CONTENTS
Reviews
- Stroke, myocardial infarction, acute and chronic inflammatory diseases: caspases and other apoptotic molecules as targets for drug development
Michael Kreuter1, Claus Langer2, Claus Kerkhoff3, Pallu Reddanna4, Anna L. Kania5, Subbareddy Maddika4,5, Katerina Chlichlia6, Truc Nguyen Bui7 and Marek Los3,5 (1Department of Medicine/Hematology and Oncology, Muenster, Germany 2Central Clinical Laboratory, Medical Clinic, Muenster, Germany 3Institute of Experimental Dermatology, University of Muenster, Germany 4School of Life Sciences, University of Hyderabad, India 5Manitoba Institute of Cell Biology, Winnipeg, Canada 6Department of Tropical Hygiene, University of Heidelberg, Heidelberg, Germany 7Ben May Institute for Cancer Research, University of Chicago, Chicago, IL 60637, USA)Abstract. Mapping of the human and other eukaryotic genomes has provided the pharmacological industry with excellent models for drug discovery. Control of cell proliferation, differentiation, activation and cell removal is crucial for the development and existence of multicellular organisms. Each cell cycle progression, with sequences of DNA replication, mitosis, and cell division, is a tightly controlled and complicated process that, when deregulated, may become dangerous not only to a single cell, but also to the whole organism. Regulation and the proper control of the cell cycle and of programmed cell death (apoptosis) is therefore essential for mammalian development and the homeostasis of the immune system. The molecular networks that regulate these processes are critical targets for drug development, gene therapy, and metabolic engineering. In addition to the primary, intracellular apoptotic suicide machinery, components of the immune system can detect and remove cells and tissue fragments that no longer serve their defined functions. In this review we will focus on apoptotic pathways converging on caspase family proteases, summarizing pharmacological attempts that target genes, proteins, and intermolecular interactions capable of modulating apoptosis and the inflammatory response. The upcoming pharmacological development for treatment of acute pathologies, such as sepsis, SIRS, stroke, traumatic brain injury, myocardial infarction, spinal cord injury, acute liver failure, as well as chronic disorders such as Huntington’s disease, Parkinson’s disease, ALS, and rheumatoid arthritis, will be discussed in details. We also suggest new potential molecular targets that may prove to be effective in controlling apoptosis and the immune response in vivo.
Keywords: apoptosis; Bcl-2, inflammation; myocardial infarct; stroke; sepsis
- Mechanisms of type I interferon signaling in normal and malignant cells
Yongzhong Li, Kishore K. Srivastava and Leonidas C. Platanias Robert H. Lurie (Comprehensive Cancer Center and Division of Hematology-Oncology, Northwestern University Medical School and Lakeside Veterans, Administration Medical Center, Chicago, IL 60611, USA)Abstract. Type I interferons (IFNs) are cytokines that induce multiple biological effects on target cells, including antiviral, antiproliferative, and immunomodulatory activities. Consistent with the pleiotropic nature of these cytokines, multiple signaling pathways are activated during binding of IFNs to the type I IFN receptor. An important signaling cascade activated by type I IFNs is the Jak-Stat pathway. Activation of the Tyk-2 and Jak-1 kinases, and downstream formation of various Stat complexes, mediates IFN-dependent gene transcription for IFN-stimulated genes. In addition to the classic Jak-Stat pathway, type I IFNs activate multiple other pathways, including the insulin receptor substrate-phosphatidylinositol 3’-kinase cascade, the CBL-CrkL pathway, and mitogen-activated protein kinase pathways. There is accumulating evidence that non-Stat IFN-regulated signaling pathways play important roles in the generation of the antiproliferative effects of type I IFNs. In this review, the regulation of various signaling cascades by the type I IFN receptor is summarized and an update on recent advances in the field is provided.
Keywords: Interferon; cytokines; signaling; malignat cells
- Targeting the chemokine network in renal inflammation
Vaclav Eis1,2, Volker Vielhauer1 and Hans-Joachim Anders1 (1Nephrological Center, Medical Policlinic, Ludwig-Maximilians-University Muenich, Germany 2Department of Pathology, 3rd Faculty of Medicine, Charles University, Prague, Czech Republic)Abstract. Chemokines and their receptors are involved in the pathogenesis of renal diseases. They mediate leukocyte recruitment and activation during initiation as well as progression of renal inflammation. Infiltrating leukocyte subpopulations contribute to renal damage by releasing inflammatory and profibrotic cytokines. All intrinsic renal cells are capable of chemokine secretion on stimulation in vitro. Expression of inflammatory chemokines correlates with renal damage and local accumulation of chemokine receptor-bearing leukocytes in a variety of animal models of renal diseases as well as in human biopsy studies. Chemokines and their respective receptors could represent new targets for therapeutic intervention in renal inflammatory disease states that often tend to progress to end-stage renal disease. This article summarizes the present data on the role of chemokines and their receptors in renal inflammation with special emphasis on our efforts to identify the chemokine receptors CCR1 and CCR2 as promising targets for therapeutic intervention.
