Vol. 50, No. 1, 2002
CONTENTS
Review
- Effective Tumor Immunotherapy: Start the Engine, Release the Brakes, Step on the Gas, …and Get Ready to Face Autoimmunity
Inigo Tirapu, Guillermo Mazzolini, Mercedes Rodriguez-Calvillo, Ainhoa Arina, Belen Palencia, Izaskun Gabari and Ignacio Melero (Gene Therapy Division, Facultad de Medicina and Clinica Universitaria, Universidad de Navarra, Pamplona, Spain)Abstract. Cellular immune responses can destroy cancer cells, achieving the cure of experimental malignancies. An expanding wealth of knowledge on the molecular basis of how to prime and amplify a T cell response has fueled a number of strategies successful at treating established tumors (rather than merely preventing tumor grafting). The most efficacious approaches operate at different stages, including: 1) priming the immune response using tumor antigen-expressing dendritic cells or tumor cells transfected with genes that render them immunogenic, 2) sustaining and amplifying immunity using agonistic monoclonal antibodies against costimulatory molecules or immune-potentiating cytokines, and 3) eliminating mechanisms that self-regulate the strength of the immune response, such as inhibitory receptors or regulatory T cells. A rational combination of such approaches holds great hope for cumulative and synergistic effects, but there is also evidence that they can open the flood-gates for unwanted inflammatory reactions. The next decade can be envisioned as the time when the first reproducibly efficacious combination regimes for cancer immunotherapy will become available and widely used in the clinic, as clinicians learn the best strategies and try to harness their potentially damaging effects.
Keywords: immunotherapy; dendritic cells; costimulation; CTLA-4; 4-1BB; CD40; cytokines.
- Caspases – Their Role in Apoptosis and Other Physiological Processes as Revealed by Knock-Out Studies
Kenneth Sadowski-Debbing (Clinic for Craniomaxillofacial Surgery, Schloss Str. 1, D-48683 Ahaus, Germany), Johannes F. Coy (MTM Laboratories AG, Im Neuenheimer Feld 519, D-69120 Heidelberg, Germany), Walter Mier (Institute of Nuclear Medicine, University of Heidelberg, D-69120 Heidelberg, Germany), Hubert Hug (University Childrens Hospital, Prittwitzstr. 43, D-89075 Ulm, Germany), Marek Los (Department of Immunology and Cell Biology, University of Muenster, D-48149 Muenster, Germany)Abstract. Caspases are crucial mediators of apoptosis, a form of physiological cell death. Their activation is carefully controlled by a philogenetically conserved death program, which is indispensable for the homeostasis and development of higher organisms. Dysregulation of apoptosis contributes to the pathogenesis of many human diseases. As effectors of the apoptotic machinery, caspases are considered potential therapeutic targets. In vitro studies have demonstrated the requirement of caspases activity for both the triggering phase as well as the execution of apoptosis, thus providing a molecular base for the fine-tuning of this process by pharmacological agents. The precise roles of the individual caspases in vivo and their functional relation to each other have been best demonstrated in genetically modified animals. The generation of single caspase-deficient mice have confirmed most of the data obtained in vitro and exposed some new aspects previously undetected in the cell culture system. Interestingly, inactivation of many caspases revealed not only their expected participation in apoptotic events as well as in the maturation of cytokine, but also provided hints about the role of at least some caspases in cell differentation and stimulatory repsonses. In this review we will discuss what these studies have unveiled about the role of individual caspases in development, apoptosis, and inflammation, with particular focus on their role beyond the apoptotic process.
Keywords: caspases; cell death; development; cell cycle; inflammation.
- Emerging Concepts in the Molecular Pathogenesis of Systemic Lupus Erythematosus
Madhusoodana P. Nambiar, Yuang-Tuang Juang and George C. Tsokos (Department of Cellular Injury, Walter Reed Army Institute of Research, Building 503, Robert Grant Avenue, Silver Spring, MD 20910-7500, USA, Department of Medicine, Uniformed Services University of the Health Sciences, Bethesda, MD 20814, USA)Abstract. Systemic lupus erythematosus is a prototypic autoimmune disease that predominantly afflicts women during child-bearing age. The disease is characterized by the production of autoantibodies and immune complexes in association with a diverse array of clinical manifestations. Investigation into the etiopathogenesis has been directed at identifying the genes that provide susceptibility to the disease, the complex cellular and cytokine aberrations and the biochemical abnormalities that are responsible for them. Understanding the immune cell signaling and gene transcription abnormalities will help us tailor new strategies for efficient biotherapy of the disease.
Keywords: systemic lupus erythematosus; autoimmune diseases; pathogenesis.
- Interferon Tau and Its Immunobiological Role in Ruminant Reproduction
Anna Chełmońska-Soyta (Department of Veterinary Prevention and Immunology, Faculty of Veterinary Medicine, Agriculture University of Wrocław, Norwida 31, 50-375 Wrocław, Poland)Abstract. Interferon tau (IFN-tau) is an key cytokine in maintaining pregnancy in ruminants. It is produced by the ruminant conceptus around the time of implantation. IFN-tau belongs to the type I interferon family but, unlike the other members of this group, it is not virus inducible and its expression is temporal and restricted to the trophoblast cells of the ruminant conceptus. The main target of the paracrine action of this cytokine is the endometrium. It changes the prostaglandin metabolism and secretory function of the cells by upregulating the secretion of several proteins. It also presents immunomodulatory action towards leukocytes by changing their proliferative responses and cytokine production. This cytokine activity in reproductive biology and immunology has been intensively explored for the last ten years. It has been regarded as a potential tool in improving the performance and biotechnological processes in ruminant reproduction. Additionally, its high antiviral potency and low cytotoxicity in comparison with IFN-tau has placed this cytokine in the group of possible therapeutics in human and animal medicine.
Keywords: interferon tau; cytokine; ruminants; reproduction; pregnancy.
- The Role of the Innate and Adaptive Immune Responses in Acanthamoeba Keratitis
Jerry Y. Niederkorn (Department of Ophthalmology, University of Texas Southwestern Medical Center Dallas, Texas 75390-9057, USA)Abstract. Infections of the corneal surface are an important cause of blindness. Protozoal, viral, bacterial, and helminthic infections of the cornea account for up to nine million cases of corneal blindness. Free-living amoebae of the genus Acanthamoeba produce a progressive infection of the cornea called Acanthamoeba keratitis. Disease is usually transmitted by Acanthamoeba trophozoites bound to soft contact lenses. Infection of the cornea is initiated when the parasite binds to the corneal epithelial surface. Recrudescence can occur and suggests that the adaptive immune response is not aroused by corneal Acanthamoeba infections. Systemic immunization with Acanthamoeba antigens elicits robust Th1 cell-mediated immunity and serum IgG antibody, yet fails to prevent the development of Acanthamoeba keratitis. However, immunization via mucosal surfaces induces anti-Acanthamoeba IgA antibodies in the tears and provides solid protection against the development of Acanthamoeba keratitis. Unlike other immune effector mechanisms that rely on cytolysis, inflammation, release of toxic molecules, or the induction of host cell death, the adaptive immune apparatus prevents Acanthamoeba infections of the cornea by simply preventing the attachment of the parasite to the epithelial surface. The beauty of this mechanism lies in its exquisite simplicity and efficacy.
Keywords: cornea; Acanthamoeba keratitis; corneal blindness.
Immunochemistry
- Thermal Glycation of Proteins by D-Glucose and D-Fructose
Urszula Kańska and Janusz Boratyński (Department of Tumor Immunology, Institute of Immunology and Experimental Therapy, Polish Academy of Sciences, 12 Weigla, 53-114 Wroclaw, Poland)Abstract. The dry thermal glycation method was used to conjugate D-glucose and D-fructose with bovine serum albumin. Reactions were conducted at 50-104oC for 30 min. Depending on temperature, different levels of substitution were achieved. In the case of D-glucose average substitution level of up to 53 mol glucose/mol BSA were obtained. D-Fructose turned out to be less reactive than D-glucose. The pH of the reaction mixture was also found to affect the efficiency of the glycation reaction. The levels of substitution were estimated using matrix assisted laser desorption/ionisation time of flight mass spectrometry (MALDI-TOF) and SDS-polyacrylamide gel electrophoresis. Glycated molecules, and a process of glycation itself are implicated in diabetes complications. Hypothetically bovine spongiform encephalopathy (BSE) was also derived from glycation.
