³aw, Poland
3 Regional Center of Blood Donation and Blood Therapy, Wroc³aw, Poland
Received: 2005.11.02, Accepted: 2006.03.22, Published online first: 2006.10.06
Abstract
Introduction: The transfusion of packed red blood cells (PRBCs) is a significant risk to blood recipients. Blood banking pro-
cedures permit the storage of PRBCs for up to 42 days. Storage of PRBCs can cause polymorphonuclear granulocytes
(PMNs) activation and the development of neutrophil-mediated transfusion-related acute lung injury. The aim of our study
was to determine if PRBC storage has an influence on the formation of arachidonic acid (AA) and advanced glycation end
products (AGEs).
Materials and Methods: Twenty units of PRBCs were used to measure AA and AGE levels. The samples were taken on the
0th, 14th, 28th, and 42nd days of PRBC storage. The AA level was analyzed by gas-liquid chromatography-mass spectrome-
try and AGE level by an immunoenzymatic test.
Results: During the first 14 days of PRBC storage, the AA level significantly increased and then slowly decreased. The AGE
level increased continuously during the whole time of the study. In a model experiment, the AA glycoxidation product trans-
2-nonenal (T2N) formed adducts in reaction with hemoglobin which were detectable with the test for AGE.
Conclusions: It is highly probable that the observed increase in AGE level is related to the decrease in AA in PRBCs, which
can be associated with the formation of toxic aldehydes, especially T2N and 4-hydroxynonenal (HNE), from AA. Glucose in
the PRBCs (preservative solution) can contribute to AGE formation as well. The formation of AGEs, HNE, and T2N in
PRBCs, their influence on PMNs in vitro, and confirmation of our assumption need further studies.
Key words: packed red blood cells, polymorphonuclear granulocytes, arachidonic acid, 4-hydroxynonenal, trans-2-nonenal,
advanced glycation end products.
Modulation of the immune response
by extracellular matrix proteins
Sarah R. Morwood1 and Lindsay B. Nicholson1, 2
1 Department of Clinical Sciences South Bristol, School of Medical Sciences, University of Bristol, Bristol, UK
2 Department of Cellular and Molecular Medicine, School of Medical Sciences, University of Bristol, Bristol, UK
Received: 2006.06.19, Accepted: 2006.09.25, Published online first: 2006.11.21
Abstract
Inflammation entrains a focused and coordinated response from many different elements. Soluble factors such as
chemokines and cytokines direct the recruitment, differentiation, and fate of leukocytes. Cells and pathogens are killed and
consumed, yet where the response is effective, inflammation will melt away, leaving a healthy functioning tissue. All this com-
monly takes place in an environment known as the extracellular matrix (ECM). The ECM is not a passive partner in the
process and recent work demonstrates the important role that proteins found in this environment play in connecting differ-
ent parts of the immune response together. In this review we will focus on these connections and the proteins that make
them. One emerging trend that we will highlight is the ability of endogenous molecules to interact with receptors that are
better known as sensors of the molecular fingerprints of infection. We propose that this may be particularly relevant in the
context of autoimmunity, since the provision of such signals may be crucial in breaking tolerance.
Key words: extracellular matrix, inflammation, matricellular protein.
Hematopoietic cell transplantation
for chronic myeloproliferative disorders
William Tse1 and H. Joachim Deeg2
1 Division of Hematology/Oncology, Ireland Cancer Center, University Hospitals of Cleveland,
Case Western Reserve University School of Medicine, Cleveland, OH, USA
2 Clinical Research Division, Fred Hutchinson Cancer Research Center, and University of Washington, Seattle, WA, USA
Received: 2006.09.13, Accepted: 2006.10.02, Published online first: 2006.11.21
Abstract
Myeloproliferative disorders, including chronic idiopathic myelofibrosis (CIMF), polycythemia vera (PV), essential throm-
bocythemia (ET), and chronic myelomonocytic leukemia (CMML), are clonal diseases of hematopoietic stem or precursor
cells. They often show a protracted or chronic course; however, all have the potential of progressing to severe marrow fail-
ure, associated with myelofibrosis, or of transforming into acute leukemia. At that point, hematopoietic cell transplantation
(HCT) is the only current treatment strategy with curative potential. If transplantation is being considered and a suitable
donor is available, HCT should be carried out before leukemic transformation has occurred, as the success rate of HCT
declines steeply in patients who have evolved to leukemia. As many as 75–80% of patients with the original diagnoses of PV
or ET, about 65–70% with CIMF, and 45% of patients with CMML are surviving long term after allogeneic HCT using con-
ventional transplant regimens, with follow-up now extending to 15 years. Results with HLA-identical related and unrelated
donors are comparable. Major risk factors for the outcome after HCT are the disease stage, the presence of comorbid con-
ditions, and patient age. The development of reduced-intensity conditioning regimens has allowed for successful HCT even
for older patients and patients with comorbid conditions. Studies on disease mechanisms, including the recent characteriza-
tion of an activating mutation in JAK2, may provide additional prognostic guidance and are likely to lead to the develop-
ment of novel treatment strategies, which will require continuous reassessment as to the optimum timing of HCT.
