Eicosanoid regulation of pulmonary innate immunity post-hematopoietic stem cell transplantation
Megan N. Ballinger, Tracy R. McMillan and Bethany B. Moore
Abstract
Hematopoietic stem cell transplantation (HSCT) is a therapeutic option for a number of malignant and inherited disorders. However, the efficacy of this therapy is limited by a number of serious infectious and noninfectious complications. Pulmonary infections represent a significant cause of morbidity and mortality post-HSCT and can occur both pre- and post-hematopoietic reconstitution. Susceptibility to Gram-negative bacterial infections despite full hematopoietic engraftment suggests that innate immunity remains impaired months to years post-HSCT. This review will describe the process and complications of HSCT and will summarize what is known about innate immune reconstitution post-HSCT. Data from the literature as well as our own laboratory will be presented to suggest that an eicosanoid imbalance characterized by over-production of prostaglandins and under-production of leukotrienes leads to impaired lung phagocyte function post-HSCT. Of therapeutic interest, strategies which limit production of prostaglandins can improve pulmonary host defense in animal HSCT models, which suggests that this may also be beneficial for human HSCT recipients.
Key words: transplantation, macrophages, neutrophils, innate immunity, eicosanoids, prostaglandins, leukotrienes.
Treatment of autoimmune disease: time for a paradigm shift?
Felix Mor
Abstract
Current treatment of human autoimmune diseases (AIDs) was developed empirically and relies mostly on non-selective suppression of the immune system. Traditional non-selective immunosuppressants such as corticosteroids, cyclophosphamide, and methotrexate and more novel means such as monoclonal antibodies to CD3, CD4, or CD25 do not discriminate between pathogenic and beneficial T cells. Importantly, the severe side effects seen with current therapies are related to the fact that these treatments not only suppress the pathogenic disease-inducing cells, but also cells influential in combating infections and killing malignant cells. Severe infections and malignancies are the inevitable result of non-selective immune suppression. Many of the novel forms of therapy of AID were developed in experimental animals, and their translation to the human disease was associated with the revelation of unexpected and sometimes catastrophic side effects. These surprises underscore the major differences between the relative simplicity of the experimental model and the complexity of the human disease. How can this current state of treatment of AID be improved? Which principles should guide us in the design of new treatments? This review attempts to offer a new look at these questions.
Key words: autoimmune disease, immune regulation, immune suppression, regulatory T cells.
Expression of interleukin-16 by tumor-associated
macrophages/activated microglia in high-grade
astrocytic brain tumors
Markus Liebrich1*, Liang−Hao Guo1*, Hermann J. Schluesener1, Jan M. Schwab1, 4#,
Klaus Dietz2, Bernd E. Will3 and Richard Meyermann1
1 Institute of Brain Research, University of Tuebingen, Medical School, Tuebingen, Germany
2 Department of Medical Biometry, University of Tuebingen, Medical School, Tuebingen, Germany
3 Department of Neurosurgery, University of Tuebingen, Medical School, Tuebingen, Germany
4 CNRS UMR 7102 Université Pierre et Marie Curie, Equipe Developpment Neuronal, 9 Quai Saint Bernard, 75005,
Paris, France
# Present address: Brigham and Women’s Hospital, Center for Experimental Therapeutics, 75 Francis Street, Thorn
Building for Medical Research 724, Harvard Medical School, Boston, MA 02115, USA
Received: 2006.04.03, Accepted: 2006.10.06, Published online first: 2007.01.15
Abstract
Introduction: Macrophages/microglial cells are considered as immune cells in the central nervous system. Interleukin (IL)-16
is a proinflammatory cytokine produced by activated monocytic cells.
Materials and Methods: Expression of IL-16 was analyzed by immunohistochemistry in human astrocytic brain tumors and the
rat C6 glioblastoma tumor model. IL-16 was detected in both human astrocytic brain tumors and rat C6 glioma.
Results: Compared with human control brains, a significant increase in the percentages of parenchymal IL-16+
macrophages/microglia was observed already in grade II astrocytomas, indicating that IL-16+ immunostaining could be
a descriptor of a macrophage/microglia subset in astrocytic brain tumors. A further increase was observed at the transition
from grade II to III astrocytomas. This increase in IL-16 immunoreactivity correlated with WHO grades of human astrocyt-
ic brain tumors.
Conclusions: Therefore, IL-16 might be a so far unknown factor in the regulation of the local inflammatory milieu of human
and experimental astrocytomas.
Key words: interleukin-16, microglia, astrocytic brain tumor, inflammation, immunohistochemistry.
Immunological system status
and the appearance of respiratory system disturbances
in thymectomized patients
Paweł Krawczyk1, Marta Adamczyk−Korbel1, Robert Kieszko1, Elżbieta Korobowicz2
and Janusz Milanowski1
1 Department of Pneumonology, Oncology, and Allergology, Medical University of Lublin, Poland
2 Department of Pathomorphology, Medical University of Lublin, Poland
Received: 2006.01.02, Accepted: 2006.10.06, Published online first: 2007.02.02
Abstract
Introduction: Adult-onset thymoma may be responsible for several diseases, such as pure red cell aplasia, myasthenia gravis,
and immunodeficiency (Good’s syndrome). Thymectomy does not always improve the patient’s condition, and may even pro-
duce additional symptoms. Its pathogenesis is still not entirely understood, but autoimmunological processes and bone mar-
row defect are the most frequently suggested.
Materials and Methods: Eleven patients (mean age: 56.2±15.5 years) were analyzed 6 months to 10 years after thymectomy
due to thymoma as were 25 healthy persons serving as controls. Enzyme-linked immunosorbent assay (ELISA) and flow
cytometry techniques were used to evaluate the immunological status of the subjects.
Results: Good’s syndrome was diagnosed in one patient, 4 subjects suffered from myasthenia gravis, and recurrent infections
of upper and lower respiratory tract appeared in 9 patients. The immunological analyses (ELISA and flow cytometry)
revealed a significantly lower IgG level (p<0.05), percentage of peripheral blood B lymphocytes (p<0.0005), and CD4:CD8
ratio (p<0.05) in thymectomized patients compared with the healthy controls. The percentages of CD4+ and CD8+ T lym-
phocytes expressing CD28 antigen were significantly lower in thymectomized patients than in healthy subjects (p<0.005 and
p<0.01, respectively). The percentage of naïve T helper lymphocytes was significantly lower in the patients than in the control
group (p<0.05).
Conclusions: Immunodeficiency and recurrent infections may be the first symptoms of immunological disturbances after
thymectomy in adults. It is suggested that regular medical monitoring of these patients is important in preventing further
complications, which may result in irreversible lung tissue destruction.
Key words: thymectomy, immunodeficiency, respiratory infections.
Peroxisome proliferator-activated receptor γγ
(PPARγγ) and sepsis
Andreas von Knethen, Mathias Soller and Bernhard Brüne
Department of Biochemistry I-Pathobiochemistry, Johann Wolfgang Goethe-University Frankfurt, Faculty of Medicine,
60590 Frankfurt am Main, Germany
Received: 2006.10.04, Accepted: 2006.12.21, Published online first: 2007.02.02
Abstract
This review describes the role of the nuclear hormone receptor PPARγ as a double-edged sword in sepsis. On the one hand,
PPARγ inhibits pro-inflammatory gene expression, predominantly by scavenging transcription factors and their cofactors,
thus preventing them from binding to their cognate binding sites in the promoters of target genes. The expressions of the
affected genes, such as those for inducible nitric oxide synthase, TNF-α, or IL-1β, are repressed. Therefore, PPARγ is sug-
gested to be beneficial in hyper-inflammatory diseases, such as sepsis. In animal models of sepsis, PPARγ agonist pretreat-
ment auspiciously attenuated inflammation compared with control animals, accompanied by their improved survival rate. On
the other hand, PPARγ provokes apoptosis, which in the hyper-inflammatory phase of sepsis might be helpful because the
number of immune cells, such as monocytes, macrophages, and neutrophils, involved in secreting high amounts of pro-
inflammatory mediators will be reduced. In contrast, during the anti-inflammatory phase, cell death of immune cells, espe-
cially of T lymphocytes, is supposed to be deleterious. Under these circumstances, a second infection cannot be adequately
answered, thus causing septic shock and multi-organ dysfunction syndrome. Therefore the role of PPARγ is still ambiguous.
Particularly its role in initiating apoptosis awaits further clarification to finally elucidate its impact on sepsis development.
Key words: inflammation, T cells, sepsis, apoptosis.
Antichlamydial antibodies and citric acid
in patients with chronic prostatitis
Iwona Ostaszewska−Puchalska1, Bożena Zdrodowska−Stefanow2,
Jerzy Badyda3 and Zofia Galewska4
1 Center for Sexually Transmitted Disease Research and Diagnostics, Bia³ystok, Poland
2 Department of Dermatology and Venerology, Medical University, Bia³ystok, Poland
3 Urological Outpatient Clinic “Multimedica”, Bia³ystok, Poland
4 Department of Medical Biochemistry, Medical University, Bia³ystok, Poland
Received: 2006.05.04, Accepted: 2006.10.06, Published online first: 2007.02.02
Abstract
Introduction: The aim of the study was to evaluate the correlation between the presence of anti-C. trachomatis (C.t.) anti-
bodies in serum and expressed prostatic secretions (EPS) and the concentration of citric acid in patients with chronic pro-
statitis.
Materials and Methods: The study involved 34 men with chronic prostatitis. The leukocyte count, presence of anti-C.t. anti-
bodies (IgA, IgG), and citric acid concentration were determined in the EPS. The serum was examined for IgM, IgA, and
IgG anti-C.t. antibodies. Specific antibodies were determined using the EIA method. The concentration of citric acid was
measured using the ultraviolet method.
Results: Inflammation of the prostate (≥10 PMN) was found in 61.8% of the patients. A reduction in citric acid concentra-
tion in the EPS was detected in 58.8% of the men. Specific serum antibodies were detected in 58.8% of the patients, includ-
ing 23.5% with IgM, 32.4% with IgA, and 44.1% with IgG. In all patients, serum IgM and IgA antibody titers were low, while
those of IgG antibodies were strongly positive in 46.7% of the patients. Anti-C.t. antibodies in the EPS were detected in
44.1% of the patients, including 32.4% with IgA and 35.3% with IgG. In contrast to serum, the titers of IgG antibodies in
the EPS were low in all the patients, while those of IgA were strongly positive in 54.5% of cases. In patients with positive
serological outcomes, 85% had reduced concentrations of citric acid.
