CONTENTS
Review
- Classification of Cytokines According to the Receptor Code.
ANNA D.INGLOTAbstract. A simple classification of cytokines is presented. It is based on voluminous information available from published papers of many outstanding researchers of the cytokines and their receptors and/or summarized by experts in this field. The cytokďnes are divided into 7 families according to their receptor code. Such arrangement of apparently different cytokines may explain the biological significance of pleiotropy and redundancy. Furthermore, it may stimulate the development of the comprehensive classification of cytokines together with classical hormones, neuromediators, neuromodulators and related substances that participate in complex biological communication system.
Keywords: cytokines; hormones; receptors; signaling molecules; classification; nomenclature.lature.
- Immunomodulatory Role of Opioids in the Central Nervous System.
WEN S. SHENG, SHUXIAN HU, GENYA GEKKER, SHENG ZHU, PHILLIP K. PETERSON and CHUN C. CHAOAbstract. . Endogenous opioid peptides play a variety of roles in the central nervous system (CNS) from development to immune modulation. These functions are mediated mostly via specific opioid receptors uni- quely localized in different brain regions and cells. Exogenous opioids can influence and modulate neuronal and glial cell function via an opioid receptor mediated mechanism, leading to either protection or damage of the brain. Mechanisms underlying CNS opioid effects may be mediated via immune mediators, such as cytokines, ß-chemokines, and free radicals (i.e. reactive oxygen intermediates (ROI) .and nitric oxide (NO)) produced by activated glial cells (microglia and astrocytes). In the pathogenesis of HIV-1 infection in drug addicts, opiates such as morphine have been postulated to promote the progression of this virus and the development of secondary opportunistic infections. Kappa opioid receptor (KOR) ligands, on the other hand, may play a neuroprotective role. Diferences in species, age, sex, cell culture system, stimuli, opioid admini- stration route, concentrations used, strain of infectious agents, and treatment regimes have contributed to many conflicting results in the field of opioid research. More studies are needed to delineate how opioids exert their effects on glial cells as well as neurons with the goal of finding new therapeutic approaches for neuro- degenerative diseases, such as AIDS dementia.
Keywords: opioids; immunomodulation; central nervous system; neurotoxicity; HIV.y; HIV.
- Gene Therapy: Panacea or Placebo? I. Strategies and Limitations of Gene Therapy.
DARIUSZ C. GÓRECKI and KAY D. MACDERMOTAbstract. .Progress in genetics and molecular medicine has led to characterization of many disorders at the level of specific genes. Techniques for reliable diagnosis of these disorders have been developed in parallel. Attempts are currently being made to develop DNA-based therapeutic procedures to correct genetic diseases. These procedures are known under a common name of gene therapy. The initial step in gene therapy is the delivery of the gene of interest into target cells. There are several conceptually different approaches to achieve this, e.g. targeting can be accomplished by using viral vectors (transduction) or DNA-mediated routes (trans- formation) either ex vivo or in vivo. The selection of vector systems and the choice of delivering genes either into isolated cells or directly in the organism depend on factors like: the nature of target cell or organ, the levels of expression required, stability and/or regulation of expression, safety, etc. Many viruses have been adapted for use as vectors for gene therapy, according to their specific properties. Viral genomes have been modified to remove their ability to replicate and to increase the cloning capacity. Non-viral gene delivery systems rely on cellular mechanisms to import DNA into the cell nucleus and different methods to enhance DNA uptake have been attempted. Despire many significant achievements there are still obstacles to the development of effective clinical products. Most significant are the low levels and stability of expression of introduced genes and immune responses to vectors and/or gene products.
Keywords: gene therapy; viral vectors; gene targeting; direct DNA injection; liposomes; immunogenicity.nicity.
- Gene Therapy: Panacea or Placebo? II. Main Applications of Gene Therapy.