Key words: hematopoietic cell transplantation, myeloproliferative disorders, myelofibrosis, polycythemia vera, essential throm-
bocythemia.
Control of immune responses
by immunoregulatory T cells
Geordie Rudge, Paul A. Gleeson and Ian R. van Driel
Department of Biochemistry and Molecular Biology, Bio21 Molecular Science and Biotechnology Institute,
The University of Melbourne, Parkville, Victoria 3010, Australia
Received: 2006.05.15, Accepted: 2006.07.25, Published online first: 2006.11.21
Abstract
Immunoregulatory T cells play a key role in modifying the immune responses to self antigens, tumor antigens, and patho-
genic organisms. This review summarizes recent data on naturally occurring CD4+ regulatory T cells that constitutively
express CD25 (CD25+ Treg). We examine the markers that can be used to differentiate these cells from effector T cells, what
is known about their mode of action in controlling the activity of effector T cells, the antigenic specificity of CD25+ Treg, and
their ability to survive and to be selected in vivo. We also summarize specific information on the role of CD25+ Treg in con-
trolling anti-tumor responses, an area were manipulation of this subset holds particular clinical promise.
Key words: regulatory T cells, tumor immunity, immunoregulation
Classification of
Proteus mirabilis TG 115
and CCUG 10701 into the
Proteus O23
serogroup based on chemical
and serological studies of O-polysaccharides
Agnieszka Zabłotni1, Andrei V. Perepelov2, Katarzyna Kołodziejska1, Krystyna Zych1,
Yuriy A. Knirel2 and Zygmunt Sidorczyk1
1 Department of General Microbiology, Institute of Microbiology and Immunology, University of £ódŸ, £ódŸ, Poland
2 N.D. Zelinsky Institute of Organic Chemistry, Russian Academy of Sciences, 119991 Moscow, Russian Federation
Received: 2005.12.15, Accepted: 2006.02.02, Published online first: 2006.11.21
Abstract
Introduction: Bacteria of the genus Proteus are a common cause of urinary tract infections. The O-polysaccharide (OPS) chain
of their lipopolysaccharide (LPS) defines the serological specificity of strains. Based on the OPS structures and the
immunospecificity of the LPS, Proteus strains have been classified into 74 O-serogroups.
Materials and Methods: The OPS of P. mirabilis TG 115 was obtained by mild acid degradation of the LPS and studied by
1H and 13C nuclear magnetic resonance spectroscopy. Antisera were raised by immunization of rabbits with heat-killed bac-
teria. Serological studies were performed using enzyme immunosorbent assay, passive immunoheamolysis, inhibition exper-
iments, absorption of O-antisera, and Western blot.
Results: The following structure of the P. mirabilis TG 115 OPS was established:
→2)-β-D-GalpA-(1→3)-α-D-GalpNAc-(1→4)-α-D-GalpA-(1→3)-β-D-GlcpNAc-(1→
The same structure has been reported previously for the O-polysaccharides of P. mirabilis CCUG 10701 (O74) and
P. mirabilis 41/57 (O23), except that they contain O-acetyl groups in non-stoichiometric quantities. Serological studies
showed the antigenic identity of the three strains and their close serological relatedness to P. vulgaris 44/57.
Conclusions: Based on the OPS structures and serological data, it is suggested to classify P. mirabilis 41/57, TG 115, and
CCUG 10701 into one subgroup and P. mirabilis 42/57 and P. vulgaris 43/57 and 44/57 into another subgroup of the Proteus
O23 serogroup.
Key words: Proteus, lipopolysaccharide, O-polysaccharide, O-serogroup, serological classification.
The effects of the initial treatment phase
and of adjunctive low-dose doxycycline therapy
on clinical parameters and MMP-8, MMP-9,
and TIMP-1 levels in the saliva and peripheral blood
of patients with chronic periodontitis
Renata Górska and Małgorzata Nędzi−Góra
Department of Periodontology and Oral Diseases, Institute of Dentistry, Medical University of Warsaw, Poland
Received: 2006.04.03, Accepted: 2006.09.29, Published online first: 2006.11.21
Abstract
Introduction: The treatment of periodontal disease can consist of bacterial plaque reduction, risk factor elimination, and met-
alloproteinase inhibitor medication. The level of matrix metalloproteinases (MMPs) are regulated by endogenous tissue
inhibitors of metalloproteinases (TIMPs) as well as therapeutic low-dose doxycycline. The aim of the study was to evaluate
the effect of the initial phase of periodontal treatment and the effect of doxycycline on clinical parameters and the MMP-8,
MMP-9, and TIMP-1 concentrations in the saliva and peripheral blood of patients with chronic periodontitis.