Conclusions: The occurrence of anti-C.t. antibodies is usually accompanied by a decrease in the concentration of citric acid
in the prostatic secretion.
Key words: Chlamydia trachomatis, citric acid, prostate gland, expressed prostatic secretions, antichlamydial antibodies.
Transcriptional regulation in thymic epithelial cells
for the establishment of self tolerance
Mitsuru Matsumoto
Division of Molecular Immunology, Institute for Enzyme Research, University of Tokushima, Tokushima, Japan
Received: 2006.10.10, Accepted: 2006.11.30, Published online first: 2007.02.02
Abstract
Thymic epithelial cells (TECs) play pivotal roles in the establishment of self tolerance through critical dialogue with devel-
oping thymocytes. Unique actions of two transcriptional regulators within TECs, NF-κB-inducing kinase (NIK) and an
autoimmune regulator (AIRE), for the establishment of self tolerance have recently been highlighted by studies using
a strain of mouse bearing a natural mutation of the NIK gene (aly mice) and gene-targeted mice, respectively. Previous stud-
ies have demonstrated essential roles of NIK downstream of the lymphotoxin-β receptor (LTβR), which is essential for the
development of secondary lymphoid organs; aly mice lack all lymph nodes and Peyer’s patches because of the defective LTβR
signaling. Additional roles of NIK in thymic organogenesis downstream of LTβR, mainly through the developmental regu-
lation of TECs, have now emerged, although the corresponding ligand(s) for LTβR participating in this action have not been
fully characterized. In contrast, AIRE, a gene responsible for the development of an organ-specific autoimmune disease that
demonstrates monogenic autosomal recessive inheritance, contributes to the establishment of self tolerance probably by con-
trolling the expression of self antigens through yet undetermined molecular mechanisms. Thus, it is highly likely that a group
of genes control self tolerance within TECs through unique and coordinated actions, and that an understanding of this
process would help to unravel the pathogenesis of autoimmune disease.
Key words: autoimmune disease, thymic epithelial cell, NF-κB-inducing kinase, aly mice, lymphotoxin, AIRE.
The use of magnesium in bronchial asthma:
a new approach to an old problem
Aneta Kowal, Bernard Panaszek, Wojciech Barg and Andrzej Obojski
Department of Internal Medicine and Allergology, Wroc³aw Medical University, Wroc³aw, Poland
Received: 2006.03.23, Accepted: 2006.11.24, Published online first: 2007.02.02
Abstract
Magnesium deficiency is a common electrolyte disorder in patients with acute severe asthma, but intracellular magnesium
content better reflects its homeostasis than does its serum concentration. Magnesium takes part in many metabolic process-
es in the organism, including energy metabolism, protein and nucleic acid synthesis, cell cycle, the binding of substances to
the plasma membrane, and maintenance of cytoskeletal and mitochondrial integrity. It also modulates ion transport and
influences intracellular calcium concentration. Maintenance of the cells’ transmembrane gradient depends on the presence
of magnesium, and hypomagnesemia may result in an increase in neuromuscular cell excitability. Magnesium is a cation mod-
ulating the smooth muscle contractility of different tissues: hypomagnesemia causes their contraction and hypermagnesemia
their relaxation. Suggestions of a positive influence of magnesium in the treatment of asthma exacerbation have been known
for a long time, but research results differ. A single dose of intravenous magnesium sulfate given to patients with acute asth-
ma exacerbation has been shown to be safe, but its efficiency is still under discussion. According to the Global Initiative for
Asthma GINA-2005, magnesium sulfate administration is not recommended for routine treatment, but it is permitted in
patients with severe asthma exacerbation not responding to treatment (evidence category A). Recommendations of the
British Thoracic Society allow one dose of magnesium sulfate to patients with acute severe asthma exacerbation and inade-
quate initial response to broncho-dilating inhalation treatment (evidence category A). Future investigations should help to
establish the indications for magnesium use in the treatment of acute asthma exacerbations as well as the magnesium dose
and the scheme of its administration.
Key words: magnesium, hypomagnesemia, smooth muscle contractility, acute asthma exacerbation.
Rho kinase in the regulation of cell death and survival
Jianjian Shi and Lei Wei
Division of Pediatric Cardiology, Department of Pediatrics, Herman B Wells Center for Pediatric Research,
Indiana University, School of Medicine, Indianapolis, IN, USA
Received: 2006.11.07, Accepted: 2006.12.21, Published online first: 2007.03.09
Abstract
Rho kinase (ROCK) belongs to a family of serine/threonine kinases that are activated via interaction with Rho GTPases.
ROCK is involved in a wide range of fundamental cellular functions, such as contraction, adhesion, migration, and prolifer-
ation. Recent studies have shown that ROCK plays an important role in the regulation of apoptosis in various cell types and
animal disease models. Two ROCK isoforms, ROCK1 and ROCK2, are assumed to be function redundant, this based large-
ly on kinase construct overexpression and chemical inhibitors (Y27632 and fasudil) which inhibit both ROCK1 and ROCK2.
Gene targeting and RNA interference approaches allow further dissection of distinct cellular, physiological, and patho-phy-
siological functions of the two ROCK isoforms. This review, based on recent molecular, cellular, and animal studies, focus-
es on the current understanding of ROCK signaling in the regulation of apoptosis and highlights new findings from recent-
ly generated ROCK-deficient mice.
Key words: ROCK1, ROCK2, apoptosis, survival, gene targeting.
Ligand-independent activity of the B cell
antigen receptor in physiology and pathology
Daniel Corcos
The Babraham Institute, Babraham, Cambridge, UK
Received: 2006.11.02, Accepted: 2006.12.21, Published online first: 2007.03.09
Abstract
The B cell receptor (BCR) is required for stimulation of B cells by antigen, and is also involved in the negative selection of
autoreactive B cells. In the past few years, a constitutive ligand-independent signaling activity of the BCR has been demon-
strated. In this paper, the various findings are summarized and their interpretation and their significance, both in pathology
and in physiology discussed. The constitutive activity of the BCR may be important for tumor formation, at least in the case
of heavy-chain diseases, neoplastic proliferations developed from B cells. A large body of evidence suggests that this activi-
ty could be required for B cell survival and would play a role in B cell development as a process monitoring BCR function-
ality. A model explaining signaling in the absence of antigen as a function of dimer formation is proposed. The putative con-
stitutive activity of the pre-BCR is also discussed.
Key words: B cell receptor, signaling, oncogenic growth factors, heavy-chain diseases, B cell development.
Regulation of autoreactive B cells:
checkpoints and activation
Chuanlin Ding and Jun Yan
Tumor Immunobiology Program, Department of Medicine, James Graham Brown Cancer Center, University of Louisville,
Louisville, KY 40202, USA
Received: 2006.08.18, Accepted: 2006.12.21, Published online first: 2007.03.09
Abstract
It has become clear that the autoreactive B cells are a part of the normal naïve B cell repertoire in the periphery, despite the
fact that they undergo a series of checkpoints, which include receptor editing (revision), clonal deletion, and anergy.
However, most of those B cells reactive against self antigen remain functionally naïve for autoantibody production by dif-
ferential peripheral checkpoints. Therefore, the presence of autoreactive B cells does not always signify disease. Regulation
of their activation and effector functions will determine the ultimate outcome. Although autoreactive B cell tolerance is well
maintained in the healthy individual, the existence of pathogenic autoantibodies in autoimmune diseases indicates that these
tolerogenic checkpoints are broken. Recent studies have demonstrated that autoreactive B cells are regulated by a compos-
ite of factors, such as genetic susceptibility and environmental triggers such as bacterial and viral infections as well as other
immune cells. Interestingly, Toll-like receptors, previously considered as pattern-recognition receptors to detect and sense
pathogens, may also have a potential to recognize self antigens and regulate autoreactive B cells for activation.
Understanding the mechanisms of autoreactive B cell regulation and activation may help in identifying novel targets for the
treatment of autoimmune diseases.
Key words: autoreactive B cells, tolerance, Toll-like receptors, infection.
Potential immunological functions of salivary Hsp70
in mucosal and periodontal defense mechanisms
Tibor Károly Fábián1, Pál Fejérdy1, Minh Tú Nguyen2, Csaba Sőti2 and Péter Csermely2
1 Clinic of Prosthetic Dentistry, Faculty of Dentistry, Semmelweis University Budapest, Budapest, Hungary
2 Department of Medical Chemistry, Faculty of Medicine, Semmelweis University Budapest, Budapest, Hungary
Received: 2006.06.29, Accepted: 2006.12.29, Published online first: 2007.03.09
Abstract
Molecular chaperones were considered to be intracellular, but there is increasing evidence demonstrating their cytoprotec-
tive and immune modulator properties outside the cell. The major extracellular chaperone (Hsp70) was also found in saliva,
indicating a possible effect of Hsp70 on mucosal surfaces. Here we summarize the immune-modulatory role of the 70-kDa
stress protein family, with special attention on the potential impact of salivary Hsp70 on oral defense mechanisms. There are
three major facets of Hsp70-induced immune activation: 1) the appearance of Hsp70 on the surface of certain tumor cells or
virally infected cells, leading to their phagocytosis and subsequent lysis; 2) the role of extracellular uncomplexed Hsp70 as
a danger signal, leading to the secretion of proinflammatory cytokines from antigen-presenting cells and T lymphocytes and
of nitric oxide from macrophages as well as to complement activation; 3) receptor-mediated uptake of peptide-loaded Hsp70
to antigen-presenting cells and cross-presentation of the Hsp70-peptide complex as an antigen to cytotoxic T cells and nat-
ural killer lymphocytes. The immune-activating effect of salivary Hsp70 may also be highly important in oral defense, espe-
cially in areas where molecular and cellular participants of the immune response appear on the surface of the oral cavity (i.e.
several lesions of the mucosa and the periodontal tissues).