DARIUSZ C. GÓRECKI and KAY D. MACDERMOTAbstract. . For most disorders the ideal goal of gene therapy is the repair of the mutated gene in the target tissue. However, the techniques required for such an approach are still at an early stage of development. Most current research is directed towards delivery of normal gene sequences in order to generate active protein and compensate for the lack of endogenous production. This approach may be suitable for the treatment of recessive monogenic disorders, but is inappropriate for dominantly inherited disorders. Therefore, gene thera- py was originally intended as a most promissing approach for the treatment of inborn errors of metabolism. However, apart from the correction of heritable genetic disorders, gene delivery has many potential ap- plications including treatment of malignancies, atherosclerosis and vascular proliferative disorders, rheumato- id arthritis and viral infections. The authors discuss significant examples marking progress in the development of gene therapy for specific diseases, present some hopes and hurdles that have arisen from some recent preclinical and early clinical trials.
Keywords: gene therapy; muscular dystrophy; cystic fibrosis; immunodeficiences; neurodegeneration; lysoso- mal storage disorders.orders.
- Sirolimus in Transplantation.
STANISŁAW M. STEPKOWSKI, LING TIAN, MOU-ER WANG, XIUMEI QU, KIMBERLY NAPOLI and BARRY D. KAHANAbstract. . Although sirolimus (SRL) alone was very effective in rodents and pigs, it produced toxic side effects in dogs. Low doses of SRL combined with cyclosporine/brequinar (CsA/BQR) combinations achieved potent immunosuppression in the CsA-resistant mouse model. Similarly, SRL/CsA/BQR therapy protected kidney allografts from rejection in dogs without producing toxic side effects. In the CsA-sensitive rat model SRL/CsA combinations produced a potent synergistic interaction. In addition, recent clinical trials document the benefi- cial effects of low SRL doses in human kidney transplant recipients. Sirolimus, when combined with standard immunosuppressive therapy, remarkably reduces the incidence of acute rejection and permits individual drug dose reduction.
Keywords: cyclosporine; graft-versus-host; sirolimus; tacrolimus.s; tacrolimus.
- Diphtheria Toxin as a Molecular Tool in the Study of Acidic Fibroblast Growth Factor Slgnalllng.
ANTONI WIĘDŁOCHAAbstract. In eukaryotic cells proteins are translocated across a number of cellular membranes into various intracellular organelles such as the endoplasmatic reticulum, mitochondria, peroxisomes and chloroplasts. In all these cases the proteins are translocated away from the cytosol. However, certain proteins are also translocated in the opposite direction, from the exterior to the cytosol. Well established examples are some bacterial and plant protein toxins, that exert their effect in the cytosol. A common property of protein toxins with intracellular action is that they contain two functionally different moieties, in many cases consisting of two disulfide-linked polypeptides. Relatively little is known about how these proteins cross the membrane. The translocation process is best understood in the case of diphtheria toxin, which binds to cell surface receptors, is then taken up by endocytosis and is subsequently translocated to the cytosol, where it inactivates elongation factor 2. Recently it has been recognized that diphtheria toxin as well as a few other protein toxins can be used to carry passenger peptides or proteins into cells (in addition to other usefull roles which the toxins have begun to play in understanding many cellular processes and in certain prophylactic and therapeutic purposes). Here, the approach of using diphtheria toxin as a translocation vehicle in the study of new aspects of signal transduction mechanisms activated by acidic fibroblast growth factor is discussed and the possibility that some proteins have distinct functions in more than one cellular compartment is considered. Finally, this article focuses on the role of the toxins as tools in cell biology and experimental medicine.
Keywords: protein toxins; diphtheria toxin; membrane translocation; acidic fibroblast growth factor signall- ing; nuclear targeting.ear targeting.
- The Use of Tissue Slices in Immunological In vestigations.
GRZEGORZ SKIBIŃSKI and KEITH JAMESAbstract. Our knowledge of cellular and molecular events taking place during various immune responses comes mainly from in vitro studies where isolated cells are exposed to defined stimuli. This reductionist approach has greatly advanced our understanding of the immune system. However, these studies do not truly reflect the complexity of the integrative events which take place in both primary and secondary lymphoid tissue in vivo. In order to address this problem we have developed a tissue culture procedure which involves the cultivating of precision cut human spleen slices at gas/liquid interface. We have shown, that cells in this culture system show marked differences in cytokine and immunoglobulin production in comparison with conventional single cell suspension cultures, obtained from the same spleen and run in parallel. In this review article we describe the basic technique, results which we have obtained in this system and discuss the possible basis of the observed differences.