Keywords: chemokines; inflammation; kidney disease; fibrosis
- Tracking lymphocytes in vivo
Claire L. Adams, Natalie Kobets, Gordon R. Meiklejohn, Owain R. Millington, Angela M. Grierson, Catherine M. Rush, Karen M. Smith and Paul Garside (University of Glasgow, Western Infirmary, Glasgow, Scotland, UK)Abstract. The ability to track antigen (Ag)-specific lymphocyte populations in vivo has greatly increased our understanding of the location and functional status of these cells throughout the course of an immune response. Recent technical advances have enhanced researchers’ capability to follow migration, activation and cellular interactions of Ag-specific lymphocytes in situ. It is now possible to monitor changes in T cell subsets, co-stimulatory molecules, and chemokine expression within the physiological context of secondary lymphoid organs. Furthermore, the Ag-presenting cell-T cell interaction can be studied, thus dissecting the role and timing of Ag presentation of particular dendritic cell subsets in the initiation of the immune response. The capacity to adoptively transfer small populations of Ag-specific T lymphocytes has also increased our knowledge of the physiologically important role of regulatory T cells in autoimmunity and immunosuppression. New fluorescence imaging techniques such as multicolor video microscopy, laser scanning cytometry, and multiphoton tissue imaging have provided new ways in which researchers can track cellular changes within Ag-specific lymphocytes in vivo. This review summarizes some of the ways in which these techniques have led to discoveries in the role of signaling cascades, cell cycle progression, and apoptosis in maintaining an Ag-specific immune response.
Keywords: adoptive transfer; Ag-specific; cellular interactions; fluorescence; lymphocytes; transgenic
Original Articles
- Structure of the O-polysaccharide of Proteus mirabilis O19 and reclassification of certain Proteus strains that were formerly classified in serogroup O19
Andrei V. Perepelov1, Antoni Różalski2, Beata Bartodziejska2 , Sof’ya N. Senchenkova1 and Yuriy A. Knirel1 (1N.D. Zelinsky Institute of Organic Chemistry, Russian Academy of Sciences, Moscow, Russia. 2 Institute of Microbiology and Immunology, University of Łódź, Łódź, Poland)Abstract. Introduction: Bacteria of the genus Proteus are a common cause of urinary tract infections. The O-polysaccharide chain of their LPS (O-antigen) defines the serological specificity of these bacteria. Based on the immunospecificity of the O-antigens, two species, P. mirabilis and P. vulgaris, were classified into 49 O-serogroups, and more O-serogroups for strains of these species and P. penneri have been subsequently proposed.
Materials and Methods: The lipopolysaccharide of P. mirabilis CCUG 19011 from serogroup O19 was degraded under mildly acidic and mildly alkaline conditions. Polysaccharides thus obtained were studied by chemical methods, including O-deacetylation, sugar and methylation analyses, and 1H- and 13C-NMR spectroscopy. Antisera were obtained by immunization of New Zealand white rabbits with heat-killed bacteria. In serological studies, enzyme immunosorbent assay, passive hemolysis test, and inhibition of passive hemolysis were used.
Results: The following structure of the O-polysaccharide repeating unit was established: →3)-b-D-GlcpNAc-(1→3)-a-D-GalpNAc4,6(R-Pyr)-(1→4)-a-D-GalpA-(1→3)-a-l-Rhap2Ac-(1→ where R-Pyr is (R)-1-carboxyethylidene (an acetal-linked pyruvic acid). This structure is significantly different from the O-polysaccharide structures of P. vulgaris, P. hauseri and P. penneri strains from the same Proteus serogroup O19.
Conclusions: Based on immunochemical studies of the lipopolysaccharides, it is suggested 1) to keep P. vulgaris CCUG 4654 and P. penneri 31 in serogroup O19 as two subgroups, 2) to reclassify P. mirabilis CCUG 19011 into a new Proteus serogroup, O51, and 3) to classify serologically related strains, including P. vulgaris ATCC 49990, P. hauseri 1732-80 and 1086-80, P. penneri 15, and some other P. penneri strains, in yet another Proteus serogroup, O52.Keywords: Proteus mirabilis; lipopolysaccharide; O-polysaccharide; structure; serological classification; pyruvic acid
- Human sera with precipitating antibodies to human soluble immune complexes.A brief communication
Felix Milgrom and Donna Czechowski (Department of Microbiology and Immunology, State University of New York, Buffalo, NY, USA)Abstract.
Introduction: Previous numerous papers by the senior author dealt with the human serum factor referred to as antiantibody which is specifically directed against IgG antibodies that underwent molecular transformation in the course of the reactions with their corresponding antigens. The reactions of this serum factor could be conveniently detected by means of agglutination of Rh-positive erythrocytes sensitized by anti Rh antibodies. No precipitation tests could be developed.