Key words: Hsp70, extracellular, saliva, immune, mucosal, periodontal.
Cytokine production by human leukocytes
with different expressions of natural antiviral immunity
and the effect of antibodies against interferons
and TNF-αα
Beata Orzechowska, Zenon Antoszków, Iwona Siemieniec, Maria Lorenc,
Bogna Jatczak and Zofia Błach−Olszewska
Institute of Immunology and Experimental Therapy, Polish Academy of Sciences, Wroc³aw, Poland
Received: 2006.09.08, Accepted: 2006.11.27, Published online first: 2007.03.20
Abstract
Introduction: Two activities of innate antiviral immunity were studied: the resistance of human peripheral blood mononuclear
cells (PMBCs) ex vivo to viral infection and the production of cytokines.
Materials and Methods: Samples of blood were taken from healthy blood donors and from persons with frequent infections
of the upper respiratory system. PMBCs were isolated by gradient centrifugation. Vesicular stomatitis virus (VSV) was used
as the indicatory virus to infect PMBCs. The cytokines: IFN, TNF, and IL-6 were titrated by biological methods and IL-10
by ELISA.
Results: Blood donors were divided for two groups: those with VSV-resistant and those with VSV-sensitive PMBCs and
secretion of cytokines by them was compared. The resistant PMBCs produced more cytokines than the sensitive ones. A sta-
tistically significant difference, was found only in the case of the IFNs. To examine the contribution of IFNs and TNF in
maintaining resistance, leukocytes from both groups were treated with specific anti-cytokine antibodies. The authors’ previ-
ous study showed that the elimination of spontaneous IFN-α, IFN-β, IFN-γ, and TNF-α from resistant leukocytes resulted
in increased VSV replication This indicates the important role of cytokines. In VSV-sensitive PMBCs, anti-IFN-α showed
the opposite effect (decreased virus replication). In the absence of spontaneous IFN-α, disturbances in cytokine production
were observed.
Conclusions: Complete resistance of PMBC to VSV infection is accompanied by higher cytokine release, The paradoxical
effect of anti-IFN-α on virus replication in leukocytes sensitive to viral infection may be attributed to changes in the cytokine
profile balance, i.e. high TNF production by VSV-infected leukocytes and a complete reduction of IL-6 production.
Key words: cytokines, leukocytes, innate immunity, viral infection.
Regulation of the NADPH oxidase activity
and anti-microbial function of neutrophils
by arachidonic acid
Charles S. Hii1, 2 and Antonio Ferrante1, 2, 3
1 Department of Immunopathology, Children, Youth and Women’s Health Services, North Adelaide, South Australia
2 Department of Paediatrics, University of Adelaide, South Australia
3 School of Pharmacy and Medical Sciences, University of South Australia, Adelaide, Australia
Received: 2006.11.13, Accepted: 2007.01.31, Published online first: 2007.03.20
Abstract
Arachidonic acid (AA), a second-messenger molecule released from membrane phospholipids by phospholipase A2 in acti-
vated cells, is a stimulator of neutrophil responses, including the oxygen-dependent respiratory burst. The polyunsaturated
fatty acid is also the precursor of biologically active eicosanoids. There are several mechanisms by which AA stimulates the
respiratory burst. These include the direct binding of AA to S100 proteins which regulate the assembly of the NADPH oxi-
dase as well as the activation of key signaling molecules which control the respiratory burst. Arachidonic acid also stimulates
it own release from membrane phospholipids and this contributes to optimal respiratory burst activity. Thus, increased lev-
els of AA at sites of inflammation will influence the magnitude and course of the inflammatory response, not only by direct-
ly affecting the function of infiltrating neutrophils and other leukocytes, but also through its metabolites generated by lipoxy-
genases and cyclooxygenases.
Key words: neutrophils, arachidonic acid, anti-microbial function, respiratory burst, mechanism of action.
Glucose transport in human peripheral blood lymphocytes
influenced by type 2 diabetes mellitus
Paweł Piątkiewicz, Anna Czech and Jan Tatoń
Chair and Department of Internal Medicine and Diabetology, Medical University of Warsaw, Warsaw, Poland
Received: 2006.06.01, Accepted: 2006.12.28, Published online first: 2007.03.20
Abstract
Introduction: The objective of this study was to evaluate glucose transport into lymphocytes in healthy subjects and patients
with type 2 diabetes mellitus (DM) treated either with diet only or with insulin and to propose peripheral blood lymphocytes
as a convenient model for cellular glucose transport studies.
Materials and Methods: Sixty subjects with type 2 DM, 30 treated with diet only and 30 with insulin, were investigated. Thirty
healthy subjects matched for age, weight, and sex served as a control group. Deoxy-D-glucose, 2-[3H(G)] transport was stud-
ied in isolated peripheral blood lymphocytes. Expression of glucose transporters was ascertained by immunocytochemical
identification and by Western blotting.
Results: In lymphocytes from the control group, deoxy-D-glucose uptake increased gradually with the duration of the exper-
iment. In diabetics treated with insulin, the maximal increase in deoxy-D-glucose uptake was observed after 30 min of the
investigation, followed by a plateau phase. In diabetics treated with diet, deoxy-D-glucose uptake increased slowly during the
first 30 min. The presence of GLUT1 and GLUT3 in lymphocytes was confirmed in this study.
Conclusions: Glucose transport into lymphocytes is altered in type 2 DM. In lymphocytes from diabetics, the dynamics of
deoxy-D-glucose uptake significantly differed from that in healthy subjects. There was also a significant difference between
the diabetic groups, representing different modes of therapy and stages of the disease. Glucose transport into lymphocytes
is apparently influenced by DM as well as by the mode of therapy. We suggest that peripheral blood lymphocytes may
become a promising model for studies on glucose transport in diabetes.
Key words: type 2 diabetes mellitus, glucose transport, lymphocytes, GLUT.
Do Langerhans cells play a role
in vulvar epithelium resistance
to 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: 2006.04.27, Accepted: 2006.07.17, Published online first: 2007.03.20
Abstract
Introduction: Langerhans cells (LCs) are a very important part of the skin immune system.
Materials and Methods: Skin biopsies taken from 13 women after the removal of vulvar squamous cell carcinoma (SCC) who
had not been treated earlier for any vulvar diseases were investigated. The control group consisted of 12 women who under-
went a plastic surgical operation of the vulva region. Immunohistochemical staining was performed on formalin-fixed, paraf-
fin-embedded tissues samples using antihuman CD1a antibody (NCL-CD1a-235, Novocastra).
Results: This study showed a large decrease in LCs in vulvar SCC.
Conclusions: It is postulated that the reduction in the number of LCs may be one of the reasons for a higher tendency of car-
cinogenesis in the vulvar region. Their role as a main element of the skin immune system in the initiation of this process
needs further investigation. It is possible that research on LCs in the skin will cast a new light on their role and even con-
tribute to the prophylaxis and treatment of skin and mucosa carcinomas.
Key words: vulva, Langerhans cells, squamous cell carcinoma.
Vascular endothelial growth factor
in systemic lupus erythematosus: relationship
to disease activity, systemic organ manifestation,
and nailfold capillaroscopic abnormalities
Anna Kuryliszyn−Moskal, Piotr Adrian Klimiuk, Stanisław Sierakowski
and Mariusz Ciołkiewicz
Department of Rheumatology and Internal Diseases, Medical University of Bia³ystok, Bia³ystok, Poland
Received: 2006.10.18, Accepted: 2006.12.28, Published online first: 2007.06.08
Abstract
Introduction: The aim of the study was to evaluate whether vascular endothelial growth factor (VEGF) serum level is associ-
ated with systemic organ involvement, microvascular changes as determined by nailfold capillaroscopy, and disease activity
of systemic lupus erythematosus (SLE).
Materials and Methods: Serum levels of VEGF were determined by an enzyme-linked immunosorbent assay in 47 SLE
patients and in 30 healthy controls. Nailfold capillaroscopy was performed in all patients and healthy subjects.
Results: Morphological changes were observed by nailfold capillaroscopy in 45 of 47 (95.7%) SLE patients. Mild capillary
changes were found in 16 (34%), moderate in 21 (44.7%), and severe in 8 (17%) SLE patients. All patients with systemic
organ involvement showed severe or moderate changes in nailfold capillaroscopy. In comparison with the control group,
a higher serum concentration of VEGF in SLE patients was demonstrated (p<0.05). Furthermore, significant differences in
VEGF serum concentration between SLE patients with systemic involvement and controls were found (p<0.01).
Comparison between patients with active and inactive SLE according to SLEDAI score showed a significantly higher con-
centration of VEGF in the sera of patients with active SLE (p<0.01). The SLE patients with severe and moderate changes
in nailfold capillaroscopy showed significantly higher VEGF serum levels than SLE patients with mild changes (p<0.05) or
healthy controls (p<0.01). Moreover, the VEGF serum level correlated significantly with ESR (r=0.580, p<0.0001) and
CRP (r=0.512, p<0.005).
Conclusions: Our data suggest that VEGF serum level may be a useful marker of disease activity and internal organ involve-
ment in SLE patients. Abnormalities in nailfold capillaroscopy may reflect the extent of microvascular involvement and are
associated with systemic manifestation in SLE.
Key words: systemic lupus erythematosus, vascular endothelial growth factor, nailfold capillaroscopy, systemic involvement.
Effect of structural modification at the 4, 3’,
and 2’ positions of doxorubicin
on topoisomerase II poisoning, apoptosis,
and cytotoxicity in human melanoma cells
Beata M. Gruber1, Elżbieta L. Anuszewska1, Irena Bubko1, Aneta Goździk1,
Izabela Fokt2 and Waldemar Priebe2
1 Department of Biochemistry and Biopharmaceuticals, National Institute of Medicines, Warsaw, Poland
2 Department of Experimental Therapeutics, M.D. Anderson Cancer Center, Houston, TX 77030, USA
Received: 2007.01.23, Accepted: 2007.02.28, Published online first: 2007.06.08
Abstract
Introduction: The mechanism of the cytotoxicity of anthracyclines is pleiotropic and its significance in cell growth inhibition
seems to be highly specific and dependent on cell type and anthracycline drug. Resistance and the high cardiotoxicity of
anthracyclines have stimulated many studies aimed at identifying critical substituents required for optimal activity. Many
authors point to the fact that the double-strand breaks, the consequence of the activity of topoisomerase II poisons, and the
inability of cells to repair the DNA lesions are the signal for apoptosis. The aim of this study was to define the influence of
4-demetoxy 2’-halogenated analogs with altered basicity at the 3’-position on topoisomerase II and the relationship of that
interaction with apoptosis and the cytotoxicity of these novel anthracyclines. Parental human ME18 melanoma cells and the
ME18/R subline, obtained experimentally, resistant to doxorubicin (DOX), exposed to 1.7 and 8.6 µM DOX or its analogs,
annamycin and WP903 (both 0.3 and 3.0 µM) were studied.