Keywords: tissue slices; in vitro culture; lymphoid organs; research applications. applications.
Immunochemistry
- Immunomodulatory Activity of G-Actin Fragments Containing a Thymopentin-Like Sequence.
IGNACY Z. SIEMION, IWONA STRUG, ALEKSANDRA WYSŁOUCH-CIESZYŃSKA, EWA NOWAK and ZBIGNIEW WIECZOREKAbstract. A discontinuous thymopoietin-like motif, composed of the fragments 97 -111 and 277 – 307 of the molecule, as well as of residues Arg 178 and Asn 163 of G-actin was found. It was established that G-actin has an immunosuppres’sive activity regarding the humoral immune response. This activity is probably connected to the thymopentin-like sequence RKDLY, which is present in the 277 – 307 fragment of G-actin. The im- munomodulatory activity of a series of peptide-partial sequences of G-actin was tested using plaque-forming cells (PFC) and delayed type hypersensitivity (DTH) tests. The investigated series consisted of five peptides: RKDLY (I), RKDLYANT (II), DVDIRKDLY (III), DVDIR (NOz)KDLY (IV), DVDIRKDLYANT (V). The peptides have the immunosuppressive activity regarding the humoral and cellular immune response.
Keywords: G-actin; thymopoietin; thymopoietin-like motif. motif.
- Immunomodulatory Activity of Oligopeptides Related to Interleukin 1 Receptor Antagonist Sequence.
ALICJA KLUCZYK, IGNACY Z. SIEMION, JACEK SŁOŃ-USAKIEWICZ and ZBIGNIEW WIECZOREKAbstract. We examined the immunomodulatory properties of peptides from interleukin 1 receptor antagonist (IL-1Ra) with regard to the humoral (plaqueforming cells – PFC) and cellular (delayed type hypersensitivity – DTH) immune response and GvH reaction. It was found that peptide RKSSK (II) from the N-terminal part of IL-l Ra, although inactive with regard to the inhibition of IL-1-IL-1 receptor interaction, reduces immune response in a manner similar to cyclosporin A (DTH, PFC in vivo). Peptide GRKSSK (III) was even more potent, whereas peptides from respective fragment of mouse IL-1Ra were weaker immunosuppressants than II. Peptide VTKFYF (VII) from the C-terminal part of IL-lRa, very active as IL-1 inhibitor, and its analog VIII with Asp residue, characteristic for IL-l, instead of Lys from IL-lRa, showed only limited activity despite the previously observed competition with IL-1 for the cellular receptor. Thus, no correlation between the in- hibitory and immunomodulatory properties of peptides derived from IL-1Ra was observed.
Keywords: interleukin 1 receptor antagonist; immune response; tuftsin; thymopentin.pentin.
- Structural and Immunochemical Studies on the O-Specific Polysaccharide of Proteus penner Strain 15.
KRYSTYNA ZYCH, MARCIN KOWALCZYK, FILIP V. TOUKACH, NIKOLAY A. PARAMONOV, ALEXANDER S. SHASHKOV, YURIY A. KNIREL and ZYGMUNT SIDORCZYKAbstract. On the basis of sugar analysis and 1H- and 13C-NMR spectroscopy, it was shown that the O-specific polysaccharide of Proteus penneri strain 15 has a trisaccharide repeating unit, including an acetal-linked pyruvic acid residue, and is structurally identical to the capsular polysaccharide of Proteus vulgaris strain ATCC 49990. Serological studies supported this conclusion and demonstrated the presence in the homological antiserum of both anti-core and anti-O chain antibodies reacting with a lipopolysaccharide (LPS) epitope containing N-acetylglucosamine and galactose residues.
Keywords: Proteus penneri; lipopolysaccharide; O-specific polysaccharide; pyruvic acid acetal.acetal.
Experimental Oncology
- Detection of Soluble IL-2 Receptor in the Serum and Bone Marrow of Patients with Minimal Residual Hematological Malignaces: Induction under Therapy with IL-2.