Keywords: glycation; BSA; D-glucose; D-fructose; conjugate.
- Glycoforms of Six Serum Glycoproteins in a Patient with Congenital Disorder of Glycosylation Type I
Mirosława Ferens-Sieczkowska, Iwona Kątnik-Prastowska (Deptartment of Chemistry and Immunochemistry, Wrocław Medical University, Bujwida 44A, 50-345 Wrocław, Poland), Krzysztof Zwierz (Department of Pharmaceutical Biochemistry), Alina Midro (Department of Clinical Genetics, Medical Academy, Mickiewicza 2A, 15-222 Białystok, Poland)Abstract. In this paper the occurrence and relative content of defectively glycosylated serum glycoforms in transferrin (Tf), a1-acid glycoprotein (AGP), haptoglobin (Hp), a1-antitrypsin (a1-AT), a2-macroglobulin (a2-MG) and ceruloplasmin (Cpl) in the serum of a patient with congenital disorder of glycosylation type I are reported. Blood samples were taken when the patient was 14 years old and then after a one-year interval. The patterns of glycoforms in both samples were compared. In 4 out of 6 examined glycoproteins, glycoforms lacking one and two oligosaccharide chains occurred. „Underglycosylated” glycoforms of a2-MG and Cpl were not clearly detectable. Tf was shown to be affected with this defect to a higher extent than other glycoproteins, containing only 30% properly glycosylated molecules and also as much as 30% of the molecules lacking two glycan units. In Hp and a1-AT the proportions of properly and defectively glycosylated forms were similar. This properly glycosylated form comprised 47% of the Hp and 51–55% of the a1-AT molecules. As in AGP and Tf, about 30% the of molecules lacked one glycan unit. Twenty-one percent of the Hp molecules were devoid of two glycans, and this amount slightly increased in the course of the year. In a1-AT, 19 and 17% of the molecules lacked two glycans in both samples, respectively. Only in AGP we did find a substantial difference between the two blood samples. In the course of the year, the amount of the form lacking 2-chains decreased from 12 to 3%, resulting in a simultaneous increase in the forms lacking one chain and the properly glycosylated. Our work also indicates, that applying a simple method of biochemical analysis such as SDS-PAGE/Western-blotting could be helpful in preliminary diagnosis and could improve the identification of congenital disorders of glycosylation.
Keywords: glycosylation; congenital disorders of glycosylation; a1-acid glycoprotein; a1-antitrypsin; haptoglobin; transferrin; a2-macroglobulin; ceruloplasmin.
Clinical Immunology
- Comparison of Leukocyte Populations from Bronchoalveolar Lavage and Induced Sputum in Evaluation of Cell Composition and Nitric Oxide Production in Patients with Bronchial Asthma
Małgorzata Bieńkowska-Haba, onika Cembrzyńska-Nowak, Iwona Siemieniec (Laboratory of Virology, Institute of Immunology and Experimental Therapy, Polish Academy of Sciences, Weigla 12, 53-114 Wrocław, Poland), Jerzy Liebhart, Rafał Dobek, Ewa Liebhart, Bernard Panaszek, Andrzej Obojski, Józef Małolepszy (Department of Internal Medicine and Allergology, Wrocław Medical University, Traugutta 57/59, 50-454 Wrocław, Poland)
Abstract. Bronchoalveolar lavage (BAL) or induced sputum (IS) techniques may provide leukocytes for the evaluation of airway inflammatory response in bronchial asthma. The aim of the present study was to compare features of leukocyte populations obtained by the two different methods regarding the cell types and their activity in patients with bronchial asthma. The nitric oxide (NO) level released from the cells was measured as a marker of their activity. Pulmonary leukocytes were obtained from the BAL and IS of 11 asthmatic patients in stable condition at the time of the study. The BAL and IS leukocyte populations varied in cell count and NO production. Macrophages were the predominant leukocyte population in BAL (Me = 83.0%, range 67.9-88.4%), whereas sputum sediments were found to consist mainly of neutrophils (Me = 55.7%, range 29.0-64.9%). The IS leukocytes released much more NO (p = 0.0022) than the BAL leukocytes. In spite of these quantitative differences, a similar pattern of NO production was observed in BAL and in IS cells. Both BAL and IS leukocyte populations produced almost the same amounts of NO before and after lipopolysaccharide stimulation (p = 0.9063, p = 0.4801, respectively). Furthermore, a slight positive correlation (RS = 0.5578, p=0.0594) was noticed between the neutrophil percentages and NO levels produced by BAL cells, whereas in IS a statistically significant correlation between the percentage of neutrophils and the levels of NO (RS = 0.6643, p = 0.0184) was observed. In conclusion, the BAL and IS leukocyte populations are different in cell type, their size and activity. Depending on the asthma severity and the type of cells needed in a study, either BAL or IS specimens may be chosen as a source of pulmonary leukocytes. The use of IS as a noninvasive technique is supposed to be potential value particularly in the study of the airway inflammatory response mediated mainly by neutrophils, i.e. during and/or after exacerbation of the disease. Based on our results, a possible contribution of neutrophils in the production of NO in the airways of asthmatic patients can be proposed apart from other cells such as macrophages.
Keywords: bronchoalveolar lavage, induced sputum, pulmonary leukocytes, nitric oxide, neutrophils, macrophages, eosinophils, bronchial asthma
Vol. 50, No. 2, 2002
CONTENTS
Review
- CD45 in Memory and Disease
Elma Z. Tchilian and Peter C.L. Beverley (The Edward Jenner Institute for Vaccine Research, Compton, UK)Abstract. CD45 (the leukocyte common antigen) is known to function as a tyrosine phosphatase in leucocyte signalling. Biochemical studies indicate that CD45 is involved in the regulation both of T cell receptor-associated kinases and Janus kinases that transmit signals from cytokine receptors. However the function of the different isoforms of CD45 generated by complex alternative splicing, and indeed the role of the whole extracellular domain of the molecule, remain mysterious. Analysis of CD45 knockouts and of transgenic mice expressing single CD45 isoforms, as well as the disease associations of human polymorphisms, is providing new insights into CD45 function. Accumulating data from these genetic and biochemical studies promises to elucidate the role of high and low molecular weight isoforms of CD45 in the function of naive and memory T lymphocytes.
Keywords: CD45; cellular signaling; memory T lymphocytes.
- Low Molecular Weight Protein Tyrosine Phosphatase and Human Diseases: in Search of Biochemical Mechanisms
Nunzio Bottini, Tomas Mustelin (Program of Signal Transduction, La Jolla Cancer Research Center, The Burnham Institute, 10901 North Torrey Pines Road, La Jolla, CA 92037, USA), Egidio Bottini (Department of Neurosciences), Fulvia Gloria-Bottini (Department of Biopathology, University of Rome Tor Vergata, Rome, Italy)Abstract. A major challenge in the post-genomic era is to identify the physiological function of genes and elucidate the molecular basis for human disease. Genetic polymorphisms offer a convenient aveune for these efforts by providing evidence for the involvement of a given gene in human pathophysiology. Here we review the current evidence linking the low molecular weight protein tyrosine phosphatase (LMPTP) to several common diseases, including allergy, asthma, obesity, myocardial hypertrophy, and Alzheimer’s disease. Based on the know effects of the genetic polymorphisms on the alternative mRNA splicing and enzyme levels of LMPTP, we discuss the possible molecular mechanisms of LMPTP involvement in these diseases.
Keywords: tyrosine phosphate; genetic polymorphisms; allergy; asthma; obesity.