Materials and Methods: The MTT test was used to assay cytotoxicity. Interaction of the drugs with topoisomerase II and apop-
tosis were done by Western blot and fluorescence microscopy using Hoechst 33342.
Results: The structural changes at positions 4, 2’, and 3’ can influence topoisomerase II interaction and apoptotic activity,
although correlation between these events and cytotoxic consequences has not been proved.
Conclusions: The biological response of the cells to the structurally similar anthracyclines may be variable and probably
depends on the cell type which seems to be an additional problem in the multifactorial resistance of tumor cells to anthra-
cyclines.
Key words: apoptosis, anthracyclines, topoisomerase II poisons, cytotoxicity
Comparison of two methods used for monitoring low-copy
cytomegalovirus infection in a patient
with chronic myeloid leukemia
after unrelated umbilical cord blood transplantation
Tomasz Dzieciątkowski1, Maciej Przybylski1, Agnieszka Tomaszewska2, 3,
Małgorzata Rokicka2 and Mirosław Łuczak1
1 Chair and Department of Medical Microbiology, Medical University of Warsaw, Warsaw, Poland
2 Department of Hematology, Oncology and Internal Medicine, Medical University of Warsaw, Warsaw, Poland
3 Department of Hemopoietic Stem Cell Transplantation, Institute of Hematology and Transfusion Medicine, Warsaw, Poland
Received: 2006.08.31, Accepted: 2007.02.28, Published online first: 2007.06.08
Abstract
Introduction: Detection of human cytomegalovirus (CMV, HHV-5) DNA in clinical specimens is considered a cornerstone
in the diagnosis of HHV-5 disease. The present study compared two quantitative methods used for diagnosing cyto-
megalovirus infection in a 21-year-old woman with chronic myeloid leukemia after an unrelated umbilical cord blood trans-
plantation.
Materials and Methods: Blood samples were tested for the presence of HHV-5 DNA using the LightCycler PCR, the quanti-
tative Eclipse® CMV DNA Detection Kit, and a qualitative in-house PCR assay using primers that amplify part of the HHV-5
MIE gene.
Results: Results from samples containing a low cytomegalovirus load were more accurate with the LightCycler test than those
obtained with the Eclipse® test, which underestimated the viral load of samples containing low DNA copy numbers.
Conclusions: These findings underline the value of novel PCR methods used in current therapeutic procedures and in mon-
itoring antiviral therapy with nucleoside analogs. The high level of sensitivity, specificity, accuracy, and rapidity provided by
the LightCycler instrument are favorable for the use of this system in the detection of HHV-5 DNA in clinical specimens.
Key words: chronic myeloid leukemia, umbilical cord blood transplantation, cytomegalovirus infection, real-time PCR.
Serological classification and epitope specificity
of Proteus vulgaris TG 251 from
Proteus serogroup O65
Krystyna Zych1, Katarzyna Kołodziejska1, Dominika Drzewiecka1, Andrey V. Perepelov2,
Yuriy A. Knirel2 and Zygmunt Sidorczyk1
1 Department of General Microbiology, Institute of Microbiology and Immunology, University of £ódŸ, Poland
2 N.D. Zelinsky Institute of Organic Chemistry, Russian Academy of Sciences, Moscow, Russia
Received: 2006.06.13, Accepted: 2006.09.18, Published online first: 2007.06.08
Abstract
Introduction: Proteus rods are currently subdivided into five named species, i.e. Proteus mirabilis, P. vulgaris, P. penneri,
P. hauseri, and P. myxofaciens, and three unnamed Proteus genomospecies 4 to 6. Based on the serospecificity of the
lipopolysaccharide (LPS; O-antigen), strains of P. mirabilis and P. vulgaris were divided into 49 O-serogroups and 11 addi-
tional O-serogroups were proposed later. About 15 further O-serogroups have been proposed for the third medically impor-
tant species, P. penneri. Here the serological classification of P. vulgaris strain TG 251, which does not belong to these
serogroups, is reported. Serological investigations also allowed characterization of the epitope specificity of its LPS.
Materials and Methods: Purified LPSs from five Proteus strains were used as antigens in enzyme immunosorbent assay (EIA),
SDS/PAGE, and Western blot and alkali-treated LPSs in the passive immunohemolysis (PIH) test, inhibition of PIH and
EIA, and absorption of the rabbit polyclonal O-antisera with the respective LPS.
Results: The serological studies of P. vulgaris TG 251 LPS indicated the identity of its O-polysaccharide with that of P. pen-
neri O65. The antibody specificities of P. vulgaris TG 251 and P. penneri O65 O-antisera, were described.
Conclusions: P. vulgaris TG 251 was classified to the Proteus O65 serogroup. Two disaccharide-associated epitopes present in
P. vulgaris TG 251 and P. penneri O65 LPSs are suggested to be responsible for cross-reactions with three heterologous
Proteus strains.
Key words: Proteus, lipopolysaccharide, O-serogroup, epitope, serological classification.
Skin-induced tolerance and its reversal
by Toll-like receptor ligands
Marian Szczepanik
Department of Human Developmental Biology, Jagiellonian University, College of Medicine, Kraków, Poland
Received: 2006.11.22, Accepted: 2006.04.02, Published online first: 2007.06.08
Abstract
In 1970, Gershon was the first to propose that T cells, in addition to their helper activity, can also play a role as regulatory
cells capable of suppressing immune responses. However, the initial strong interest in T cell-mediated suppression was fol-
lowed by a period of doubt and skepticism. Since the late 1990s the “S” word started to be used in immunology again and
interest in T suppressor cells has grown, ushering in a new renaissance for the field. In this article the author presents the
current knowledge about a new subject called “skin-induced tolerance”. Suppression is induced via epicutaneous immuniza-
tion and is described in both Th1- and Tc1-mediated contact sensitivity reactions. The subject of skin-induced tolerance is
also considered in the regulation of experimental models of autoimmune diseases such as allergic autoimmune
encephalomyelitis and collagen-induced arthritis and finally in an animal model of graft rejection. The last part of this pre-
sentation will introduce the very fresh subject of “contrasuppression”, or the reversal of skin-induced suppression.
Key words: suppression, contrasuppression, Toll-like receptors, contact sensitivity, autoimmunity.
M cells are involved in pathogenesis of human contact
lens-associated giant papillary conjunctivitis
Xingwu Zhong1, Hongshan Liu1, Aijun Pu1, Xuefeng Xia2 and Xiaodong Zhou2
1 Zhongshan Ophthalmic Center and State Key Laboratory of Ophthalmology, Sun Yat-sen University,
Guangzhou 510060, China
2 Department of Medicine, University of Texas Health Science Center at Houston, TX, USA
Received: 2006.11.29, Accepted: 2006.02.21, Published online first: 2007.06.08
Abstract
Introduction: The objective was to study the pathogenesis of contact lens-associated giant papillary conjunctivitis (CL-GPC).
Materials and Methods: Twenty-one biopsies of conjunctival giant papillae were obtained from soft contact lens wearers.
The tissues were fixed in 4% paraformaldehyde and embedded in paraffin. Sections of 5 µm thickness were used for studies
of histology and immunohistochemistry of pan-B and pan-T cell distributions.
Results: Conjunctival epitheliums on the top of conjunctiva-associated lymphoid tissue typically lacked goblet cells.
Lymphocytes from underlying lymphoid follicle were pressed into intra-epithelial “pockets” formed through epithelial
invagination. Under the follicle-associated epithelium, pan-B cells were mostly gathered in the central folliclar area and
intraepithelial pockets, while CD3-positive T cells were predominantly distributed in parafolliclar region, but only a few in
the intraepithelial pockets.
Conclusions: Membranous epithelial cells (M cells) play a key role in the pathogenesis of CL-GPC for the binding and
translocation of antigen and pathogen.
Key words: M cells, contact lens-associated giant papillary conjunctivitis
The innate cellular responses to HIV-1 invasion:
emerging molecules of ancient defense mechanisms
Malgorzata Simm
Molecular Virology Division, St. Luke’s/Roosevelt Hospital Center, Columbia University, New York, NY 10019, USA
Received: 2006.12.14, Accepted: 2007.02.22, Published online first: 2007.06.08
Abstract
Along alternative protective pathways, human cells can synthesize biologically active proteins that interfere with HIV replica-
tion, but are not viral antigen specific. HIV is sensitive to several viral inhibitors of cellular origin, such as interferons or inter-
feron-regulated proteins. With the progress of AIDS research it has become evident that the immune cells of some individu-
als are capable of restricting the virus by secretion of other, yet unidentified factor(s) that can be detected only by their potent
antiviral activity. Research efforts to identify this inhibitor of HIV – a “magic bullet” released by our immune cells – have
revealed the identity of several novel molecules and added to the knowledge of innate cellular responses to viral invasion.
Key words: innate immunity, antiviral factors, CD4+ T cells, CD8+ T cells.
Trafficking of FoxP3+ regulatory T cells: myths and facts
Chang H. Kim
Department of Comparative Pathobiology, Purdue Cancer Center, Purdue University, West Lafayette, IN 47907, USA
Received: 2007.01.07, Accepted: 2007.02.22, Published online first: 2007.06.08
Abstract
Fork head box P3 (FoxP3+) regulatory T cells (Tregs) are specialized T cells for prevention of hyperimmune responses and
autoimmunity. Tumors and pathogens can hijack FoxP3+ Tregs to evade host immune responses. There is an increasing body
of evidence that trafficking of FoxP3+ Tregs is important for their effective suppression of target cells. Because of their dis-
tinctive functions and gene expression phenotype, the migratory behavior of FoxP3+ Tregs has been somewhat mystified.
