Vol. 45, No. 1, 1997

Reviews

Stem Cell Transplantation: Past, Present and Future.
E. DONNALL THOMAS

Abstract. The early attempts at human allogeneic marrow transplants in the 1950’s and 1960’s were largely unsuccessful. The probability of success has improved steadily in the past two decades. Cure rates now range from 90% for non-malignant diseases transplanted early to 15% for patients with advanced leukemia. Most marrow transplants have involved an HLA matched sibling donor but, more recently, a matched unrelated volunteer marrow donor can be found for many patients without a family donor. Current research is focused on new preparative regimens for elimination of malignant cells, better prevention of graft-versus-host disease, and the use of hematopoietic growth factors and cytokines. Autologous transplants, which use the patient s own marrow, are increasing, particularly for breast cancer. The hematopoietic stem cells are responsible for marrow regeneration after a transplant. Suff cient numbers of stem cells for transplantation can now be obtained from the peripheral blood after mobilization of these cells by chemotherapy or hematopoietic growth factors. Transplants can also be achieved using stem cells obtained from cord blood at the time of delivery, tissue typed, and cryopreserved for later use. A variety of technological advances has reduced the hospitaliza- tion time for transplant patients with a corresponding saving in cost.

Keywords: bone marrow transplantation; stem cells; cord blood. blood.
Role of Extracellular Matrix Proteins in Organ Transplantation.
JERZY W. KUPIEC-WEGLINSKI and ANDRZEJ GÓRSKI

Abstract. Recent data indicate that the extracellular matrix (ECM) proteins can regulate the process of T cell activation.Lymphocytes express an array of surface integrin and non-integrin receptors (adhesion molecules) mediating those phenomena.Since ECM proteins accumulate in situ during allograft rejection,it is likely that such T cell :ECM interactions are relevant in the immunopathology of rejection.Adhesion molecules are also thought to affect the very early events between host leukocytes and vascular endothelium.Therefore,immuno- modulation of T cell interactions with the ECM proteins and endothelium may lead to the development of nov- el therapeutic strategies in clinical organ transplantation.

Keywords: organ transplantation; extracellular matrix; adhesion molecules; vascular endothelium.helium.
Elastin Receptor and Cell-Matrix Interactions in Heart Transplant-Associated Arteriosclerosis.
ALEKSANDER HINEK

Abstract. Vascular cells, as well as monocytes, neutrophils, and lymphocytes which may infiltrate vascular walls and tissues express a multifunctional 67 kD protein which also serves as a subunit of the cell surface „elastin receptor”. This protein differs structurally and functionally from other matrix adhesion molecules. Unlike the integrins or cadherins, it is not a transmembrane molecule, but can be immobilized on the cell surface by association with two other membrane-anchored proteins. Once expressed on the cell surface, it may mediate cell-matrix interaction in a calcium-independent manner. Unlike most integrins, which recognize the linear sequence on the matrix ligands (RGD), it recognizes the secondary structure of the matrix macromole- cules and binds to several non identical domains on ditferent matrix components, as long as they form the appropriate hydrophobic conformation. Similarly to the transmembrane selectins, the 67 kD protein has lectin-like properties with the galactosugars’ binding specificity. However, binding of galactosugar-bearing ligands interrupts its contacts with matrix proteins and displaces the 67 kD protein from the cell surface. Moreover, the 67 kD protein also serves as an intracellular chaperone which facilitates secretion of tropoelas- tin and asserrbly of elastic fibers. In this review I will address the role of this 67 kD protein in mechanisms of mutual interaction between vascular smooth muscle cells, infiltrating leukocytes, and several components of extracellular matrix during the development of heart-transplant associated arteriosclerosis.

Keywords: arteriosclerosis; growth factors; elastin; extracellular matrix; fibronectin; interleukins; MHC.s; MHC.
Leukocyte Adhesion Defects.
AMOS ETZIONI

Abstract. Studies of genetic deficiency syndromes have provided important insights into the molecular basis and biology of leukocyte emigration. Both animal and human deficiency syndromes have been described.

Keywords: genetic deficiency syndromes; leukocyte adhesion deficiency; adhesion molecules;animal models ; gene therapy.herapy.