- Significance of Tumor-Cell Receptors in Human Cancer
Jan Żeromski (Chair and Department of Clinical Immunology, Karol Marcinkowski University of Medical Sciences, Poznań, Poland)Abstract. Every tumor cell is equipped with an array of biologically active surface molecules, and several these function as receptors for various ligands. They include MHC, or in the case of humans, HLA antigens, cytokine receptors, cell-adhesion molecules, growth factor receptors, Fas/Fas-ligand molecules and others. Their expressions are a subject to alterations, usually to the advantage of tumor growth and spread. Some appear on tumor cells de novo, having no counterparts on the respective normal cells. Detailed knowledge about the expression of tumor-cell receptors and their genotypes, in particular of cancerous ones, may provide information essential for the creation of tools for specific tumor immunotherapy.
Keywords: HLA antigens; cytokine receptors; adhesion molecules; growth factors.
- Potential for Immune Reconstitution by G-CSF Treatment in HIV Patients
Sonja von Aulock and Thomas Hartung (Biochemical Pharmacology, University of Konstanz, Konstanz, Germany)Abstract. New treatment strategies for HIV/AIDS are very successful in reducing viral load. However, reconstitution of the immune system takes about one year and may be insufficient or remain incomplete. During this time the patient remains prone to opportunistic infections as a result of the complex immune dysfunction caused by the virus. Recombinant granulocyte colony-stimulating factor (G-CSF) has diverse immunomodulatory properties which may be beneficial to aid immune reconstitution.
Keywords: HIV; AIDS; G-CSF; immune reconstitution.
- GM-CSF Regulation in Eosinophils
Stéphane Esnault and James S. Malter (Department of Pathology and Laboratory Medicine, University of Wisconsin, Madison, WI, USA)Abstract. Allergic asthma is characterized by a temporally and quantitatively inappropriate immunologic response. One of hallmarks of this response is the accumulation of eosinophils in the airway and lung parenchyma, which results in bronco-constriction, lung damage and ultimately fibrosis. GM-CSF plays a pivotal role in this process by modulating eosinophil function and survival. In this review, we discuss the effects and molecular regulation of GM-CSF secretion by eosinophils. Recent data demonstrate activated eosinophils release small amounts of anti-apoptotic GM-CSF by stabilizing its coding mRNA.
Keywords: eosinophils; GM-CSF; mRNA; survival; asthma.
- Immunomodulation of Macrophages by Pathogenic Yersinia Species
Klaus Ruckdeschel (Max von Pettenkofer-Institute for Hygiene and Medical Microbiology Pettenkoferstr. 9a, 80336 Munich, Germany)Abstract. The interaction between macrophages and bacterial pathogens plays a crucial role in the pathogenesis of infectious diseases. Pathogenic species of the gram-negative bacterium Yersinia deploy complex strategies to disarm macrophages and to disrupt their response to infection. For this purpose, Yersinia spp. engage a type III protein secretion system that mediates polarized translocation of Yersinia virulence factors, the so-called Yops, into the host cell cytoplasm. There, the Yops act on different cellular levels to neutralize a sequence of programmed phagocyte effector functions. Yersiniae initially impair the phagocytic machinery and block the generation of the bactericidal oxidative burst. Furthermore, yersiniae uncouple an array of fine-tuned signals of innate immunity, which leads to suppression of the macrophage TNF-a production and to macrophage apoptosis. The impairment of cellular functions results in a scenario, by which Yersinia efficiently resist the attack of the macrophage and finally kills the macrophage by activating its intrinsic cell suicide mechanism. This review highlights the aspects of Yersinia-macrophage interaction that determine the fate of the infected cell.
Keywords: Yersinia; macrophage; apoptosis; YopP/YopJ; NF-kB.
Clinical Immunology
- Effect of IL-18 on IL-lb and sIL-1RII Production by Human Neutrophils
Ewa Jabłońska, Aneta Iżycka and Natalia Wawrusiewicz (Department of Immunology, Medical Academy of Białystok, Poland)Abstract. In the present study we investigated the effect of interleukin-18 (IL-18) on the production of interleukin-1b (IL-1b) and soluble interleukin-1 receptor II (sIL-lRII) by human neutrophils. The results obtained indicate that recombinant human interleukin-18 (rhIL-18) induces IL-lb and, to a lesser extent, sIL-1RII production by human neutrophil isolated from peripheral blood. However, this effect was less important than lipopolysaccharide (LPS) stimulation. Additionally, our observations suggest that IL-18 can induce priming of neutrophils for IL-lb and, to a lesser extent, sIL-lRII production by LPS-stimulated cells. The ability of IL-18 to serve as an effective modulator for IL-lb and its regulatory protein may have significance in the inflammatory and immune reactions mediated by IL-lb.
Keywords: neutrophil, interleukin-18, interleukin-1b, soluble interleukin-1 receptor II.
- A Study of Thyroglobulin Concentration in the Thyroid and Serum of Patients with Different Thyroid Disorders
Agata Czarnywojtek, Iwona Krysińska, Katarzyna Łącka, Maciej Gembicki (Department of Endocrinology, Institute of Internal Medicine, K. Marcinkowski University of Medical Sciences, Przybyszewskiego 49, 60-355 Poznań, Poland), Bolesław Stawny (Department of General and Gastroenterological Surgery, Poznań, Poland), Michał Rólski (Department of Children Surgery, University of Medical Sciences, Poznań, Poland), Jan Włoch (Oncology Centre, M. Skłodowska-Curie Institute, Gliwice, Poland)
Abstract. Knowledge concerning the structure and quality of thyroglobulin (Tg) has great significance for the better understanding of the pathogenesis of different thyroid diseases. The localisation of the Tg gene and studies of its structure by molecular biological techniques make possible precise investigations of its expression. The aim of our study was to evaluate Tg content in the thyroids and Tg concentrations in the serum of 108 patients suffering from benign or malignant thyroid disorders. The method of investigation was isolating total protein from thyroid tissues obtained during surgery and determining Tg content in the thyroid extracts and Tg concentrations in serum. The Tg concentrations in serum and in thyroid protein extracts were evaluated by fluoroimmunometric assay. Statistical analysis was carried out with the help of the computing programmes.
Keywords: thyroglobulin; thyroid disorders; protein extracts.
Vol. 50, No. 3, 2002
CONTENTS
Review
- Components of the IFN-g Signaling Pathway in Tumorigenesis
George Blanck (Department of Biochemistry and Molecular Biology, Moffitt Cancer Center, University of South Florida College of Medicine, Tampa, FL 33612, USA)Abstract. Many features of the interferon g (IFN-g) signaling pathway would suggest that it is anti-tumorigenic. The IFN-g signaling pathway leads to apoptosis and to the expression of immune function proteins that could cooperate with T cells in the destruction of tumor. Various lines of experimental approaches have in general supported the hypothesis the IFN-g signaling pathway is anti-tumorigenic. However, data also indicate that the idea that the IFN-g signaling pathway is exclusively anti-tumorigenic is too simplistic. Also, to date, very little of the knowledge regarding the anti-tumor effects of the IFN-g pathway has been useful in the prognosis or therapy for cancer. This review summarizes the current state of knowledge regarding the IFN-g signaling pathway in tumorigenesis, with an emphasis on MHC class II induction in tumor cells and the induction of apoptosis in tumor cells. The review also indicates some future areas of investigation that offer hope for applying this knowledge in reducing cancer mortality.
Keywords: interferon gamma; tumorigenesis; apoptosis; MHC; interferon regulatory factor-1; retinoblstoma protein.
- Modulation of Pulmonary Innate Immunity during Bacterial Infection: Animal Studies
Marcus J. Schultz1,2 and Tom van der Poll1,3 (1Laboratory of Experimental Internal Medicine, 2Department of Intensive Care Medicine, 3Department of Infectious Diseases, Tropical Medicine and AIDS, Academic Medical Center, University of Amsterdam, Amsterdam, Netherlands)Abstract. Both the increasing number of immunocompromised patients susceptible to pneumonia, and the development of bacterial resistance are significant problems related to the treatment of pneumonia. The primary outcome of treatment for pneumonia is to tip the balance to a successful host response. An ideal approach would be the combination of immunomodulation and conventional antimicrobial therapy for the treatment of pneumonia. It is of increasing importance to understand the components of innate immunity, before immunomodulatory therapy can be applied to patients. Much of our knowledge of the role of alveolar macrophages, cytokines and chemokines in the pathogenesis of pneumonia is derived from animal studies on experimental pneumonia. This article summarizes current information on the role of an alveolar macrophage (AM) and AM-derived mediators in host defense against pneumonia.