MARTYNA KANDEFER-SZERSZEŃ, WOJCIECH LEGIEC, ANNA DMOSZYŃSKA and WOJCIECH RZESKIAbstract. We measured the soluble IL-2 receptor a (sIL-2Ra) in sera and in bone marrow of 20 patients with minimal residual hematological malignances, 6 of them with multiple myeloma (MM), 8 with Hodgkin’s disease (HD) and 6 with non-Hodgkin’s lymphoma (NHL), low grade. Compared to 10 normal individuals, HD and NHL group of patients had in sera significantly increased levels of sIL-2Ra (252.8±42.9 versus 14372±1639 and 761.8±431 U/ml, respectively). After low-dose IL-2 given subcutaneously once daily at a dose of 1.8 x 106 U/patient for 3 weeks, additional significant increase in levels of IL-2Ra was observed in sera and in bone marrow of patients with NHL (761.8 ± 431 versus 2633 ± 788 U/ml and 785 ± 448 versus 2475±431 U/ml, respectively). The increase of sIL-2Ra level after IL-2 therapy was also seen in sera and in bone marrow of HD and MM group; however, because of high standard deviation this increase was not statistically significant. We conclude that 1) in comparison to healthy subjects the levels of sIL-2Ra remained elevated in HD and NHL patients, even at the stage of minimal residual disease (MRD) after intensive chemotherapy or radiotherapy, 2) the levels of sIL-2Ra which appeared in sera and bone marrow of patients after IL-2 therapy seemed to be dependent on the type of hematological disorders.
Keywords: IL-2a receptors; IL-2 low-dose therapy.herapy.
- Release of Soluble Tumor Necrosis Factor Receptors from Polymorphonuclear Cells of Breast Cancer Patients (Short Communication).
EWA JABŁOŃSKA and ZOFIA PIETRUSKAAbstract. The release of soluble tumor necrosis factor receptors (sTNF-Rs): soluble tumor necrosis factor receptor I (sTNF-RI) and soluble tumor necrosis factor receptor II (sTNF-RII) by polymorphonuclear cells (PMNs) derived from patients with breast cancer were measured. Serum levels of sTNF-RI and sTNF-RII in patients group were also determined. Significantly higher values of sTNF-RI and sTNF-RII released by PMNs of patients were observed than those in the healthy subjects. There was no significant differences in the serum levels of sTNF-RI and sTNF-RII between breast cancer patients and healthy control. Presented study demostrate that the tumor bearing state, involving breast cancer, may induce an altered soluble cytokine receptors release from the peripheral blood PMNs characterized by increased concentrations of sTNF-RI and sTNF-RII in the culture supernatants of these cells in vitro.
Keywords: polymorphonuclear cells; soluble tumor necrosis factor; breast cancer.breast cancer.
Various
- Hepatitis C Virus (HCV) Serotype in the Asymptomatic HCV-Infected Patients from Selected Groups (Short Communication).
MAŁGORZATA PAWŁOWSKA, WALDEMAR HALOTA, FRANCISZEK BULIK and EWA TOPCZEWSKA-STAUBACHAbstract. . In asymptomatic hepatitis C virus (HCV)-infected blood donors and persons from high risk groups (intravenous drug users and hemodialyzed patiénts) HCV serotypes 1, 2, 3, 4 and 6 have been detected. The most frequent was serotype l. In the group of intravenous drug users (IDU) a coexistence of different serotypes was observed.
Keywords: hepatitis C virus (HCV)-serotypes; asymptomatic HCV-infection; blood donors; intravenous drug users; hemodialyzed patients.yzed patients.
- Influence of Recipient Pretreatmerlt with Do nor Spleen Cells and 15-Deoxyspergualin on Rat Skin Graft Survival
MACIEJ KRZYMAŃSKI, ANDRZEJ OKO, ANA MARIA WAAGA, ELŻBIETA PAWLICZAK and KRZYSZTOF WIKTOROWICZAbstract. Experimental studies performed in rat kidney transplantation showed that treatment with 15-deoxy- spergualin (15-DOS) for 14 days after grafting may induce both permanent graft function and specific immune tolerance. The aim of this study was to check if donor spleen cell transfer and 15-DOS pretreatment before transplantation prolong skin graft survival. Pretreatment of the recipient with donor cells induced slight prolongation of subsequent skin graft survival. Addition of 15-DOS pretreatment for 14 days in a dose of 2.0 mg/kg b.w. to cell transfer did not inlluence survival of the graft. On the other hand, in the recipients receiving allogeneic spleen cells before transplantation and subtherapeutic doses of 15-DOS directly after grafting thé prolongation of skin graft survival was observed. These results indicate that pretreatment of the recipients with donor cells before allogeneic skin transplantation allows for reduction of immunosuppression after grafting.