Cellular Immunology

Modified Cytotoxic T Lymphocyte Precursor Frequency Assay by Measuring Released Europium in a Time Resolved Fluorometer.
KHAWAJA HAQUE 1, CAROL TRUMAN 1, IAN DITTMER 2, GODFREY LAUNDY 1, PATRICIA DENNING-KENDALL 1,JILL HOWS 1, TERRY FEEST 2 and BENJAMIN BRADLEY 1

Abstract. The frequency of cytotoxic T lymphocyte precursors (CTLpf) can be quantified by using the principle of limiting dilution analysis (LDA). Chromium 51 (SlCr) and europium (Eu) release assays are based on the measurement of marker release after lysis of targets by the effector cells. Although, slCr release has been widely used to quantify cell lysis since its introduction, it has several disadvantages such as handling and disposal of radioisotopes as well as health risk to personnel involved performing the assay. This situation has led us to adopt a non-radioactive cytotoxicity assay. After 7 days culture the PHA-stimulated targets are labeled with europium DTPA chelate. Lysis of labeled targets by effectors releases the Eu-DTPA complex in culture medium – a highly fluorescent substance. The amount of fluorescence can be measured in a time resolved fluorometer. We describe here some modifcations of the original protocol which include optimising IL-2 requirements, reduction of incubation times, addition of an extra spin before 37oC incubation, readjust- ment of target cells per volume of labeling buffer and other crucial parameters increasing the specificity and sensitivity of CTLpf assay. We are in agreement with others that the Eu-release assay is specific and re- producible. It can be used for the CTLpf estimation as well as other T cell and non-T cell cytotoxicity assays.

Keywords: europium-release; cytotoxicity; transplantation; fluorometry; recombinant interleukin 2; T lym- phocytes.ocytes.
Stimulatory Effect of Ovine Colostrinine (a Proline-Rich Polypeptide) on Interferons and Tumor Necrosis Factor Production by Murine Resident Peritoneal Cells (Short Communication).
ZOFIA BLACH-OLSZEWSKA 1 and MARIA JANUSZ 2

Abstract. We describe effects of ovine colostrinine (proline-rich polypeptide – PRP) isolated from ovine colostrum and nonapeptide fragment of PRP on interferon (IFN) and tumor necrosis factor (TNF) production by murine resident peritoneal cells (RPC). The cells from several mouse strains have been found to produce small amounts of IFN-ß and TNF-a constitutively. The colostrinine at concentrations of 1-100 ixg per one ml of cell suspension containing 1 x 106 RPC isolated from BALB/c mice, enhanced the IFN and TNF production by 3 – 30 folds. Upregulation of TNF and IFN production has been observed in the RPC cultures that produced spontaneously less than 16 units of the cytokines only. Synthetic nonapeptide fragment of the colostrinine (Val-Glu-Ser-Tyr-Val-Pro-Leu-Phe-Pro) at concentration of 1-100 gg/ml stimulated TNF syn- thesis but not IFN production. In 1996 Inglot et al. suggested that the colostrinines may be classifed as cytokines produced by the mammary gland of mammals. In this paper we have found that the ovine colost- rinine at low concentrations modulate the production of other cytokines (IFN-ß and TNF-a) in mouse cells that means that it may function in the cytokine network.

Keywords: ovine colostrinine; murine resident peritoneal cells; interferons; tumor necrosis factors.actors.
Interleukin 12 and Direct Cytotoxicity of Spleen Lymphocytes to Listeria innocua-Phagocyting Syngeneic Macrophages in C57BL/6 and BALB/c Mice.
JACEK SZELIGA, DANUTA FIUK, MARIA WALENCKA and TERESA GOŚCICKA

Abstract. Production of interleukin 12 (IL-12) during cytotoxic reaction of CS7BL/6 and BALB/c spleen lymphocytes or their subpopulations: T+B, T, CD4+ and CD8+ cells to L. innocua-phagocyting syngeneic macrophages was examined. The effector cell donors were untreated or L. innocua-infected. The number of surviving bacteria in phagocytes was tested and IL-12 level in culture supernatants of reacting cells was determined. CS7BL/6 mice, resistant to Listeria infection, were found to develop stronger cell cytotoxicity to bacteria-phagocyting syngeneic macrophages than BALB/c mice. The lymphocytes responsible for that pheno- menon were of CD8+ phenotype. IL-12 was produced only during nonspecific cytotoxic reaction of CD4+ and CD8+ T cells. It is suggested that the innate resistance of mice to Listeria is dependent on their ability to develop a specific cytotoxic reaction and IL-12 production.