Keywords: innate immunity; pneumonia; alveolar macrophages; cytokines; immunotherapy.
- CD28 Costimulatory Molecule – Expression, Structure and Function
Dorota Boćko1, Agata Kosmaczewska1, Lidia Ciszak1, Renata Teodorowska1 and Irena Frydecka1,2 (1Institute of Immunology and Experimental Therapy, Polish Academy of Sciences, Wrocław, Poland, 2Department of Hematology, Medical University, Wrocław, Poland)Abstract. T cell activation is a key event triggering an antigen specific immune response of the organism. The process is induced primarily by signal generated by direct interaction of T cell receptor (TCR) with antigen bound to major histocompatibile complex (MHC) on antigen presenting cell (APC). Although the signal is critical to excite immune response, however additional, costimulating signal is required. The major second signal is generated by interaction of CD28 molecule expressed on most T lymphocytes with its natural ligands CD80 and CD86 located on APCs. Signal excited by CD28 triggering involves multiple second messenger cascades, leading to activation of transcription factors and finally results in cell proliferation, cytokine production, and generation of effector function. The importance of CD28-delivered costimulatory signals was proven in experiments with CD28-deficient mice. T cells from these mice exhibit, impaired pattern of cytokine secretion, defects in T cell dependent antibody production. Certain forms of immunopathology might result from the aberrant regulation of CD28 expression.
Keywords: T cell activation; CD28 molecule; costimulation.
- Mast Cells and Inflammation
Michael Stassen1, Lothar Hültner2, Christian Müller1 and Edgar Schmitt1 (1Institute of Immunology, Johannes Gutenberg-University, Mainz, Germany, 2GSF-National Research Center for Environment and Health. Institute of Clinical, Molecular Biology and Tumor Genetics, Munich, Germany)Abstract. Mast cells have long since been recognized as potent producers of a large panel of biological highly active mediators such as biogenic amines, arachidonic acid metabolites, cytokines and chemokines, but most of their biological functions had been elusive and speculative. By taking advantage of mast cell-deficient mice, the role of mast cells in a variety of experimental settings can now be studied in detail and such approaches have dramatically altered and enlarged our knowledge about mast cell biology and function. Herein we will focus on the role of mast cells in inflammatory reactions of diverse origin such as delayed type hypersensitivity, atopy, immune complex-mediated inflammation and innate immune responses. From a current point of view, there is no doubt that the most outstanding and beneficial feature of mast cells is their recently uncovered ability to rapidly induce a life-saving inflammatory response upon encountering microbes and microbial constituents. Nevertheless, the picture is also emerging that mast cells are deeply involved in the induction and maintenance of a variety of severe allergic and autoimmune diseases. However, a deeper understanding of their activation and immune-modulatory capacity might open a new window for the development of curative strategies.
Keywords: mast cells; inflammation; IgE-independent activation; innate immune response.
- The Role of Dendritic Cells in Neurodegenerative Diseases
Pablo Iribarren, You-Hong Cui, Yingying Le and Ji Ming Wang (Laboratory of Molecular Immunoregulation, Center for Cancer Research, National Cancer Institute at Frederick, Frederick, MD 21702, USA.)Abstract. Dendritic cells (DCs) are the most potent antigen-presenting cells (APCs) involved in the induction of adaptive immune responses. The presence of DCs in the central nervous system (CNS) and the active participation of the immune system in a variety of neurodegenerative diseases have been demonstrated. This review will discuss recent findings pertinent to DCs and other antigen presenting cells in CNS in health and disease states.
Keywords: dendritic cells; neurodegenerative diseases; CNS.
- Active Vaccination after Allogeneic Bone Marrow Cell Transplantation: a New Option in the Immunotherapy Cancer?
Margot Zöller1, 2 and Siegfried Matzku2, 3 (1Department of Tumor Progression and Immune Defense, German Cancer Research Center, Heidelberg, Germany, 2Department of Applied Genetics, University of Karlsruhe, Karlsruhe, Germany, 3Department of Oncology, Biomedical Research, Merck KGaA, Darmstadt, Germany)Abstract. The concept of immunotherapy of cancer has been evoked more than a century ago by W. Coley. Yet, it is only recently that the state of knowledge allows for molecularly defined therapeutic approaches and much effort will still be required to place immunotherapy beside of surgery, chemotherapy and radiation as a forth option. In this review, we will strongly focus on two aspects: active therapeutic vaccination, because it is our belief that this approach will provide a major breakthrough and the potential efficacy of combining active vaccination with allogeneic bone marrow cell transplantation. It lately could be established in clinical trials that allogeneic bone marrow cell transplantation does not require myeloablative conditioning. Only non-myeloabaltive conditioning, which avoids the high toxicity of the conventional approach, allows the recruitment of elderly patients and patients in poor health condition. Concerning active vaccination protocols we will address the questions 1) what the targets (i.e. the antigens) of immunotherapeutic approaches could be; 2) how to achieve an optimal confrontation of the immune system with these tumor-associated antigens; and 3) which response elements are needed for raising a therapeutically successful immune reaction against these. Many question remain to be answered in the field of allogeneic bone marrow transplantation after non-myelablative conditioning to optimize the therapeutic setting for this likely very powerfull tool of cancer therapy. We will briefly summarize current considerations to improve engraftment, to reduce graft versus host disease while strengthening graft versus tumor reactivity. There is some hope that the latter can be „naturally” maintained during the process of T cell maturation in the allogeneic host. Provided this hypothesis can be substantiated, the efficacy of active vaccination of the allogeneically reconstituted host will meet a pool of virgin T cells, which are tolerant towards the host, but not anergized towards tumor antigens presented by MHC molecules of the host. We only briefly will mention suportive regimen of immunomodulation and those hazards which one is most frequently confronted with in trials to attack tumors with the inherent weapon of immune defense. Though successful immunotherapy of cancer still remains far behind expectation, there is a solid basis to believe that by improving our understanding of molecular mechanisms of immunity, it may become a very powerful and less harmful tool than conventional therapies.
Keywords: tumor, vaccination, allogeneic, stem cell transplantation
Experimental Immunology
- A Two Step Procedure to Fractionate Mouse Testicular Macrophages with a Different Cytokine Profile
Krzysztof Bryniarski, Katarzyna Szewczyk, Maria Ptak, Małgorzata Bobek and Włodzimierz Ptak (Department of Immunology, College of Medicine, Jagiellonian University, Czysta 18, 31-121 Kraków, Poland)
Abstract. Cells isolated enzymatically from interstitial tissue of mouse male gonads are composed of macrophages, Leydig cells, and myofibroblasts. They can be separated on density gradients either by sedimentation (Ficoll) or flotation (Percoll) into several fractions according to different buoyant density containing mixtures of different cells. Macrophages (FcgR+, esterase+) present in cell mixtures can by highly enriched in a single step to 95% purity by rosetting with opsonized erythrocytes followed by sedimentation on Lymphoprep. Separate fractions of highly purified (over 95%) macrophages obtained by successive use of density gradients and rosetting differ significantly in the production of cytokines, such as cells from fractions at lower density produce little IL-6, cells from fractions at higher density are poor producers of TNF-alpha whereas TMf in intermediate fractions produce significant amounts of both cytokines. These differences may suggest that particular subpopulations of testicular macrophages play different biological roles in the testis.
Keywords: testicular macrophages; isolation; functional subpopulations; cytokine secretion; FcgR expression.