The myths are that they have unique trafficking receptors and migratory behaviors that are different from those of conven-
tional T cells. Another related myth is that FoxP3+ regulatory T cell subsets have a fixed trafficking behavior from the time
they are generated in the thymus. The recent progress in trafficking receptors and migratory behavior of FoxP3+ Tregs is
reviewed here and the validity of these myths is examined.
Key words: FoxP3, regulatory T cells, migration, chemokine receptor, integrin, selectin.
Selected technologies to control genes and their
products for experimental and clinical purposes
Helen K. Alexander1*, Evan P. Booy1, 3*, Wenyan Xiao1, Peyman Ezzati1,
Heinrich Baust4 and Marek Los1, 2, 3
1 Manitoba Institute of Cell Biology, Cancer Care Manitoba, Winnipeg, Canada
2 Department of Human Anatomy and Cell Science, University of Manitoba, Winnipeg, Canada
3 Department of Biochemistry and Medical Genetics, University of Manitoba, Winnipeg, Canada
4 Department of Radiooncology, University of Erlangen, Germany
Received: 2006.12.27, Accepted: 2007.03.15, Published online first: 2007.06.08
Abstract
“On-demand” regulation of gene expression is a powerful tool to elucidate the functions of proteins and biologically-active
RNAs. We describe here three different approaches to the regulation of expression or activity of genes or proteins.
Promoter-based regulation of gene expression was among the most rapidly developing techniques in the 1980s and 1990s.
Here we provide basic information and also some characteristics of the metallothionein-promoter-based system, the tet-off
system, Muristerone-A-regulated expression through the ecdysone response element, RheoSwitch®, coumermycin/novo-
biocin-regulated gene expression, chemical dimerizer-based promoter activation systems, the “Dual Drug Control” system,
“constitutive androstane receptor”-based regulation of gene expression, and RU486/mifepristone-driven regulation of pro-
moter activity. A large part of the review concentrates on the principles and usage of various RNA interference techniques
(RNAi: siRNA, shRNA, and miRNA-based methods). Finally, the last part of the review deals with historically the oldest,
but still widely used, methods of temperature-dependent regulation of enzymatic activity or protein stability (temperature-
-sensitive mutants). Due to space limitations we do not describe in detail but just mention the tet-regulated systems and also
fusion-protein-based regulation of protein activity, such as estrogen-receptor fusion proteins. The information provided
below is aimed to assist researchers in choosing the most appropriate method for the planned development of experimental
systems with regulated expression or activity of studied proteins.
Key words: Dicer, Drosha, FKBP, gene silencing, Mifepristone, mutagensis, N-degron, NF-κB, RISC
Cancer-associated retinopathy
in patients with breast carcinoma
Marta Misiuk−Hojło1, Maria Ejma2, Wojciech A. Gorczyca3, Stanisław Szymaniec3,
Danuta Witkowska3, Wojciech Fortuna3, Ryszard Międzybrodzki3,
Jadwiga Rogozińska−Szczepka4 and Wiesława Bartnik4
1 Department of Ophthalmology, Wroc³aw Medical University, Poland
2 Department of Neurology, Wroc³aw Medical University, Poland
3 Institute of Immunology and Experimental Therapy, Polish Academy of Sciences, Wroc³aw, Poland
4 Institute of Oncology, Gliwice, Poland
Received: 2007.02.02 Accepted: 2007.05.28, Published online first: 2007.07.23
Abstract
Introduction: Cancer-associated retinopathy (CAR) is a paraneoplastic neurological syndrome resulting in progressive loss of
vision and clinical signs of retinal degeneration. It is associated with various types of cancer and is also considered to be an
autoimmune disorder that involves cross-reaction between autoantibodies and retinal proteins. The aim of this study was to
establish whether immunoreactivity to retinal antigens (RAs) observed in patients with breast cancer is accompanied by any
visual impairments.
Materials and Methods: Sera of 295 patients with diagnosed breast cancer were screened for the presence of anti-RAs anti-
bodies using immunoblotting. Cellular immunoreactivity to RAs present in retinal extracts and to purified recoverin and
arrestin was determined by means of a lymphocyte proliferation assay. Six patients with high-titer antibodies to RAs then
underwent ophthalmic and neurological examinations.
Results: Four serum samples contained high-titer antibodies to a 46-kDa protein, most probably retinal α-enolase, three had
antibodies to a 48-kDa protein identified as retinal arrestin, while 56-, 43-, 41-, and 34-kDa antigens were recognized only
by one serum sample each. Moreover, weak cellular response to all the RAs tested was observed in one patient and anoth-
er patient responded only to retinal extract. Two of the examined patients displayed symptoms of CAR.
Conclusions: Immunoreactivity to RAs in patients with breast cancer may also be present in cases without clinical signs of CAR.
Key words: retinal antigens, autoantibodies, breast cancer, visual disorders.
Endothelial progenitor cells as a new agent
contributing to vascular repair
Ewa Miller−Kasprzak and Paweł P. Jagodziński
Department of Biochemistry and Molecular Biology, Karol Marcinkowski University of Medical Sciences, Poznañ, Poland
Received: 2006.10.18, Accepted: 2007.04.26, Published online first: 2007.07.23
Abstract
A special type of stem cells, defined as endothelial progenitor cells (EPCs), has been found in the bone marrow and periph-
eral blood. These EPCs are incorporated into injured vessels and become mature endothelial cells during re-endothelializa-
tion and neovascularization processes. Though a complete phenotypic description of EPCs remains unclear, these cells
express several surface markers, the most relevant including CD34 and CD133 antigens. Furthermore, EPCs derived from
other sources could also give rise to mature endothelial cells, which makes this group of cells more diverse. The recruitment
of EPCs from the bone marrow to homing sites of vasculogenesis is subject to regulation by many factors, including
chemokines and growth factors. The precise mechanism of EPC mobilization and differentiation is not entirely elucidated
and is still under investigation. Recent studies have suggested that EPCs may promote local angiogenesis by secreting angio-
genic growth factors in a paracrine manner. The number and function of EPCs can be affected during pathological condi-
tions, including diabetes mellitus, cardiovascular risk factors for ischemic disease, and graft vasculopathy. Additionally, EPC
number and migration capacity could be improved by such factors as drugs, physical exercise, and growth factors.
Transplantation of EPCs into ischemic tissues may emerge as a promising approach in the therapy of diseases associated with
blood vessel disorders.
Key words: endothelial progenitor cells, angiogenesis, colony-forming unit, surface markers.
Negative signals from FcεεRI engagement
attenuate mast cell functions
Rosa Molfetta, Giovanna Peruzzi, Angela Santoni and Rossella Paolini
Department of Experimental Medicine, University “La Sapienza”, 00161 Rome, Italy
Received: 2007.03.01, Accepted: 2007.05.22, Published online first: 2007.07.23
Abstract
Mast cells have long been recognized as the critical tissue-based effector cells in IgE-mediated allergic diseases. Ligation of
the high-affinity receptor for IgE (FcεRI), constitutively expressed on mast cells, promotes cell activation and immediate
release and production of pro-inflammatory mediators. Besides these positive signals, FcεRI aggregation has recently been
understood to generate negative intracellular signals capable of limiting mast cell functional responses. This review is aimed
at providing a summary of the mechanisms through which FcεRI engagement can generate negative signals and regulate
mast-cell function. Similar mechanisms are employed by other receptors expressed by immune cells, such as T cell and B cell
receptors, pointing to a general concept in negative immunoreceptor signaling.
Key words: mast cell, FcεRI, signal transduction, negative regulation.
Tumor resistance to CD8+ T cell-based
therapeutic vaccination
Yujun Huang*, Shivanee Shah and Liang Qiao
Department of Microbiology and Immunology, Stritch School of Medicine, Loyola University Chicago, Maywood, IL
60153, USA
Received: 2007.01.18, Accepted: 2007.04.28, Published online first: 2007.07.23
Abstract
CD8+ cytotoxic T lymphocytes (CTLs) play an important role in antitumor immunity. Induction of tumor-specific CTLs is
one major strategy for tumor immunotherapy. However, therapeutic vaccinations used to treat firmly established tumors are
generally ineffective. A thorough understanding of the mechanisms underlying tumor resistance to CTL-based therapeutic
vaccination is very important in the tumor immunology field. There are two main mechanisms by which tumors develop resis-
tance to CTL-based therapeutic vaccinations. One is that tumors induce peripheral tolerance of tumor-specific CD8+ T cells.
The other is that tumor cells themselves develop immune evasion mechanisms to prevent recognition and killing by CTLs.
This review focuses on recently reported cellular and molecular mechanisms of CD8+ T cell tolerance and immune evasion
in tumors and discusses about the possibilities to improve tumor immunotherapy.
Key words: CTL, therapeutic vaccination, tumor immunotherapy.
Regulation of T cell homeostasis by JAKs and STATs
Jeremy A. Ross1, Zsuzsanna S. Nagy1, Hanyin Cheng2, Stanislaw M. Stepkowski3
and Robert A. Kirken1
1 University of Texas at El Paso, Department of Biological Sciences, El Paso, TX, USA
2 University of Texas Health Science Center at Houston, Department of Integrative Biology and Pharmacology, Houston,
TX, USA
3 University of Texas Health Science Center at Houston, Department of Surgery, Division of Immunology
and Organ Transplantation, Houston, TX, USA
Received: 2007.06.18, Accepted: 2007.07.06, Published online first: 2007.07.23
Abstract
Regulation of T cell homeostasis is critical for maintaining normal immune function. An imbalance in T cell proliferation
can result in disorders ranging from cancer and autoimmunity to immunodeficiencies. Full activation of T cells requires three
sequential signals, where signal 3, which is delivered by multiple cytokines, regulates proliferation, differentiation, and sur-
vival/death. Signaling from cytokines through their receptors is primarily delivered by two molecular families, namely Janus
tyrosine kinases (JAKs) and signal transducers and activators of transcription (STATs). Invaluable knowledge about JAKs
and STATs has arisen from studies of mice made genetically deficient in these molecules, analyses of tumor models, and
studies of expression patterns by proteomics/genomics, which all have begun to define the role of JAKs and STATs in sur-
vival versus apoptosis. These findings also have suggested ways in which JAKs and STATs may be manipulated for thera-
peutic intervention in lymphoid-derived diseases. This review seeks to focus on the role of JAK tyrosine kinases and STAT
transcription factors in mediating the lymphocyte life cycle and how they might be manipulated for therapeutic applications.