Keywords: 15-deoxyspergualin; donor specific transfusion.c transfusion.
- Parvovirus Particles in a Fetal-Heart with Myocarditis: Ultrastructural and Immunohistochemical Study (Short Communication).
MARIA RESPONDEK, JOLANTA BRATOSIEWICZ, TOMASZ PERTYŃSKI and PAWEŁ P. LIBERSKIAbstract. We report on the occurrence of parvovirus particles and VP1 (84 kDa) and VP2 (58 kDa) viral antigens in the heart of a case of fatal myocarditis in a fetus of a 26 year old women. Numerous cells containing intranuclear inclusions were identified within the blood vessels of the heart in a close apposition to muscle fibers. These cells were characterized by plentiful mitochondria and were consistent with erythroblasts. Typically, inclusions consisted of electrondense marginated chromatin and granular and amorphous „cores”. At higher magnification, parvovirus particles, approximately 23 nm in diameter, were visualized either as relatively small clusters or forming large paracrystalline arrays. Virus buds were never observed. In addition, unusual membrane proliferation was seen. These findings support a notion that parvovirus may invade the fetal heart.
Keywords: parvovirus; B19; fetal myocarditis.l myocarditis.
- Natural and Probe Fluorescence of Lymphocytes of Patients with Pulmonary Tuberculosis Exposed to Radiation as a Result of the Chernobyl Nuclear Accident (Short Communication).
GENNADY L. GUREVICH, ALEXANDER V. VOROBEY, ELENA S. LOBANOK and NATALIA A. SHUKANOVAAbstract. Parameters of natural and probe fluorescence of peripheral blood lymphocytes have been studied in 25 normal persons and 72 patients suffering from extensive forms of pulmonary tuberculosis who live in various radioecological conditions. It was found that in cell suspension the intensity of fluorescence of protein and the membrane-bound merocyanine 540 dye was 2.0 – 3.2 and 3.9 – 7.1 times higher, respectively, for the lymphocytes of the examined patients. For lymphocytes of the patients who live in radionuclide-contaminated areas fluorescence of reduced pyridine nucleotides and some fluorescent probes (ANS, DPGT, the ratio of eximer to monomer fluorescence of pyrene) and the merocyanine-photosensitized cell death were also found to increase. These can indicate some changes of cell membranes and reduction of the oxidation resistance of lymphocytes. There was not any substantial dependence on the changes in the parameters determined in the patients calculated radiation doses was revealed. It was found that biophysical tests, in particular, photosen- sitized cell death, are informative for estimation of severity of the tuberculous process and prognosis of its outcome in patients who live in adverse radioecological conditions.
Keywords: tuberculosis; radiation; lymphocytes; lluorescent probes.probes.
- Interferon a Reduces the Level of Radiation-Induced Micronuclei in Mouse Bone Marrow Cells.
LUDMILA FOMENKO, NICOLAI SIROTA, VICTOR RAVIN and AZHUB GAZIEV
Abstract. The effect of natural mouse interferon a (IFN-a) on production of micronuclei (MN) in bone marrow cells of whole-body y-irradiated mice (in sublethal dose of 0.5 Gy from 6oCo source) was studied. It was shown that free interferon a (IFN-a(F)) and IFN-a encapsulated into liposomes (IFN-a(L)), when injected intraperito- neally to mice prior to their irradiation, reduced substantially the frequency of y-ray-induced micronuclei in polychromatic erythrocytes (MN-PCE) of the bone marrow of these mice. The antimutagenic effect of IFN-a(F), as registered by the micronucleus test, was observed in bone marrow cells of mice irradiated 24 h after injection. For IFN-a(L), the decrease in MN frequency in PCE of mice was observed 48-72 h after administration of the preparation.
Keywords: interferon a in liposomes; antimutagenicity; micronuclei; y-irradiated mice; in vivo.n vivo.