Keywords: interleukin 12; cytotoxicity; phagocyting macrophages; Listeria.ges; Listeria.
Sensitivity of Human Cutaneous Mast Cells to Anaphylactic and Nonimmunological Stimuli.
EWA BRZEZIIVSKA-BLASZCZYK1 and ANNA ZALEWSKA2

Abstract. . Mast cells represent significant cellular element of the skin. It is now postulated that they play an important role in cutaneous homeostasis and are engaged in some pathological processes as well. The aim of our study was to examine sensitivity of human cutaneous mast cells to anaphylactic and anaphylactoid stimuli. The studies were performed on human cutaneous mast cells obtained from healthy skin by enzymatic dispersion technique. The mast cells were activated in vitro with anti-IgE, concanavalin A (Con A), compound 48/80, substance P (SP) or tumor necrosis factor a (TNF-a). We have noticed that skin mast cells were susceptible to Presentation the challenge with anti-IgE and Con A, and histamine release was dose- and time-dependent. In both cases histamine release was high (44.0±4.1% with anti-IgE at dilution I:500 and 20.1 ±2.4% with Con A in concent- ration 500 qg/ml). Cutaneous mast cells were challenged in a dose- and time-related fashion with compound 48/80, however histamine release was low (9.8±2.4%, at concentration of compound 48/80 – 100 ltg/ml). SP and TNF-a also activated mast cells but the magnitude of histamine release was not high (up to 7.1 ±0.9%, SP in concentration 10 4 M and 17.4 ± l.1 %, TNF-a in concentration 10 6 M) and maximal after 20 min reaction.

Keywords: cutaneous mast cells; anaphylactic histamine release; anaphylactoid histamine release; substance P; tumor necrosis factor aactor a

Experimental and Clinical Therapy

Interferon-Inhibiting Activities in Sera of Patients with Lung Cancer (Short Communication).
KONSTANTINOS KARMANIOLAS, GEORGE TSANTAKIS and MARIA HAVREDAKI

Abstract. A differential pattern of sera exhibiting interferon-inhibiting activity in cases of histologically confirmed lung cancer patients have been observed. The sera distribution was dependent on the clinical entity and the cell line specifďcity used measuring IFN-inhibition. Although either small cell lung cancer (SCLC) or non small cell lung cancer (NSCLC) sera exhibiting in a high percentage (80 and 75% respectively) such an activity, in any of the 3 lines tested, the ratio of sera exerting inhibition in each cell line was different. Such cell specific systems could be of prognostic/predicting value for effective therapeutic schedules of specific clinical entities.

Keywords: interferon-inhibiting activity; serum inhibitors; lung cancer patients.tients.
Fffect of Bacterial Extract, IRS-I9, on the Concentration of Hydrogen Peroxide and Myeloperoxidase Activity in Nasal Washings of Patients with Chronic Bronchitis.
DARIUSZ NOWAK 1, MICHAŁ, PROZYIVSKI Z, TADEUSZ PIETRAS 1, ROBERT STOLAREK 1, PIOTR MAZERANT 3 and MICHAL LEDER 3

Abstract. Twenty eight adult patients of both sexes with chronic bronchitis participated in an open study to determine the eh’ect of intranasal treatment with IRS-19, an immunomodulating agent, on the number of polymorphonuclear leukocytes (PMNL), HzOz concentration and myeloperoxidase (MPO) activity in nasal washings. The number of PMNL recovered from nasal spaces increased from 4460±3960 to 10490± 10950 cells/ml (p < 0.02) after two month administration of IRS-19. It was accompanied by 2.6- and 1.4-fold increase (p < 0.001) in MPO activity and HzOz concentration, respectively. However, no correlation was found betwe- en increments in these three variables. Since PMNL and MPO – HzOz – CI system are involved in the first line of defense against invading pathogens it is suggested that above mentioned changes may represent one among mechanisms leading to enhancement of antibacterial defence in the airways in response to treatment with IRS-19.