Key words: T cells, cytokines, Janus tyrosine kinases (JAK), signal transducers and activators of transcription (STAT), apop-
tosis, disease.
Anti-tumor action of tumor necrosis factor
against Bomirski Ab melanoma in hamsters
Patrycja Koszałka1, Ewa Szmit2, Andrzej Myśliwski2 and Jacek Bigda1
1 Department of Medical Biotechnology, Division of Cell Biology, Intercollegiate Faculty of Biotechnology,
Medical University of Gdañsk, Gdañsk, Poland
2 Department of Histology and Immunology, Medical University of Gdañsk, Gdañsk, Poland
Received: 2007.01.04, Accepted: 2007.05.27, Published online first: 2007.07.23
Abstract
Introduction: Tumor necrosis factor (TNF) is a cytokine able to exert anti-tumor activity in various models and modes of
applications. However, the exact mechanism mediating the in vivo anti-tumor effect of TNF has not yet been clarified.
Materials and Methods: The effects of intratumoral injection of rat TNF into hamsters bearing Bomirski Ab amelanotic
melanoma, a fast growing tumor of high metastatic potential, were tested. Subcutaneous injections of the anti-angiogenic
compound TNP-470 allowed analysis of its influence on the effects of TNF administration.
Results: TNF application resulted in a significant inhibition of tumor growth and changes in metastasis pattern. Accelerated
hemorrhagic necrosis was also observed, indicating the effect of the cytokine on tumor vessels. Moreover, the synergistic
anti-tumor effect of TNF and anti-angiogenic agent TNP-470 suggested a cooperative activity of both substances on tumor
vasculature. Microscopically, the effect of TNF injections was expressed by an increase in the amount of tumor cells with
nuclear pyknosis and karryorrhexis. In vitro assays indicated a direct cytotoxic effect of TNF against Ab melanoma cells, most
probably as an outcome of apoptosis. Intratumoral application of TNF also caused some modulation of cytokine response in
melanoma-bearing hamsters as evidenced by increased levels of IL-6 in blood serum.
Conclusions: This study established Bomirski Ab melanoma as a useful model for complex analysis of the anti-tumor activity
of TNF.
Key words: TNF, melanoma, tumor vasculature, immune modulation.
The function of interleukin 17
in the pathogenesis of rheumatoid arthritis
Agnieszka Paradowska1, Włodzimierz Maśliński2, Agnieszka Grzybowska−Kowalczyk1
and Jan Łącki1, 3
1 Department of Biochemistry, Institute of Rheumatology, Warsaw, Poland
2 Department of Pathophysiology and Immunology, Institute of Rheumatology, Warsaw, Poland
3 Department of Rheumatic Diseases, Institute of Rheumatology, Warsaw, Poland
Received: 2007.03.14, Accepted: 2007.05.28, Published online first: 2007.10.01
Abstract
Interleukin (IL)-17 is a 30- to 35-kDa homodimeric polypeptide cytokine cloned in 1993 and originally named cytotoxic T
lymphocyte-associated antigen-8 (CTLA-8). Sequencing the human genome resulted in the discovery of an additional five
members of the IL-17 family that were consecutively named IL-17B to IL-17F. IL-17A is exclusively produced by a newly
identified CD4+ T-helper subset that was recently named Th17. Differentiation of these cells from naïve CD4+ T cells requires
both TGF−β and IL-6. IL-15 and, especially, IL-23 are required for these cells’ survival and efficient IL-17 production. IL-17
binding to an IL-17 receptor expressed on epithelial, endothelial, and fibroblastic stromal cells triggers the activation of tran-
scription factor NF-κB and mitogen-activated protein kinase (p-38), which in turn results in the secretion of IL-1, TNF-α,
IL-6, IL-8, or prostaglandin E2. The IL-17 family plays a key role in the regulation of immune and inflammatory response,
in the homeostasis of several tissues, and the progression of autoimmune diseases. In addition, IL-17 exerts synergistic effects
with TNF-α and IL-1 in the induction of joint inflammation and cartilage and joint destruction. Given these properties, it is
not surprising that in certain pathological conditions, for example rheumatoid arthritis, Th17 cells emerge as a new patho-
logical cell type that, by IL-17 production and release, contributes to their pathogeneses.
Key words: interleukin 17, family and receptors of IL-17, function of IL-17, rheumatoid arthritis.
Evaluation of selected parameters
of the antioxidative system in patients with type 2
diabetes in different periods of metabolic compensation
Jacek Rysz1, Robert Błaszczak1, Maciej Banach2, Kornelia Kędziora−Kornatowska3,
Tomasz Kornatowski4, Wojciech Tański5 and Józef Kędziora6
1 2nd Department of Family Medicine, University Hospital No. 2, Medical University of £ódŸ, Poland
2 Department of Cardiology, 1st Chair of Cardiology and Cardiac Surgery, University Hospital No. 3,
Medical University of £ódŸ, Poland
3 Department and Clinic of Geriatrics, Medical Academy of Bydgoszcz, Poland
4 Department of Pharmacology and Therapy, Medical Academy of Bydgoszcz, Poland
5 Department of Internal Medicine, Military Hospital of Wroc³aw, Poland
6 Department of Clinical Chemistry and Biochemistry, Medical University of £ódŸ, Poland
Received: 2006.10.19, Accepted: 2007.04.02, Published online first: 2007.10.01
Abstract
Introduction: The aim of the study was to evaluate some selected parameters of the antioxidative system in patients with type
2 diabetes.
Materials and Methods: Eighty-one type 2 diabetes patients took part in the study (39 patients with metabolically balanced and
42 with metabolically unbalanced diabetes). The control group consisted of 30 healthy people. The total antioxidant capacity of
plasma was measured fluorometrically using phycoerythrin. To calculate the low-molecular-weight antioxidant concentration in
the plasma samples, the duration of Trolox activity as a function of its concentration in the sample was measured. The activity
of antioxidative enzymes in red blood cells was determined using the Misra and Fridovich method and Beers and Sizer method.
Results: The total plasma antioxidant capacity and the low-molecular-weight antioxidant concentration in the group of
patients with metabolically compensated type 2 diabetes were statistically significantly higher than in the group of patients
with metabolically uncontrolled diabetes. The activity of antioxidative enzymes was found to be higher in the group of type
2 diabetes patients at the stage of metabolic balance.
Conclusions: The obtained results confirm the thesis of glucose toxicity and intensification of oxidative stress in patients with
diabetes.
Key words: oxidative stress, diabetes mellitus, antioxidative enzyme.
Dendritic/tumor fusion cell-based vaccination
against cancer
Shigeo Koido1, 2, Eiichi Hara3, Sadamu Homma4, Kiyotaka Fujise1, 2, Jianlin Gong5
and Hisao Tajiri1
1 Division of Gastroenterology and Hepatology, Department of Internal Medicine,
The Jikei University School of Medicine, Tokyo, Japan
2 Institute of Clinical Medicine and Research, The Jikei University School of Medicine, Tokyo, Japan
3 Saitama Cancer Center Research Institute for Clinical Oncology, Saitama, Japan
4 Department of Oncology, Institute of DNA Medicine, The Jikei University School of Medicine, Tokyo, Japan
5 Department of Medicine, Boston University School of Medicine, Boston, MA, USA
Received: 2007.03.26, Accepted: 2007.05.28, Published online first: 2007.10.01
Abstract
A promising area of investigation is the use of cancer vaccines to eliminate residual tumor cells. Dendritic cells (DCs) are
potent professional antigen-presenting cells able to induce primary immune responses. DCs capture and process antigens
into peptides and present them to T cells and B cells through MHC class I and II molecules. An alternative approach to the
induction of antitumor immunity is the use of fusions of DCs and tumor cells. In this approach, a broad spectrum of tumor-
-associated antigens, including those known and unidentified, are processed endogenously and presented by MHC class I and
II pathways in the context of costimulatory signals. In animal studies, vaccination with DC/tumor fusion cells results in the
elimination of established lung metastasis. Preclinical human studies have demonstrated that DC/tumor fusion cells induce
antigen-specific polyclonal cytotoxic T-lymphocyte responses against autologous tumor in vitro. In clinical studies, vaccina-
tion of cancer patients with autologous DC/tumor fusion cells is associated with immunological and clinical responses in
a subset of patients. Future studies should be investigated to improve the immunogenicity of DC/tumor fusion cell prepara-
tions. This review provides a general overview of the DC/tumor fusion cell-based vaccine and summarizes some of the recent
advances in this field.
Key words: dendritic cells, fusion cell vaccine, tumor immunity, antigen-specific CTL.
Serum concentration of C-reactive protein
is not a good marker of bronchial hyperresponsiveness
Bernard Panaszek, Ewa Liebhart, Jerzy Liebhart, Robert Pawłowicz and Andrzej M. Fal
Department of Internal Medicine and Allergology, Wroc³aw Medical University, Wroc³aw, Poland
Received: 2007.01.04, Accepted: 2007.05.28, Published online first: 2007.10.01
Abstract
Introduction: Asthmatic inflammation is responsible for vital features of the disease, including bronchial hyperresponsiveness
(BHR). At present we do not have precise markers for monitoring asthmatic inflammation. C-reactive protein (CRP),
a marker of systemic inflammation, seemed to be a factor which could also reflect the level of asthmatic inflammation
expressed by BHR. Therefore the relationship between CRP concentration and BHR was evaluated.
Materials and Methods: One hundred and two patients entered the study. A skin prick test with a broad spectrum of common
aeroallergens as well as baseline spirometry and a histamine bronchoprovocation test were performed in each subject. Blood
samples for high-sensitivity CRP (hsCRP) measurement were taken before the bronchial challenge tests.