Materials: Most studies were conducted by means of double diffusion in gel precipitation.
Methods and Results: A rheumatoid arthritis serum, G, was noted that produced a strong reaction of double diffusion in gel precipitation with serum samples of a renal graft recipient, T. Further screening detected one more rheumatoid arthritis serum reacting with T; of 28 sera from renal graft recipients, 6 reacted in a similar way to T, but the reactions were considerably weaker and poorly reproducible. Evidence was presented that the precipitin in the two rheumatoid arthritis sera under study had properties of previously described anti-antibody.
Conclusions: Sera with precipitating anti-antibodies may serve as exquisite reagents for detection of soluble immune complexes in human sera.Keywords: rheumatoid arthritis; renal graft; circulating immune complexes
- Serum concentrations of MCP-1 and RANTES in patients during aortic surgery: the relationship with ischemia-reperfusion
Monika Jedynak1 , Andrzej Siemiątkowski.1 , Marek Gacko2, Barbara Mroczko3 and Jacek Borkowski1 (1Department of Anesthesiology and Intensive Therapy, Medical University, Białystok, Poland. 2Department of Vascular Surgery and Transplantology, Medical University, Białystok, Poland. 3Department of Biochemical Diagnostics, Medical University, Białystok, Poland)Abstract.
Introduction: Surgical trauma is associated with depression of the immune system, which results in a high complication rate following abdominal aortic aneurysm (AAA) repair. Monocyte chemotactic protein-1 (MCP-1) and regulated-on-activation normal T cell expressed and secreted (RANTES) protein are important mediators of the immune and inflammatory response. The aim of this study was to determine whether there is any relationship between MCP-1 or RANTES and operative injury and ischemia-reperfusion during AAA surgery in human.
Materials and Methods: Peripheral blood samples were taken from 12 patients before surgery, after anesthesiainduction, before unclamping of aorta (PreXoff), 90 min after unclamping (90minXoff),and at 24 and 48 h after surgery.
Results: The MCP-1 and RANTES serum concentrations were measured with the ELISA technique. MCP-1 concentration significantly increased after reperfusion (90minXoff) in comparison with the PreXoff level (p=0.001). Twenty-four hours aft er AAA repair, MCP-1 significantly decreased 269–225 pg/ml (p=0.005) and reached preoperative value. RANTES level was higher in AAA patients before surgery than in controls (p=0.025) and decreased significantly after ischemia-reperfusion to 13 ng/ml (p<0.001) at 90minXoff. We showed increases in RANTES concentration to 26 ng/ml on the 1st and to 31 ng/ml on the 2nd day after surgery (p=0.020, p=0.012, respectively) compared with the 90minXoff level.
Conclusions: Ischemia-reperfusion during AAA repair results in an increase in MCP-1 and decrease in RANTES concentrations in serum. The changes in chemokine concentrations may influence the development of immunosuppression after AAA repair, contributing to the postoperative course.Keywords: aorta; aneurysm; chemokines; ischemia-reperfusion
- Evaluation of selected peripheral blood leukocyte functions in patients with various forms of periodontal disease
Małgorzata Pietruska1, Janusz Żak2, Jolanta Wysocka2, Barbara Jaworowska2, Beata Żelazowska2 , Alina Lipska2 and Wanda Stokowska1 (1Department of Conservative Dentistry, Medical Academy, Białystok, Poland. 2Department of Pediatric Laboratory Diagnostics, Medical Academy, Białystok, Poland)
Abstract. introduction: Periodontitis (P) is an infectious disease that develops in the supporting tissues of the tooth. One of the risk factors leading to it may be dysfunction of some immune system cells. Therefore, the object of the study was to assess selected functions of peripheral blood leukocytes in patients with various forms of P. As leukocytes are able to secrete interleukin (IL)-4 and IL-6, concentrations of their soluble receptors and the expression of their membrane receptors were investigated.
Materials and Methods: Twenty generally healthy subjects with agressive (AP) and chronic periodontitis (CP) were enrolled in the study. The control group consisted of 8 healthy subjects, with no changes in periodontium. Peripheral blood mononuclear cells (PBMCs) were isolated and cultured. Levels of IL-4, IL-6, and their soluble receptors sIL-4R and sIL-6R were determined in the supernatant by ELISA. The expressions of cell surface IL-4R and IL-6R were assayed on PBMC using flow cytometry.
Results: No statistically significant differences were found in the selected parameters between people with periodontal disease and healthy controls. However, in subjects with AP, there was an increasing tendency in IL-6 concentration and IL-4R expression on PBMCs.
Conclusions: Our results show that leukocytes play a significant part in P and their activity is probably lesion-dependent. Estimation of the cytokines secreted by leukocytes may facilitate differentiation and prognosis of the disease progression.
Keywords: periodontitis; IL-4; IL-6; receptors