Keywords: chronic bronchitis; bacterial extract; IRS-19; nasal washings; myeloperoxidase; hydrogen peroxi- de; polymorphonuclear leukocytes.ar leukocytes.
Urinary Gamma-Glutamyl Transferase Activity in Rats with Nonsteroidal Anti-Inflammatory Drug-Induced Nephrotoxicity (Short Communication).
SÜKRAN KOCAOGLU 1, AYSEN KARAN 1, TAYFUN BERKAN 2, GÜLÇIN BASDEMIR 3 and RIYAT AKPINAR 2

Abstract. Excretion of urinary gamma-glutamyl transferase (GGT) was studied in rats following p.o. ap- plication of high doses (10 mg/kg/day) of indomethacin, diclofenac sodium or piroxicam for 28 days. Measure- ments of 24 h urinary GGT activity and urinary creatinine were carried out on 29th day. Histological examinations of kidneys were performed on day 29. The mean value for urinary GGT was found to be 0.77±0.05 U/mg creatinine (n =16) in the control group. The mean activities in the treated groups were as follows: 1.30 ± 0.15 U/mg creatinine (n =17, indomethacin); 1.22 ± 0.25 U/mg creatinine (n = 4, diclofenac); 1.54±0.39 U/mg creatinine (n = 5, piroxicam). The mean enzyme activities in indomethacin and piroxicam treated groups were significantly higher than in the control group (p < 0.02 and p < 0.03, respectively), while no significant dill’erence has been found between the group treated with diclofenac and control group (p 0.05). Histological examinations of renal tissues of indomethacin, piroxicam and diclofenac treated groups showed minimal glomerular abnormalities. Thus, determination of urinary GGT may be useful to investigate the nonsteroidal anti-inflammatory drugs (NSAIDs) related renal toxicity in rats.

Keywords: nephrotoxicity; gamma-glutamyl transferase; indomethacin; diclofenac; piroxicam.oxicam.
Influence of Mesna on Urotoxic Effects of Selected Bromosubstituted Analogs of Ifosfamide.
HALINA KUŚNIERCZYK 1, LESZEK KONARSKI 1, PRZEMYSŁAW KOWALSKI2 and CZESŁAW RADZIKOWSKI 1

Abstract. Bromofosfamides, the group of novel compounds closely related to ifosfamide, are currently in the stage of advanced preclinical evaluation. Ifosfamide, although itself the effective antineoplastic drug useful in situations which have proved refractory to cyclophosphamide therapy, has the side-effect toxicities caused by its metabolites that pose clinically a very real problem. One of their manifestations is the severe urinary tract toxicity which now could be adequately managed by conjunctive administration of mesna (sodium 2-mercap- toethane sulphonate). In this study we have compared the magnitude of urotoxic effects elicited by ifosfamide and two bromofosfamide compounds – racemate and S(-) isomer of chlorobromofosfamide (ClBrs) – selec- ted previously on the base of their superior antitumor activity in advanced animal tumor models. The urotoxic effects, expressed by the increase of urinary bladder weight and histopathologically defined organ wall edema, were estimated in healthy mice 24 h following single intraperitoneal or oral administrations of tested com- pounds which were applied in amounts equal to curative, sublethal or lethal doses. It was found that the expression of toxic effects revealed by both ClBrs was statistically significantly lower as compared to ifos- famide. Mesna coadministration prevented urotoxic effects almost completely in mice treated with ifosfamide or racemic ClBr. Somewhat lower efflcacy of uroprotection with mesna was observed in the case of S( – ) isomer of ClBr.