Results: Serum hsCRP concentrations ranged from 0.20 to 14.5 mg/l (median: 1.2 mg/l, 25–75% quartiles: 0.6–2.4). Positive
skin prick tests were found in 26 subjects. Bronchial hyperresponsiveness was confirmed in 42 patients (first subgroup), while
60 subjects did not demonstrate BHR (second subgroup). Among the patients with BHR, asthma was diagnosed in 33 cases
and Corrao syndrome in 9. In both subgroups, serum hsCRP concentrations had similar levels (median: 1.4 mg/l, 25–75%
quartiles: 0.8–2.4 and median: 0.9 mg/l, 25–75% quartiles: 0.5–2.8, respectively; p=0.297). There was no statistically signifi-
cant correlation (r= –0.163, p=0.302) between serum hsCRP concentration and the level of BHR expressed as the 20%
provocative concentration for histamine. In addition, hsCRP serum concentration, after adjustment for age, atopy, body
mass index, and gender, was not a significant predictor of positive histamine bronchoprovocation test results (p=0.22,
OR=0.86, 95% CI).
Conclusions: Serum hsCRP concentration is not a good marker of BHR, which is mainly dependent on asthmatic inflamma-
tion and is measured during bronchial challenge with histamine. This finding is important for interpreting and discussing
results obtained from epidemiological and population-based studies on relationships between either CRP concentration and
BHR or local and systemic inflammation.
Key words: asthmatic inflammation, asthma, Corrao syndrome, BHR, high-sensitivity CRP, systemic inflammation.
Mechanisms by which selenium
influences immune responses
Peter R. Hoffmann
Department of Cell and Molecular Biology, John A. Burns School of Medicine, University of Hawai’i, Honolulu,
HI 96813, USA
Received: 2007.06.28, Accepted: 2007.09.10, Published online first: 2007.10.01
Abstract
Selenium (Se) is an essential dietary trace element that influences immune responses through its incorporation into seleno-
proteins as the amino acid selenocysteine. This review summarizes data available to date regarding the mechanisms by which
Se exerts its effects on inflammation and immune responses. This includes the effects of Se on phagocytes as well as effects
on lymphocyte activation, proliferation, and differentiation. Also examined are the known functions of individual seleno-
proteins for regulating reactive oxygen species and redox potential in leukocytes. Overall, determining how Se contributes to
optimal immune responses will depend on a better understanding of the mechanisms by which the selenoproteins, individu-
ally and collectively, shape inflammation and immune responses.
Key words: selenium, selenoproteins, immune responses, inflammation, oxidative stress.
The immunological burden
of human cytomegalovirus infection
Naeem Khan
Division of Immunology, School of Infection and Host Defence, University of Liverpool, UK
Received: 2007.07.31, Accepted: 2007.09.10, Published online first: 2007.10.01
Abstract
Cytomegalovirus (CMV) is a persistent DNA virus that has evolved with humans to establish a finely balanced host-virus
relationship. This balance is maintained by host immune surveillance since deficiencies in these processes can result in life-
-threatening disease, as observed in immunologically immature neonates and pharmacologically immunosuppressed trans-
plant recipients. Both T cells and natural killer cells are intimately involved in maintaining asymptomatic infection by spe-
cific and non-specific recognition of infected cells. Under pressure from such host immune responses, CMV appears to have
evolved elaborate strategies to subvert these responses in order to persist in the host. CMV target antigens are well charac-
terized, with many CD8 T cell and CD4 T cell epitopes reported. This information is now being exploited to treat immuno-
compromised patients in order to boost virus-specific immunity. This review also discusses our current understanding of how
virus carriage may skew lymphocyte populations in immunocompetent subjects and the association of CMV-seropositivity
with immunosenescence.
Key words: cytomegalovirus, immune responses, therapy, immunosenescence
Approaches and challenges
in targeting memory T cells in transplant tolerance
Ming Chen and Xian Chang Li
Transplant Research Center, Beth Israel Deaconess Medical Center and Harvard Medical School, Boston, MA 02215, USA
Received: 2007.08.03, Accepted: 2007.09.10, Published online first: 2007.10.01
Abstract
Memory T cells are an important cell type in the immune system and are vital to protective immunity against invading
pathogens. However, a significant fraction of memory T cells is found to be alloreactive in transplant models, i.e. they can
readily attack and dismantle allografts in transplant models. As memory T cells are not as easily amenable as naïve T cells,
memory T cells constitute a potent barrier to the induction of transplant tolerance. The key issues concerning memory T cells
in transplantation are related to the tolerability of alloreactive memory T cells and the effects of commonly used immuno-
suppressive drugs on the memory response in transplant recipients. The real challenge in the future is to selectively tolerize
alloreactive memory T cells but spare those involved in protective immunity following organ transplantation. This review will
discuss recent advances in our understating of memory T cells in transplant models, with specific emphasis on the problems
and challenges in targeting memory T cells in the induction of transplant tolerance.
Key words: transplantation, rejection, tolerance, memory, alloreactivity.
Terpenes: substances useful in human healthcare
Roman Paduch1, Martyna Kandefer−Szerszeń1, Mariusz Trytek2 and Jan Fiedurek2
1 Department of Virology and Immunology, Institute of Microbiology and Biotechnology,
Maria Curie-Sk³odowska University, Lublin, Poland
2 Department of Industrial Microbiology, Institute of Microbiology and Biotechnology,
Maria Curie-Sk³odowska University, Lublin, Poland
Received: 2007.01.23, Accepted: 2007.05.10, Published online first: 2007.10.01
Abstract
Terpenes are naturally occurring substances produced by a wide variety of plants and animals. A broad range of the biolog-
ical properties of terpenoids is described, including cancer chemopreventive effects, antimicrobial, antifungal, antiviral, anti-
hyperglycemic, anti-inflammatory, and antiparasitic activities. Terpenes are also presented as skin penetration enhancers and
agents involved in the prevention and therapy of several inflammatory diseases. Moreover, a potential mechanism of their
action against pathogens and their influence on skin permeability are discussed. The major conclusion is that larger-scale use
of terpenoids in modern medicine should be taken into consideration.
Abbreviations: 5-FU – 5-fluorouracil, AKBA – acetyl-11-keto-β-boswellic acid, AZT – zidovudine, CoQ – coenzyme Q, COX-2
– cyclooxygenase 2, DMAPP – dimethylallyl pyrophosphate, HIV – human immunodeficiency virus, HSV – herpes simplex
virus, iNOS – inducible nitric oxide synthetase, IPP – isopentenyl pyrophosphate, LPS – lipopolysaccharide, NF-κB – nuclear
factor κB, NO – nitric oxide, PGE2 – prostaglandin E2, PLA2 – phospholipase A2, TGF-β – transforming growth factor β,
TNF-α – tumor necrosis factor α.
Key words: terpenes, terpene activity, antitumor, antimicrobial, antifungal, antiviral, antihyperglycemic, anti-inflammatory,
antiparasitic.
Serological and structural characterization
of the O-antigens of the unclassified
Proteus mirabilis
strains TG 83, TG 319, and CCUG 10700 (OA)
Agnieszka Zabłotni1, Krystyna Zych1, Anna N. Kondakova2, Małgorzata Siwińska1,
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, Moscow, Russian Federation
Received: 2006.12.11, Accepted: 2007.05.10, Published online first: 2007.10.01
Abstract
Introduction: Lipopolysaccharide (endotoxin, LPS) is an important potential virulence factor of Proteus rods. The serological
specificity of the bacteria is defined by the structure of the O-polysaccharide chain (O-antigen) of the LPS. Until now, 76
O-serogroups have been differentiated among Proteus strains.
Materials and Methods: LPSs were isolated from Proteus mirabilis TG 83, TG 319, and CCUG 10700 (OA) strains by phe-
nol/water extraction. Antisera were raised by immunization of rabbits with heat-killed bacteria. Serological investigations
were performed using enzyme immunosorbent assay, passive immunohemolysis, inhibition of both assays, absorption of anti-
sera, and Western blot.
Results: The cross-reactive epitope shared by these strains and P. penneri O72a,O72b is located on the O-polysaccharide and
is most likely associated with an α-D-Glcp-(1→6)-β-D-GalpNAc disaccharide fragment. The serological data indicated the
occurrence of two core types in the LPSs studied, one characteristic for P. mirabilis TG 319 and CCUG 10700 (OA) and the
other for P. mirabilis TG 83 and O57.
Conclusions: The serological and structural data showed that P. mirabilis TG 83, TG 319, CCUG 10700 (OA), and O57 have
the same O-antigen structure and could be qualified to the Proteus O57 serogroup.
Key words: Proteus, lipopolysaccharide, O-serogroup, serological classification.
Regulation of local immunity by airway epithelial cells
Anja K. Mayer and Alexander H. Dalpke
Department of Hygiene and Medical Microbiology, Hygiene Institute, University of Heidelberg, Germany
Received: 2007.07.11, Accepted: 2007.09.18, Published online first: 2007.12.03
Abstract
Epithelial cells are the first line of defense against invading microbial pathogens. They are important contributors to innate
mucosal immunity and generate various and sophisticated anti-microbial defense mechanisms, including the formation of
a tight barrier and secretion of anti-microbial substances as well as inflammatory mediators. To provide these active defense
mechanisms, epithelial cells functionally express various pattern-recognition receptors. Toll-like receptors have been shown
to recognize conserved microbial patterns mediating inducible activation of innate immunity. Mucosal surfaces, however, are
prone to contact with pathogenic as well as non-pathogenic microbes and, therefore, immune-recognition principles have to
be strictly regulated to avoid uncontrolled permanent activation. This review will focus on mechanisms by which epithelial
cells regulate mucosal immune responses, thus creating an organ-specific microenvironment. This includes local adaptations
in microbial recognition, regulation of local immune homeostasis, and modulation of antigen-presenting cells and adaptive
immune responses. These regulatory mechanisms serve the special needs of controlled microbial recognition in mucosal
compartments.
Key words: epithelial cells, immunity, lung, Toll-like receptors.