Vol. 50, No. 4, 2002
CONTENTS
Review
- Allergen-Specific T Lymphocytes as Targets for Specific Immunotherapy: Striking at the Roots of Type I Allergy
Barbara Bohle (Department of Pathophysiology, Division of Immunopathology, University of Vienna, Vienna, Austria)Abstract. In the past decades allergic diseases have tremendously increased and hypersensitivity reactions now represent a growing health concern in industrialized countries. Despite various effective therapeutic options for the treatment of allergic diseases, only specific immunotherapy (SIT) has been shown to have effects on the underlying immunological mechanisms, namely functional changes at the level of T helper lymphocytes (Th). It was found that allergen-specific CD4+ Th2 lymphocytes play a key role in the pathophysiology of atopic diseases. During successful SIT, the Th2-dominated immune response is modified towards a Th1 response, leading to a decline in allergen-specific IgE levels in the long term. In order to improve the efficacy and safety of SIT, novel approaches were developed targeting allergen-specific Th2 lymphocytes since specific inactivation or modulation towards Th1 cells could interfere with the disease process. In view of this aspect, this review will basically focus on two new promising approaches to improve SIT: 1) the use of hypoallergenic proteins characterized by reduced IgE-binding capacities but retained T lymphocyte-activating properties and 2) oligodeoxynucleotides containing CpG motifs as an example of adjuvants which foster Th1 immune responses. Both approaches promise to be capable of adjusting the pathological Th2 immune response.
Keywords: type I allergy, specific immunotherapy, T lymphocytes, hypoallergens, oligodeoxynucleotides containing CpG motifs.
- Type I Interferons as Immunoregulatory Molecules; Implications for Therapy in Experimental Autoimmune Uveoretinitis
Junichiro Mizuguchi1, Masaru Takeuchi2 and Masahiko Usui2 (1Department of Immunology, Tokyo Medical University, 6-1-1 Shinjuku, Shinjuku-ku, Tokyo 160-8402 Japan, 2Department of Ophthalmology, Tokyo Medical University Hospital, 6-7-1 Nishishijuku, Shinjuku-ku, Tokyo, 160-0023 Japan)Abstract. Clinical trials have shown that the type I interferon (IFN)-alpha/beta have some beneficial effects on organ-specific autoimmune disease, such as Behcet’s diseases and multiple sclerosis, although the precise mechanims remain lergaly unresolved T helper cells (Th1)-mediated autoimmune response are involved in the initiation and/or progression of human uveitis, such as Behcet’s disease. The animal model of experimental autoimmune uveoretinitis (EAU), characterized by a monophasic clinical course, has contributed to the understanding of the pathogenesis of human uveitis. Th1 producing IFN-gamma induce EAU development, while Th2 producing IL-4/IL-10 prevent the disease. However, depending on the cytokine milieu, the pro-inflammatory cytokine IFN-gamma may attenuate the autoimmune response and anti-inflammatory cytokine IL-4 exacerbates it. Chemokines also play a crucial role in EAU development, which might be resolved by Th2-mediated immune responses. The administration of IFN-alpha/beta prevents EAU development, accompanied by a dimished production of IFN-gamma/IL-10. Interestingly, however, IFN-alpha/beta also have some beneficial effects on patients with Th2-like phenotype in addition to Th1-like phenotypes. Thus, the immuno-modulatory action of IFN-alpha/beta may be dependent on the context of cytokine combination and/or their concentrations.
Keywords: interferon alpha/beta; autoimmune diseases; T helper cells; human uveitis.
- V Gene Replacement in T and B Lymphocytes: Illicit or Regimented Rearrangement?
Rachel Golub (Unité du Développement des Lymphocytes, Institut Pasteur, 75724 Paris Cedex 15, France)Abstract. Lymphocyte can undergo novel types of secondary genetic rearrangements that alter the specificity of a pre-existing B cell receptor (BCR) or T cell receptor (TCR). V gene replacement is one of them and it replaces an already rearranged V gene segment with an upstream germline V gene segment. This review focuses on the molecular aspect of rearrangement. The role of this mechanism in the immune system is debated.
Keywords: VDJ recombination, immunoglobulin and TCR loci, embedded heptamer, knock-in mice, gene rearrangement, receptor editing.
- The TCR/CD3 Complex: Molecular Interactions in a Changing Structure
Maria José Feito1, Arturo Jiménez-Perianez2, Gloria Ojeda2, Alejandra Sánchez1, Pilar Portolés2 and José M. Rojo1 (1Departmento de Inmunología, Centro de Investigaciones Biológicas, CSIC, Madrid, Spain, 2Centro Nacional de Biología Fundamental, Instituto de Salud Carlos III, Majadahonda, Madrid, Spain)Abstract. The T cell receptor-CD3 (TCR/CD3) complex is a multichain structure in charge of antigen recognition in T cells. Despite many genetic, structural, and functional data obtained in recent years, essential questions concerning the TCR/CD3 complex still remain open, including: 1) the precise number of polypeptides in each TCR/CD3 complex, their interactions and spatial arrangement; 2) the role(s) of each polypeptide in antigen recognition and/or in receptor signal transmission, and 3) the relationship between the TCR/CD3 complex and other membrane or cytoplasmic molecules involved in downstream signaling. In this work we shall review data concerning some of these issues, proposing a model of the overall structure of the TCR/CD3 complex to explain its known features.
Keywords: T cell receptor; CD3; antigen recognition.
- DNA Viruses and Genetic Modification of Dendritic Cells
Lars Jenne and Alexander Steinkasserer (Department of Dermatology, University of Erlangen-Nürnberg, Hartmannstraße 14, D-91052 Erlangen, Germany)Abstract. An alternative approach to the use of tumor-peptide-loaded dendritic cells (DC) in immunotherapy would be the use of genetically modified DC using viral vectors expressing tumor-associated antigens (TAA). However, viruses have developed several immune escape mechanisms and, thus, one has to study the interaction between viruses and DC before these viruses can be used as an alternative strategy. Here we report that vaccinia virus (VV) as well as herpes simplex virus type-1 (HSV-1) are able to potently infect monocyte-derived DC, however, this infection leads to the inhibition of the DC-mediated T cell stimulation in vitro.
Keywords: dendritic cells; viral vectors; gene therapy; vaccinia virus; herpes simplex virus.
- The Emerging Distinct Role of TNF-Receptor 2 (p80) Signaling in Chronic Inflammatory Disorders
Martin H. Holtmann, Marcus Schuchmann, Geraldine Zeller, Peter R. Galle and Markus F. Neurath (Department of Medicine, Johannes-Gutenberg-University, Mainz, Germany)Abstract. Tumor necrosis factor (TNF)-alpha is a pleiotropic cytokine with strong proinflammatory and immunomodulatory properties. TNF-alpha plays a critical role in many acute or chronic inflammatory diseases and anti-TNF strategies have proven to be clinically effective. Two TNF-specific cell surface receptors, TNF-R1 (p60) and TNF-R2 (p80), have been identified and the function of these receptors and the downstream intracellular signal-transduction pathways have been extensively studied in vitro. For a long time p60 was considered to be the predominant mediator of TNF signaling, whereas p80 was ascribed only an euxilliary function. However, there is increasing clinica and experimental evidence for an important independent role of p80 signaling in chronic inflammatory conditions. The date, most data exist for Crohn’s disease. Upregulation of p80 and increased p80 signaling aggrevates experimental colitis and is likely to contribute to the chronicity of inflammation in vivo. Further studies are required to elucidate critically important steps in TNF signaling that might be dysregulated. This will lead to a better understanding of the pathogenesis of these diseases and poteintially reveal new, more specific therapeutic targets.
Keywords: TNF-alpha, TNF-R2, signal transduction, inflammatory disorders.