Keywords: ifosfamide; bromofosfamides; urotoxicity; mesna uroprotection.uroprotection.
Vitamin C Inhibits Random Migration of Malignant Pleural Effusion Mononuclear Cells.
LJUBICA PROKIC 1, IVANKA VILIĆ l, LJILJANA VUCKOVIĆ-DEKIĆ 2, ZORA NESKOVIĆ-KONSTATINOVIĆ 2 and SNEZANA SUSNJAR 2

Abstract. Although many epidemiological studies indicate protective effect of vitamin C against a variety of human malignancies its mechanism(s) of action is questionable. The presented results show that the part of its effect may be accomplished by mononuclear cells, as necessary participants in body defence. Namely, in a long-term in vitro assay we tested vitamin C influence on random migration ability of malignant pleural effusion mononuclears (PEM) obtained from breast cancer patients. Vitamin C in a dose- (50-500 g) and time-dependent (4-44 h) manner inhibited PEM motility, suggesting that immobilization of cells in situ may contribute to its beneficial effect in human cancers.

Keywords: malignant pleural effusion mononuclears; random migration; vitamin C.on; vitamin C.

Various

Effect of Suramin on Human Interferon alpha Binding to Cell Receptors.
MARTYNA KANDEFER-SZERSZEŃ 1, KRISTINA RUUTH 2 and ERIK LUNDGREN 2

Abstract. Radioiodinated human recombinant interferon a88 (l2sl-HuIFN-a88) binds to high affinity recep- tors of IFN sensitive Daudi cells (Burkitt lymphoma cell line). Suramin, a low molecular weight (1429), polyanionic compound at concentrations 105 -175 ItM completely abolished 125I-IFN-a88 binding to Daudi cells at low temperature (4oC). At 37oC, however, its effect was only partial and depended on incubation time. Suramin also dissociated IFN-a88-receptor complexes but, upon incubation of cells with IFN at 37C IFN-receptor complexes, became gradually less sensitive tu suramin action. Dissociation by suramin IFN-a88-receptor complexes prevented induction of (2-5)A synthetase activity and inhibited down-regula- tion of IFN receptors on Daudi cells. We suppose that the first reaction which represents IFN binding to the surface receptors is inhibited and dissociated by suramin, but when IFN is transferred to a tight activation complexes on the cell membrane or internalized, such complexes cannot be dissociated by suramin.

Keywords: interferon a receptors; suramin; (2-5)A synthetase.)A synthetase.
Pathogenesis of Breast Carcinoma. Immunohistochemical Study.
ALBINA ŻÓŁTOWSKA

Abstract. Synaptophysin and/or chromogranin A may be expressed in epithelial cells of normal and dysplastic mammary gland and in some cancer of the breast. This work indicates that some stromal cells form thin walled vascular channels and from their perivascular mesenchyma arise myoid-like cells, some with cross-like strations. These myoid-like cells have been stained with antibody to smooth muscle actin, rarely to desmin and sarcomeric actin as well as are strongly positive for synaptophysin and/or chromogranin A. From those cells arise cancer cells. Some cancer cells also expressed desmin, sarcomeric actin or smooth muscle actin. Because synaptophysin positive are neuromuscular endplates, it is a question whether the presence of synaptophysin in the progenitor of neoplastic cells of myogenic origin is due to the synaptic dysfunction in molecular mecha- nism of the epithelial-parenchyma and mesenchymal stroma interaction. The location of S100-protein positive dendritic cells distributed in regular pattern in suprabasal layer of the mammary gland indicate that these cells may arise from preadipocytes which take part in morphogenesis of the breast. All cases of the breast cancer demonstrated thymosin al, some of them also showed Hassall’s-like bodies, mucin secretion and minute calcification. Whether the interaction of epithelial cells and S100-protein positive dendritic cells in tissue other than the thymus has similar dynamic activity to that of Hassall’s bodies of the thymus remains to be determined. Both, epithelial cells of Hassall’s bodies and epithelial cells of the investigated breast are of myogenic origin. The above consideration suggests that the cells forming vascular channels are with a differen- tiation defect as they are created in altered stromal mesenchyma and the presence of thymic growth factors might induce tumor growth.