Materials and Methods: The study group consisted of 33 patients with chronic periodontitis. Conventional periodontal treat-
ment (scaling and root planing) was conducted on all the patients and doxycycline (20 mg orally) was administered twice daily
for three months. Thirty-three controls received the conventional treatment only. Clinical scores (PI, BI, PD, CAL) were
recorded before and three months after the treatment. MMP-8, MMP-9, and TIMP-1 concentrations in saliva and periph-
eral blood were measured by ELISA before and after the treatment of 20 patients from the study group and 13 of the con-
trols.
Results: The application of doxycycline 20 mg resulted in significant improvement in clinical parameters compared with the
conventional periodontal treatment. Doxycycline did not produce significant reductions in MMP-8 and MMP-9 levels in sali-
va observed after the conventional treatment. The study revealed increases in the TIMP-1 concentration and the MMP-
-8/TIMP-1 and MMP-9/TIMP-1 ratios in saliva and blood after treatment with doxycycline.
Conclusions: The study confirmed the modulating effect of doxycycline on the host response in chronic periodontitis.
Key words: chronic periodontitis, therapy, metalloproteinases
Anti-tumoral capabilities of effector cells after IFN-αα
or CpG-motif treatment of cocultured dendritic cells
Michael Erhardt1, 2*, Ingo G. H. Schmidt−Wolf1*, Elisabeth Sievers1, Susanne Frank1,
John Strehl1, Marie von Lilienfeld−Toal1 and Marcus Gorschlüter1
1 Department of Internal Medicine I, University of Bonn, Germany
2 Present address: North West Cancer Research Fund Institute, University of Wales, Deiniol Road, Bangor,
Gwynedd, LL57 2UW, UK
Received: 2006.05.17, Accepted: 2006.09.29, Published online first: 2006.11.21
Abstract
Introduction: Ex vivo expansion of monocyte-derived dendritic cells (mDCs) and subsequent coculture with autologous
cytokine-induced killer (CIK) cells is an established system to create specific and non-specific anti-tumoral immunity. mDCs
constitute the most frequently applied DC subset in clinical studies. One recently published approach to optimize the
immunological functions of the DC/CIK cell system is the replacement of interleukin (IL)-4 by interferon (IFN)-α in the
maturation process of the DCs.
Materials and Methods: The expressions of relevant surface antigens of IL-4-DCs and IFNα-DCs by flow cytometry and the
anti-tumoral activation of effector cells cocultured with both types of DCs using cytotoxicity assays were compared. In addi-
tion, short-term coculture experiments with both types of DCs and IFNγ-LAK effector cells were performed and compared
with standard CIK cell coculture experiments.
Results: Regarding the expressions of functionally relevant surface markers, no differences could be detected for CD80,
CD83, and HLA-DR between IFNα-DCs and IL-4-DCs, whereas the mean fluorescence intensities of CD40, CD86, CD54,
and HLA-ABC were decreased and the expression of CD14 was increased for IFNα-DCs. Moreover, no enhancement of
cytotoxicity of cocultured CIK cells against tumor cell lines (A498 and SW480) was detected by the use of IFNα-DCs.
Additionally, coculture experiments with IFNγ-LAK cells were performed and unexpectedly higher lysis rates in comparison
with the established IL-4-DC/CIK coculture model was observed. Early incubation of the mDCs with several CpG-ODNs
failed to increase the anti-tumoral cytotoxicity of the cocultured IFNγ-LAK cells.
Conclusions: These results demonstrate that in the mDC/CIK cell system, IFNα-DCs are not superior in inducing anti-
-tumoral cytotoxicity and even moderately inferior regarding the expression of functionally relevant surface markers com-
pared with IL-4-DCs.
Key words: dendritic cells, CIK cells, LAK cells, interferon-α, interleukin-4, CpG-ODN
Intracellular signaling pathways
in IgE-dependent mast cell activation
Agnieszka Kopeć, Bernard Panaszek and Andrzej M. Fal
Department of Internal Medicine and Allergology, Wroc³aw Medical University, Poland
Received: 2005.12.14, Accepted: 2006.08.07, Published online first: 2006.11.21
Abstract
Mast cells (MCs) are both central effectors and signaling cells in allergic reactions. Their key role in the immunopathology
of asthma and other allergic diseases has been well documented. Molecular events leading to MC activation have not been
yet fully established, however. Recent studies emphasize the key role of the protein tyrosine kinases Lyn and Fyn in MC sig-
nal transduction. The finding that Lyn kinase negatively regulates MC degranulation and that Fyn kinase enhances this effec-
tor response is of great importance. This creates new possibilities for therapeutic intervention in asthma and other allergic
diseases. This review summarizes current knowledge on MC intracellular signaling and discusses the most recent strategies
for the treatment of allergic diseases based on MC signaling pathway inhibition.
Key words: mast cell activation, intracellular signaling, FcεRI, Fyn kinase, Lyn kinase, phospholipase-C, MAPK.
ESH Euroconference on Translational Research
in Transplantation: immunogenetics,
pharmacogenomics, proteomics, and immunobiology
Grzegorz Władysław Basak, Anna Gronkowska