Immunochemistry
- The cGMP Synthesis and PKG1 Expression in Murine Lymphoid Organs
Ewa Kurowska1, Marcin Kobiałka1, Ewa Zioło2, Leon Strządała2 and Wojciech A. Gorczyca1, 3 (1Laboratory of Signaling Proteins and 2Laboratory of Cellular Immunology, L. Hirszfeld Institute of Immunology and Experimental Therapy, Polish Academy of Sciences, Wrocław, Poland, 3Department of Biogenic Amines, Polish Academy of Sciences, Łódź, Poland)
Abstract. Numerous reports indicate that cyclic 3′,5′ guanosine monophosphate (cGMP) is involved in the regulation of immune processes. However, the mechanisms responsible for the synthesis of this nucleotide and its signaling pathways in immune cells are still not well recognized. The aim of our study was to establish: 1) which form of guanylyl cyclase synthesizes cGMP in murine lymphoid organs and 2) whether the same organs express the isoforms PKG1alpha and/or PKG1beta of protein kinase G, known as a possible target for synthesized cGMP. Cells isolated from thymus, lymph nodes, and spleen were treated with activators (SNP, ANP, CNP, STa) of soluble or particulate cyclases. Sodium nitroprusside (SNP) elevated intracellular cGMP 2-fold in thymic and lymph node cells and about 10-fold in spleen cells. Atrial natriuretic peptide (ANP) caused modest but statistically significant increases of cGMP in cells of all the organs. Additionally, spleen cells elevated their cGMP content about 2-fold in response to C-type natriuretic protein (CNP). In cellular homogenates of all the analyzed organs, the antibody anti-PKG1beta stained the 78 kDa band corresponding to the molecular mass of PKG1. Only homogenates of spleen cells were stained by the antibody recognizing PKG1alpha. Our results indicate that in all the investigated organs, cGMP may be synthesized mainly by soluble guanylyl cyclases in response to nitric oxide. The modest increase of cGMP upon stimulation by ANP suggests that in all these organs either exist only a small subpopulation of cells that express particulate cyclase GC-A or GC-A is expressed at very low level. In spleen cells, however, cyclase GC-B appears to be the more active enzyme. Elevated cGMP concentration may in turn activate PKG1beta in thymus, lymph node, and spleen cells and also PKG1alpha in spleen cells.
Keywords: cyclic nucleotides, guanylyl cyclases, protein kinases, signal transduction, lymphoid organs.
Vol. 50, No. 5, 2002
CONTENTS
Review
- Immunotherapy of Cancer through Targeting of the p53 Tumor Antigen
Sander Zwaveling1, 2, Sjoerd H. Van der Burg1, Anand G. Menon, Cornelis J. M. Melief1 and Rienk Offringa1 (1Leiden University Medical Center, Department of Immunohematology and Blood Transfusion, Tumor Immunology Group and 2Department of Surgery, Albinusdreef 2, 2333 ZA Leiden, The Netherlands)Abstract. The expression of the p53 tumor suppressor protein is frequently increased in a great variety of human cancers, making this antigen an attractive candidate for targeting therapeutic T cell immunity. However, potential complications as a result of immunological tolerance or auto-immune pathology must be taken into account when exploiting this ubiquitously expressed auto-antigen for immunotherapy of cancer.
Keywords: cancer immunotherapy; p53 tumor antigen.
- Lymphocytes Distribution and Intrahepatic Compartmentalization during HCV Infection: a Main Role of MHC-Unrestricted T Cells
Chiara Agrati, Carla Nisii, Alessandra Oliva, Gianpiero D’Offizi, Carla Montesano, Leopoldo Paolo Pucillo and Fabrizio Poccia (National Institute for Infectious Diseases „L. Spallanzani”, Rome, Italy)Abstract. Hepatitis-C virus (HCV) infection induces an acute and chronic liver inflammation through an immune mediated pathway that may lead to cirrhosis and liver failure. Indeed, HCV-related hepatitis is characterized by a dramatic lymphocyte infiltrate in the liver which is mainly composed by HCV non-specific cells. Several data indicated that IFN-gamma secretion by intrahepatic lymphocytes (IHL) may drive non specific cell homing to the liver inducing IP-10 production. An interesting hallmark of these IHL is the recruitment of lymphocytes associated with mechanism of innate immunity such as NK, NKT and gamma delta T lymphocytes. CD81 triggering on NK cell surface by the HCV envelope glycoprotein E2 was recently shown to inhibit NK cell function in the liver of HCV-infected persons resulting in a possible mechanism contributing to the lack of virus clearance and to the establishment of chronic infection. In contrast, intrahepatic NKT cells restricted to Cd1d molecules expressed on the hepatocyte surface may contribute to a large extent to the liver damage. Finally, an increased frequency of T cell expressing the gamma delta TCR was observed in HCV-infected liver and recent observations indicate that intrahepatic gamma delta T cell activation could be directly induced by the HCV/E2 particle through CD81 triggering. These cells are not HCV specific, are able to kill target cells including primary hepatocytes and their ability to produce Th1 cytokines is associated with an higher degree of liver disease. Altogether, CD1d/NKT and/or E2/CD81 interactions may play a major role in the establishment of HCV immunopathogenesis. In absence of virus clearance, the chemokine-driven recruitment of lymphocytes with an innate cytotoxic behavior in the liver of HCV infected patients may boost itself leading to the necroinflammatory and fibrotic liver disease.
Keywords: hepatitis C, liver, intrahepatic lymphocytes, NK cells, NKT cells, NT cells, gamma delta T lymphocytes, HLA, CD1, CD81, E2.
- Immunotherapy in the Management of Sepsis
Janusz Piotr Sikora (Department of Pediatric Propedeutics, Institute of Pediatrics, Medical University of Łódź, Sporna 36/50, 91-738 Łódź, Poland)Abstract. The work presents the role of Gram-negative bacteria endotoxins, pro- and anti-inflammatory cytokines and reactive oxygen species (ROS) in the complex and not fully explained pathogenesis of sepsis. The so called ‘respiratory burst’ of neutrophils and antioxidant mechanisms of the host are also discussed. The work has focused on possible approaches to the management of sepsis connected with immunotherapy. Neutralisation of endotoxin lypopolysaccharide (LPS), anti-TNF-alpha therapy with monoclonal antibodies or pentoxifylline (PTXF) as well as soluble recombinant cytokine agonists and antagonists used in clinical trials were taken into consideration. Besides, cytokine manipulation therapy, anti-adhesion techniques or glicocorticoides and antioxidant barrier interference were also described. So far there has been no immunotherapy of sepsis in children of proven clinical efficacy, which prompts aggressive examination of the immune system, aimed at affecting its function.
Keywords: sepsis, cytokines, reactive oxygen species, monoclonal antibodies, cytokine agonists and antagonists, immunotherapy.
- The Role of TGF-Beta Signaling in the Pathogenesis of Fibrosis in Scleroderma
Hironobu Ihn (Department of Dermatology, Faculty of Medicine, University of Tokyo, Tokyo, Japan)Abstract. Excessive extracellular matrix (ECM) deposition in the skin, lung, and other organs is a hallmark of systemic sclerosis (SSc). The pathogenesis of SSc is still poorly understood, but increasing evidence suggests that transforming growth factor (TGF)-b is a key mediator of tissue fibrosis as a consequence of ECM accumulation in pathologic states such as systemic sclerosis. TGF-b regulates diverse biological activities including cell growth, cell death or apoptosis, cell differentiation, and extracellular matrix (ECM) synthesis. TGF-b is known to induce the expression of ECM proteins in mesenchymal cells, and to stimulate the production of protease inhibitors that prevent enzymatic breakdown of the ECM. This review focuses on the possible role of TGF-b in the pathogenesis of fibrosis in SSc.
Keywords: TGF-beta; fibrosis; systemic sclerosis; signal transduction.
Clinical Immunology
- A Protective Role of HLA-DR Locus in Patients with Various Clinical Types of Alopecia Areata
Grażyna Broniarczyk-Dyła1, Magdalena Prusińska-Bratoś1, Małgorzata Dubla-Berner1, Celina Arkuszewska1, Maciej Borowiec2, Marek L. Kowalski2 and Grzegorz Woszczek2 (1Department and Clinic of Dermatology and Venereology, Medical University of Łódź, Krzemieniecka 5, Łódź, Poland, 2Department of Clinical Immunology and Allergy, Medical University of Łódź, Poland)Abstract. One of genetic factors associated with the development of alopecia areata (AA) is HLA locus. The study comprised 52 patients with AA, aged 10 to 64 years. The frequences of HLA-DR B alleles in patients and controls were compared. The control group comprised 152 healthy persons. Familiar occurrence of alopecia areata was seen in 7 cases (13.5%). Significantly lower frequency of HLA-DRB1*03 was observed in patients with AA in comparison to control group. In all patients with AA alleles HLA-DRB1*15/*16 occurred more frequently than in control group, but it was not significant after correction. In the group of patients with more severe form of alopecia areata (AAT/U) there was no significant difference in HLA- DR alleles distribution.
Keywords: alopecia areata, familiar inheritance, HLA locus.