Keywords: breast cancer cells; synaptophysin; S100-protein; dendritic cells; preadipocytes; thymosin al.; thymosin al.
Coincidence between Spontaneous Release of Interferon and Tumor Necrosis Factor by Colostral Leukocytes and the Production of a Colostrinine by Human Mammary Gland after Normal Delivery.
EGBERT PIASECKI l, ANNA D. INGLOT l, MAŁGORZATA WINIARSKA 3, KATARZYNA KRUKOWSKA l, MARIA,IANUSZ 2 and JÓZEF LISOWSKI 2

Abstract. Synaptophysin and/or chromogranin A may be expressed in epithelial cells of normal and dysplastic mammary gland and in some cancer of the breast. This work indicates that some stromal cells form thin walled vascular channels and from their perivascular mesenchyma arise myoid-like cells, some with cross-like strations. These myoid-like cells have been stained with antibody to smooth muscle actin, rarely to desmin and sarcomeric actin as well as are strongly positive for synaptophysin and/or chromogranin A. From those cells arise cancer cells. Some cancer cells also expressed desmin, sarcomeric actin or smooth muscle actin. Because synaptophysin positive are neuromuscular endplates, it is a question whether the presence of synaptophysin in the progenitor of neoplastic cells of myogenic origin is due to the synaptic dysfunction in molecular mecha- nism of the epithelial-parenchyma and mesenchymal stroma interaction. The location of S100-protein positive dendritic cells distributed in regular pattern in suprabasal layer of the mammary gland indicate that these cells may arise from preadipocytes which take part in morphogenesis of the breast. All cases of the breast cancer demonstrated thymosin al, some of them also showed Hassall’s-like bodies, mucin secretion and minute calcification. Whether the interaction of epithelial cells and S100-protein positive dendritic cells in tissue other than the thymus has similar dynamic activity to that of Hassall’s bodies of the thymus remains to be determined. Both, epithelial cells of Hassall’s bodies and epithelial cells of the investigated breast are of myogenic origin. The above consideration suggests that the cells forming vascular channels are with a differen- tiation defect as they are created in altered stromal mesenchyma and the presence of thymic growth factors might induce tumor growth.

Keywords: breast cancer cells; synaptophysin; S100-protein; dendritic cells; preadipocytes; thymosin al.; thymosin al.

Vol. 45, No. 2-3, 1997

Review

Is the Prion Structure Solved?
PAWEL P. LIBERSKI and JOLANTA BRATOSIEWICZ-ZAPART

Abstract. We report here on the current knowledge on the nature of the scrapie agent or prion. Several lines of evidence suggest that the abnormal isoform of prion protein (PrP) is crucial for scrapie infectivity while evidence that PrP is also a part of the entire particle of the scrapie agent (prion) is much weaker. There is no doubt, however, that conformational changes (transitions from a-helical into ß-pleated structures) of PrP underlay scrapie pathogenesis. In view of the notorious puzzling nature of the scrapie agent, the electron microscopic search for the ultrastructural correlate of it is still warranted. Thus, we discuss the nature of tubulovesicular structures (TVS), the only diseases-specifďc particles known so far and the association between TVS and PrP fibrils which was recently discovered.

Keywords: transmissible spongiform encephalopathies; prion protein (PrP), pathological isoform of PrP; mad cow disease; scrapie.ease; scrapie.
Immunological Adjuvants and Their Modes of Action.
ANTHONY C. ALLISON

Abstract. New adjuvant formulations contain a vehicle, which carries antigens to antigen-presenting cells. Examples of vehicles are liposomes, immune-stimulating complexes and microfluidized squalene-in-water emulsions. Adjuvant formulations may contain immunomodulators, which augment cytokine production, such as a synthetic muramyl dipeptide analog or monophosphoryl lipid A. In a primary cascade of cytokine production at the site of antigen + adjuvant injection, TNF-a promotes the migration of dendritic cells (DC) to lymphoid tissus while GM-CSF accelerates the differentiation of DC into efficient presenters of antigens to T cells. Adjuvants also up-regulate a secondary cascade of cytokines in lymphoid tissues responding to antigenic stimulation: IL-12 augments the production of IFN-y, which favors the production of antibodies of protective isotypes (IgG2a in the mouse). Thus adjuvants can regulate immune responses qualitatively as well as quantitatively. Adjuvant formulations can also activate complement, generating C3d, which binds CD21 on follicular dendritic cells (FDC) and B cells. FDC targeting favors the generation of B lymphocyte memory, which is important for vaccination.