Alterations in the expression of signal-transducing
CD3ζζ chain in T cells from patients
with chronic inflammatory/autoimmune diseases
Lidia Ciszak1, Edyta Pawlak1, Agata Kosmaczewska1, Stanisław Potoczek2
and Irena Frydecka1, 2
1 Department of Experimental Therapy, Institute of Immunology and Experimental Therapy, Polish Academy of Sciences,
Wroc³aw, Poland
2 Department of Hematology, Wroc³aw Medical University, Poland
Received: 2007.06.19, Accepted: 2007.09.18, Published online first: 2007.12.03
Abstract
The CD3ζ chain, a component of the T cell receptor (TCR)/CD3 complex, is considered to be a limiting factor in the assem-
bly and transport of the TCR/CD3 complex to the cell surface and is crucial to receptor signaling function. Recent studies
have demonstrated altered expression and function of this signal transduction molecule in T and natural killer cells in
patients with chronic inflammatory/autoimmune diseases. In this review, current knowledge concerning the expression of
CD3ζ chain as well as the mechanisms responsible for abnormal expression of this molecule in systemic lupus erythemato-
sus, rheumatoid arthritis, and childhood idiopathic nephrotic syndrome are summarized.
Key words: CD3ζ chain, TCR/CD3 complex, chronic inflammatory diseases, autoimmune diseases
Recent discoveries in the genetics of melanoma
and their therapeutic implications
Amélie Marquette, Martine Bagot, Armand Bensussan and Nicolas Dumaz
INSERM Unit 841, IMRB, Department of Immunology, Dermatology, and Oncology, Créteil, France
Received: 2007.08.31, Accepted: 2007.10.18, Published online first: 2007.12.03
Abstract
The incidence of cutaneous malignant melanoma, tumors arising from melanocytes, has increased markedly over the past
few years in many countries. Although early melanoma is curable through surgical excision, the prognosis of advanced
melanoma is very poor, this tumor being resistant to current therapies. Thus there is a need for new therapies to improve the
treatment of advanced melanoma. This review provides an overview of recent discoveries in the genetics of melanoma which
could offer new therapeutic opportunities.
Key words: melanoma, receptor tyrosine kinase, MAPK pathway, PI3K pathway, cAMP pathway, small molecule inhibitor.
DNA vaccines: are they still just a powerful tool
for the future?
Jana Běláková, Milada Horynová, Michal Křupka, Evžen Weigl and Milan Raška
Department of Immunology, Faculty of Medicine and Dentistry, Palacky University, Olomouc, Czech Republic
Received: 2007.06.25, Accepted: 2007.10.29, Published online first: 2007.12.03
Abstract
Vaccination is historically one of the most successful strategies for the prevention of infectious diseases. For safety reasons,
modern vaccinology tends toward the usage of inactivated or attenuated microorganisms and uses predominantly subunit
vaccines. The antigens need to be clearly defined, pure, stable, appropriately composed, and properly presented to the
immune system of the host. Differing ratios of various proportions between specific CD4+ and CD8+ T cell responses are
essential for conferring the required protection in the case of individual vaccines. To stimulate both CD4+ and CD8+ T cells,
the antigens must be processed and presented to both antigen-presentation pathways, MHC I and MHC II. Protein antigens
delivered by vaccination are processed as extracellular antigens. However, extracellularly delivered antigen can be directed
towards intracellular presentation pathways in conjugation with molecules involved in antigen cross-presentation, e.g. heat
shock proteins, or by genomic-DNA vaccination. In this overview, current knowledge of the host immune response to DNA
vaccines is summarized in the introduction. The subsequent sections discuss techniques for enhancing DNA vaccine effica-
cy, such as DNA delivery to specific tissues, delivery of DNA to the cell cytoplasm or nucleus, and enhancement of the
immune response using molecular adjuvants. Finally, the prospects of DNA vaccination and ongoing clinical trials with var-
ious DNA vaccines are discussed.
Key words: DNA vaccine, plasmid, delivery systems, CTL, MHC I, MHC II.
Immunomodulation by αα(1)-proteinase inhibitor:
lack of chemotactic effects of recombinant human
αα(1)-proteinase inhibitor from yeast
on human peripheral blood granulocytes
Birgit Mosheimer, Reinhard Alzner and Christian J. Wiedermann
Division of General Internal Medicine, Department of Medicine, Medical University of Innsbruck, A-6020 Innsbruck, Austria
Received: 2007.07.24, Accepted: 2007.10.02, Published online first: 2007.12.03
Abstract
Introduction: Recombinant α(1)-proteinase inhibitor, clinically developed for inhalative augmentation therapy in patients
with α(1)-proteinase inhibitor deficiency or cystic fibrosis, may directly contribute to leukocyte accumulation as it may func-
tion as a chemoattractant. The migratory effects of yeast-derived human recombinant α(1)-proteinase inhibitor on human
peripheral blood neutrophils and eosinophils were therefore tested in vitro.
Materials and Methods: Human peripheral blood leukocytes were prepared from forearm venous blood and tested for migra-
tion toward various preparations of yeast-derived recombinant α(1)-proteinase inhibitor in modified Boyden-chamber
micropore filter assays.
Results: No direct effects of yeast-derived recombinant human α(1)-proteinase inhibitor on in vitro migration of isolated neu-
trophils or eosinophils were seen.
Conclusions: The lack of direct chemotactic effects of recombinant human α(1)-proteinase inhibitor despite anti-inflamma-
tory effects in other biological activities of leukocytes may contribute to the preserved antibacterial defense mechanisms
observed in patients under experimental augmentation therapy with inhaled α(1)-proteinase inhibitor.
Key words: neutrophils, eosinophils, chemotaxis, antiproteinase, emphysema.
The influence of different culture microenvironments
on the generation of dendritic cells
from non-small-cell lung cancer patients
Paweł Krawczyk1, Kamila Wojas2, Janusz Milanowski1 and Jacek Roliński2
1 Department of Pneumology, Oncology and Allergology, Medical University of Lublin, Poland
2 Clinical Immunology Department, Medical University of Lublin, Poland
Received: 2007.06.05, Accepted: 2007.10.25, Published online first: 2007.12.03
Abstract
Introduction: Monocyte-derived dendritic cells (DCs) are currently under extensive evaluation as cell vaccines for cancer
treatment. Many protocols regarding DCs generation in vitro with different protein components, especially autologous pro-
teins, have been described. On the other hand, active tumor-derived factors in patients’ serum could impair monocytes,
which might result in their abrogated differentiation into DCs in vitro.
Materials and Methods: Autologous DCs from non-small-cell lung cancer (NSCLC)-bearing patients were generated in dif-
ferent culture microenvironments. Peripheral blood mononuclear cells (PBMCs) were cultured in the presence of inter-
leukin-4 and granulocyte-monocyte-stimulating factor with supplementation of 10% autologous serum, 10% allogenic
serum, or 2% human albumin. The course of apoptosis, phagocytic ability, and the immunophenotype of the generated DCs
were analyzed using flow cytometric methods.
Results: After 48 h of culture, we found a lower percentage of CD1a+/CD14+ and a higher percentage of CD1a+/CD14– cells
in the culture supplemented with human albumin than in the cultures supplemented with serums. The lowest CD14 antigen
expression was found in the human albumin-supplemented 48-h cultures. After 48 h in the cultures carried out with human
albumin we found significantly higher percentages of AV+/PI+ cells and AV–/PI+ cells than in cultures supplemented with
autologous or allogenic serum. We also noted that the expression of FITC-dextran after 4 and 24 h of incubation was sig-
nificantly higher in the cultures supplemented with both serums than in the HA-SC. The percentage of semi-mature DCs
and of CD83 expression was lowest in the culture supplemented with 2% human albumin.
Conclusions: The kind of culture supplementation had a great impact on the apoptosis of cultured PBMCs. It could also influ-
ence the yield of monocyte-derived DCs. It was also confirmed that autologous and allogenic serums provide suitable
microenvironments for the generation of autologous DCs from NSCLC patients. The choice of culture supplementation for
DC generation is still unsolved and further studies should be undertaken.
Key words: dendritic cells, non-small-cell lung cancer, immunophenotyping, cell culture.
Total antioxidant status and 8-hydroxy-2’-deoxyguanosine
levels in gingival and peripheral blood
of periodontitis patients
Tomasz Konopka1, Katarzyna Król2, Wacław Kopeć3 and Hanna Gerber4
1 Department of Oral Pathology, Wroc³aw Medical University, Wroc³aw, Poland
2 Department of General Dentistry, Pomeranian Medical University, Szczecin, Poland
3 Immunological Laboratory of Nephrology Clinic, Wroc³aw Medical University, Wroc³aw, Poland
4 Department of Maxillofacial Surgery, Wroc³aw Medical University, Wroc³aw, Poland
Received: 2007.03.07, Accepted: 2007.10.25, Published online first: 2007.12.03
Abstract
Introduction: The aim of this study was to determine 8-OHdG concentration as a biomarker of oxidant-induced DNA dam-
age and to assess total antioxidant status (TAS) in gingival and peripheral blood during periodontal lesion.
Materials and Methods: The study included 56 untreated periodontitis patients (26 with aggressive periodontitis, and 30 with
chronic periodontitis (CP). The control group consisted of 25 healthy volunteers without pathological changes in the peri-
odontium. Competitive ELISA was used to measure 8-OHdG. A colorimetric method based on the reduction of ABTSo+
radical cation generation was used to measure TAS.
Results: Significantly higher 8-OHdG concentrations were detected in the gingival blood in both groups of patients with peri-
odontitis than in the control group. Subjects with CP had significantly decreased TAS levels in the gingival blood compared
with the control group. A significantly decreased TAS level in the peripheral blood in both patient groups compared with the
control group was found. Significant positive correlation between TAS levels in venous and gingival blood in all the peri-
odontitis patients and in the CP group was observed.
Conclusions: The oxidative burst in periodontitis may lead to significant local damage to nucleic acids. The significantly
decreased TAS level in the gingival blood of CP patients compared with the healthy subjects suggests the possibility of a sig-
nificant decrease in local antioxidant system capacity during the course of periodontitis. The decreased TAS level in the
peripheral blood in the group of all patients with periodontitis may be one of the pathogenic mechanisms underlying the links
between periodontal disease and several systemic diseases for which periodontitis is regarded as a independent risk factor.
Key words: reactive oxygen species, 8-hydroxy-2’-deoxyguanosine, total antioxidant status, oxidative stress, periodontitis.