Experimental Immunology
- Lack of Relation between Serum Content of MBL, sCD14, Anti-PPD and Anti-Hsp65 IgG, and Ingestion of Mycobacterium bovis BCG Bacilli by Phagocytes
Beata Paziak-Domańska1, Agnieszka Bonar1, Magdalena Kowalewicz-Kulbat1, Magdalena Klink2, Michał Kowalski3, Jari Karhukorpi4, Ritta Karttunen5, Magdalena Jurkiewicz1, Barbara Różalska1 and Wiesława Rudnicka1 (1Department of Immunology and Infectious Biology, University of Łódź, Banacha 12, 90-237 Łódź, Poland, 2Microbiology and Virology Centre, Polish Academy of Science, Łódź, Poland, 3Health Centre for High Schools, Łódź, Poland, 4Clinical Microbiology Laboratory, University Hospital of Oulu, Finland, 5Department of Medical Microbiology, University of Oulu, Finland)Abstract. Prophylactic vaccination against tuberculosis (TB) with a live attenuated strain of Mycobacterium bovis Bacille Calmett’e-Gúerin (BCG) has been used worldwide. However, TB remains one of the most significant diseases of humans and animals. Better understanding of the mechanisms of human immunity to mycobacteria is essential for development of new vaccines and estimation of their efficacy. In this study we determined the levels of known humoral mediators of mycobacterial phagocytosis – mannose binding lectin (MBL), soluble CD14 (sCD14), antibodies of IgG class against mycobacterial purified protein derivative (PPD) and mycobacterial Hsp65 antigen, in the sera from healthy young volunteers vaccinated with BCG and presenting positive and negative Mantoux responses to PPD. Than we asked a question as to whether macrophages and polymorphonuclear leukocytes (PMNs) from the individuals with positive (TT(+)) and negative (TT(-)) tuberculin tests differ by the ability to ingest mycobacteria. Also we were looking for a relation between the intensity of mycobacterial ingestion by phagocytes in the medium with autologous sera containing different concentration of MBL, sCD14 andf anti-mycobacterial IgG. We found no significant differences between the investigated parameters for TT(+) and TT(-) volunteers. Our result suggest that ability of macrophages and PMNs to ingest mycobacteria depends on an individual intrinsic capacity of phagocytes.
Keywords: tuberculin test, macrophages, polymorphonuclear leukocytes.
Immunochemistry
- Structural and Serological Characterisation of the Lipopolysaccharide from Proteus penneri 20 and Classification of Cross-Reacting Proteus penneri Strains 10, 16, 18, 20, 32 and 45 in Proteus Serogroup O17
Zygmunt Sidorczyk1, Filip V. Toukach2, Krystyna Zych1, Nikolay P. Arbatsky2, Dominika Drzewiecka1, Andrzej Ziółkowski1, Alexander S. Shashkov2 and Yuriy A. Knirel2 (1Department of General Microbiology, Institute of Microbiology and Immunology, University of Łódź, Banacha 12/16, 90-237 Łódz, Poland, 2N. D. Zelinsky Institute of Organic Chemistry, Russian Academy of Sciences, Leninsky Pr. 47, Moscow 119991, Russia)
Abstract. O-specific polysaccharide (O-antigen) of the lipopolysaccharide of Proteus penneri 20 was studied using sugar analysis along with various one- and two-dimensional NMR spectroscopy techniques. The following structure of the polysaccharide was established. It has the same carbohydrate backbone structure as that described earlier for P. penneri 16, in which the positions of the O-acetyl groups have not been determined. P. penneri 20 O-antiserum showed a strong cross-reactivity with the lipopolysaccharides of P. penneri 10, 16, 18, 32, 45 and P. mirabilis O17. These data enable classifying these strains together with P. penneri 20 in one Proteus serogroup, O17.
Keywords: lipopolysaccharide, O-antigen, bacterial polysaccharide, O-acetyl group, serological classification, O-serogroup, Proteus penneri.
Vol. 50, No. 6, 2002
CONTENTS
Review
- Stare i Nowe Sposoby Leczenia Chorób Zakaźnych: Wirusy Wszędzie i Nigdzie
Old and New Prescriptions for Infectious Diseases and the Newest Recipes for Biomedical Products in Plants (in Polish)
Hilary Koprowski, (Biotechnology Foundation Laboratories at Thomas Jefferson University, Philadelphia, Pennsylvania, USA)Abstract. The three antiviral vaccines discovered in the 18th century (smallpox), 19th century (rabies), and 20th century (polio) share a common feature: none would ever be licensed today for human vaccination. Yet Jenner’s smallpox vaccine led to the eradication of smallpox, Pasteur’s rabies vaccine represented the first successful post-exposure treatment of people bitten by rabid animals, and polio vaccine administered since its discovery in 1950 is leading to the eradication of polio (in the years 2004-2005) from the earth. However, in the case of rabies, efforts at complete eradication are unrealistic, despite the availability of a very effective vaccine, since rabies, unlike smallpox and polio, is not limited to humans and can infect all domestic and wild mammalian species. Rabies is probably the oldest known infectious disease, yet knowledge of the virus and the disease is far from complete. For instance, the appearance of 24 cases of „cryptic” rabies in the USA, i.e. cases not associated with any bite or scratch, with an incubation period in humans extending 6-8 years, is a puzzling phenomenon that cannot be readily explained. On the other hand, rabies is one of the few strictly neuronal infections and, as such, is an excellent model for the study of neurotropic virus distribution in the brain. Apoptosis induced by a rabies strain expressing high levels of glycoprotein spreads much more slowly through brain tissue than that induced by strains producing lower glycoprotein levels. Attenuated rabies virus constructed to express twice the normal glycoprotein levels is also an excellent antigen for induction of immune responses in the host. Foreign antigens using this vector may also produce highly immunogenic vaccines.
Global Approach to Immunization. Those monitoring the spread of AIDS in many parts of the world know that cost of treatment is one of the major problems in combating the disease. Vaccines against HIV face the same problem. In general, the price of vaccines and sera is exorbitant for the afflicted population in developing countries. In addition, the dearth of syringes, the unavailability of nurses and doctors to administer multiple vaccine injections, and other factors in these countries require a drastic change in current vaccine production approaches. About 12 years ago, plants became vehicles to produce biomedical reagents. Plants can be exposed directly to a construct containing a foreign gene and Agrobacterium to create a transgenic plant that, over several generations, produces the desired product. Alternatively, plants infected with a plant virus (e.g. alfalfa mosaic virus) fused with a foreign gene can propagate the foreign antigen as the virus multiplies. Extraction of the plant virus followed by purification provides the desired biomedical product. Our use of either of these systems has led to the creation of plants producing vaccines, sera, hormones, and other biological reagents. In two clinical trials at the Institute of Bioorganic Chemistry of the Polish Academy of Sciences in Poznan, volunteers who ingested lettuce expressing hepatitis B vaccine showed hepatitis B antibodies in their sera. In another trial carried out at the Biotechnology Foundation Laboratories in Philadelphia, volunteers ingesting a spinach-rabies vaccine showed an immunological priming effect, since only one injection of commercially available rabies vaccine significantly raised the level of rabies-specific antibodies. Vaccines against HIV gp120 and Tat have been produced in spinach, and a construct of gp120 with the CD4 receptor is now being adapted to this plant. Two types of antibodies against rabies and against colorectal cancer are being produced in tobacco and in lettuce. The suboptimal quality of the currently available anthrax vaccine prompted our efforts to produce the anthrax Protective Antigen (PA)in tobacco and lettuce. Quite clearly, plants will play a prominent role in producing a variety of biomedical reagents in the future.Keywords: vaccines; viruses; infectious diseases; bioproducts in plants.
- Predicting Outcome in Haematological Stem Cell Transplantation
Anne M. Dickinson1, James Cavet2, Hannah Cullup1, Xiao N. Wang1, Mark Jarvis1, Libset Sviland2 and Peter G. Middleton3 (1University Departmentt Haematology, Tyneside Leukaemia Research Laboratory, Royal Victoria Infirmary, Queen Victoria Road, Newcastle upon Tyne, UK, 2Avdeling for Patologi, Haukeland Sykehus, Bergen, Norway, 3Leukaemia Research Fund Laboratories, The Medical School, University of Newcastle, Newcastle-upon-Tyne, UK)Abstract. This present review summarises recent results investigating the role of certain cytokine gene polymorphisms, including TNFa, IFNg, IL-6, IL-10 and IL1 Ra, in allogeneic stem cell transplantation. The review discusses their role in predicting outcome and the development of a genetic risk index for Graft versus Host Disease in HLA matched sibling transplants. By the comparative use of an in vitro human skin explant model initial results suggest that certain cytokine gene polymorphisms may be associated with more severe disease.
Keywords: cytokine gene polymorphisms; predicting GvHD; an in vitro human skin explant model.
- Structures and Serology of the O-Specific Polysaccharides of Bacteria of the Genus Citrobacter
Yuriy A. Knirel1, Nina A. Kocharova1, Olga V. Bystrova1, Ewa Katzenellenbogen2 and Andrzej Gamian2 (1N.D. Zelinsky Institute of Organic Chemistry, Russian Academy of Sciences, Leninsky Prospekt 47, Moscow, 119991, Russia, 2Institute of Immunology and Experimental Therapy, Polish Academy of Sciences, Weigla 12, 53-114 Wrocław, Poland)Abstract. The review presents the structures of the O-specific polysaccharides (O-antigens) of the lipopolysaccharides isolated from over 25 Citrobacter strains, which represent different species and serogroups. The correlation between O-antigen structure and immunospecificity as well as numerous cross-reactions between Citrobacter and other enterobacterial species are discussed.
Keywords: Citrobacter; O-antigen; lipopolysaccharide; structure; immunospecificity
- Streptomyces and Escherichia coli, Model Organisms for the Analysis of the Initiation of Bacterial Chromosome Replication
Walter Messer1 and Jolanta Zakrzewska-Czerwińska2 (1Max-Planck-Institut für molekulare Genetik, D-14195 Berlin, Germany, 2Institute of Immunology and Experimental Therapy, Polish Academy of Sciences, Wrocław, Poland)Abstract. Streptomyces coelicolor A3(2) and Escherichia coli are quite different in their life-style and the structures of their genomes. Streptomyces exhibit complex multicellular development including formation of multigenomic hyphae during growth. These organisms possess a large linear (8.7 Mb) and GC-rich (~72%) chromosome. The genome sequence of S. coelicolor has just been completed. The difference between E. coli and Streptomyces making them an excellent model organisms for a comparison of their replication modes. In this review, we compare initiation of chromosome replication in both organisms. Their replication origins are different in size, but both have DnaA boxes – a binding motifs for initiator DnaA protein. The two DnaA proteins have practically the same biochemical properties. Many aspects of the control of initiation seem to be similar. A comparison of the two systems thus allows us to define those aspects of replication initiation that are universally used in the eubacterial kingdom.
Keywords: DnaA; Escherichia coli; oriC; Streptomyces coelicolor.
- Lactoferrin and Immunologic Dissonance: Clinical Implications
Marian L. Kruzel1 and Michał Zimecki2 (1Department of Integrative Biology and Pharmacology, University of Texas Health Science Center, Houston, TX 77030, USA, 2Ludwik Hirszfeld Institute of Immunology and Experimental Therapy, Polish Academy of Sciences, Wrocław, Poland)Abstract. Homeostasis is the maintenance of equilibrium in a biological system by means of positive and negative feedback control mechanisms that counteract influences tending toward physiological dissonance. At the molecular level, homeostasis is controlled by the network of the neuro-endocrine-immune system, in which lactoferrin plays a central role. The purpose of this review is to provide a comprehensive summary of a collaborative study established between the Hirszfeld Institute of Immunology and Experimental Therapy (Wrocław, Poland) and the University of Texas Health Science Center (Huston, USA) regarding lactoferrin and its role in homeostasis. In our studies we focused on the immunoregulatory functions of lactoferrin, both in vitro and in vivo,. We investigated the immune status of individuals subjected to different insults, including experimental endotoxemia in mice and surgery in humans. We also studied a lactoferrin-dependent delayed type hypersensitivity (DTH) response to evaluate some of the mechanisms by which lactoferrin can effectively substitute an adjuvant in vaccine.
Keywords: lactoferrin; inflammation; immunomodulation; oxidative stress; anti-aging.
- Experimental Therapies for Psoriasis
Khusru Asadullah1, Hans-Dieter Volk2, Markus Friedrich3 and Wolfram Sterry3 (1Corporate Research Business Area Dermatology, Schering AG, D-13342 Berlin, Germany, 2Department of Immunology, 3Department of Dermatology and Allergology, University Hospital Charité, Berlin Humboldt University, D-10098 Berlin, Germany)Abstract. There is a high medical need for better therapies for psoriasis. Based on new insight into the pathophysiology of this frequent immune disease, a number of novel systemic immunomodulatory therapies are currently in clinical development. These include approaches targeting antigen presentation and costimulation, T cell activation and leukocyte adhesion, action of proinflammatory mediators, and modulating the cytokine balance. Although mainly only preliminary data are available so far, these trials contribute to a further understanding of the disease and will eventually lead to new therapeutic options for psoriasis. Moreover, since psoriasis can be considered as a visible model disease for T cell-mediated disorders characterized by a type 1 cytokine pattern in general, such approaches may have impact for other immune disorders as well. Here we review the rationale and the initial clinical data of these important recent experimental therapies.
Keywords: psoriasis; immunotherapy; cytokines; costimulation; biologicals.
Original Paper
- Diminished Expression of the Type II Receptor for TGFb (TGFbRII) in T Lymphocytes from Patients with Sezary Syndrome Is Not Due to Mutations in the Receptor’s Poly-A Tract: Limitations of the Standard RT-PCR in cDNA Sequence Analysis of Homopolymeric Base Stretches
Qian Zhang1, Renold J. Capocasale1, Floyd E. Fox2, Vahe Bedian3, Eric C. Vonderheid4, Alain Rook2, Jonni S. Moore1, Peter C. Nowell1, Dale S. Haines5 and Mariusz A. Wasik1* (1Department of Pathology and Laboratory Medicine, 2Dermatology, and 3Department of Genetics, University of Pennsylvania Medical Center, Philadelphia, PA19104, USA, 4Department of Dermatology and 5Division of Hematology/Oncology, Hahnemann University)
Abstract. Peripheral blood lymphocytes from patients with Sezary syndrome (SzS) frequently demonstrate decreased surface expression of transforming growth factor b receptor II (TGFbRII). The mechanism of this low TGFbRII expression remains unknown. Because mutations within the poly-A tract of the TGFbRII sequence (nucleotides 709-718) were shown to result in diminished TGFbRII expression in other types of malignant tumors, we examined the sequence of the TGFbRII poly-A tract in two SzS-derived cell lines and in peripheral blood SzS cells from 17 SzS patients and 4 control, healthy individuals using DNA sequencing and single-stranded conformation polymorphism (SSCP) analysis. A standard bidirectional, automated sequence analysis of the RT-PCR-generated cDNA TGFbRII fragment showed a heterogenous population of the normal length, 10-, with admixed, shortened, 9-base poly-A stretches. Surprisingly, this mixture was present not only in the cells from 5 SzS patients and 2 SzS cell lines, but also in cells from 2 healthy control individuals. Importantly, the proportion of the shortened, 9-base fragments was markedly reduced or practically eliminated when the procedure was modified by usage of high-fidelity DNA polymerase, labeled primers and/or cloned RT-PCR products, which indicates that the presence of the shortened, 9-base fragments represented a procedural phenomenon rather than a true deletional mutation within an allele of the TGFbRII gene. Accordingly, SSCP analysis of genomic DNA did not reveal any mutations within the poly-A tract-containing region. These results indicate that a mechanism different from mutations in the polyadenine tract underlies the diminished TGFbRII expression in SzS cells and that the results of an unmodified, direct sequence analysis of homopolymeric base streaches in RT-PCR-derived cDNA should be interpreted with caution.
Keywords: type II receptor for TGFb;cutaneous T cell lymphoma; Sezary syndrome; poly-A tract sequence analysis.