Keywords: adjuvant; dendritic cells; follicular dendritic cells; squalene emulsions; cytokines.okines.
Molluscan Immune Defenses.
ZDZISLAW GLIŃSKI 1 and JAN JAROSZ 2

Abstract. The interest of marine invertebrates as food resources provides a major interest to study molluscan immunity for better understanding of the host response to pathogens. Molluscs possess a natural immunity formed by anatomical and chemical protective barriers that prevent damage of the underlying tissues, body fluid losses and the infections of pathogenic microorganisms and parasites. The main physical barrier is shell and mucus which cover the soft body of molluscs. The integrity of body coverings is supported by blood clotting and wound healing. The internal defense mechanisms of molluscs involve such cellular reactions as: phagocytosis, nodule formation, encapsulation, pearl formation, atrophy, necrosis and tissue liquefaction. Granular hemocytes are the most numerous cell type of molluscan blood active in cellular defenses. Invaders small in size are eliminated by phagocytosis in which participate lectins and products of prophenyloxidase system activation. Numerous and large intruders are eliminated by nodule formation or encapsulation, either cellular or humoral. Humoral components of molluscan immunity are formed by lysozyme activity, lectins and the phenyloxidase system. Up to now the role of mercenenes, paolins, acute phase reactants, a2-macro- globulins and multifunctional binding proteins with anti-protease activity is not well clarified yet. Research prospects on the field of molluscan immunology should essentially be devoted to study cellular defense functions and humoral elfectors to select pathogen-resistant molluscs. This aim could also be achieved by the identification and characterization of immune genes which are candidates for molluscs genetic transformation.

Keywords: molluscan immunity; mucus barriers; encapsulation and nodule formation; pearl formation; paolins; mercenenes.s; mercenenes.
Rhythms of Immunity.
BARBARA PŁYTYCZ and ROLF SELJELID

Abstract. Rhythms of daily activity are found in all vertebrate species, some of them being diurnal (like humans, dogs, pigeons), others – nocturnal (like mice, rats and bats). Some species undergo very pronounced seasonal changes, as they hibernate in the winter or mate only at the specific seasons. The main regulator (a clock and a calendar) for daily and seasonal rhythms is the periodicity of the external light-darkness, reflected by the periodicity of melatonin secretion from the pineal gland, which is inhibited by light and induced during the darkness. In contrast to melatonin which peaks during the night both in diurnal and noctural species, the cyclicity of other hormones and several immune parameters correlates with the pattern of the animal locomo- tor activity-resting. The immune parameter that peaks at one time of day for a diurnal species peaks about 12 h later for a nocturnal one. Various immune parameters peak at various time points, anticipating an encounter with pathogens during the period of activity while energetically expensive resolution of the immune response during the resting. Daily and seasonal cyclicity of the immune functions are temporally integrated with other physiologic and behavioral processes and all of them are regulated and coordinated with daily and seasonal changes of an external environment by the neuroendocrine homeostatic system.

Keywords: daily rhythms; seasonal rhythms; immunity; immune system; melatonin; adaptation.n; adaptation.
Regulation of Cytokine Production by Eicosanoids and Nitric Oxide.
JANUSZ MARCINKIEWICZ

Abstract. Cytokines are widely regarded as regulatory molecules of inflammatory and immune reactions. Nevertheless, the details of functioning of the complex cytokine network are not yet fully understood. Recent data indicate that eicosanoids, primarily the products of the inducible form of cyclooxygenase (COX-2), are involved in the regulation of cytokine production. We have shown that prostaglandins of E series are no longer only suppressor molecules but they selectively up- or down-regulate the cytokine production. Similarly, nitric oxide (NO) generated in activated immune cells by inducible isoform of nitric oxide synthase (iNOS), is considered to be an immunoregulatory molecule. In this article we present a new concept of interactions between cytokines, eicosanoids (prostaglandins and leukotrienes) and NO. Finally, the impact of these molecu- les on the regulation of the immune system is discussed.

Keywords: immunoregulation; cytokines; eicosanoids; prostaglandins; leukotrienes; nitric oxide. oxide.

Clinical Immunology

Immunological Status of Septic and Trauma Patients. I. High Tumor Necrosis Factor a Serum Levels in Septic and Trauma Patients Are Not Responsible for Increased Mortality; a Prognostic Value of Serum Interleukin 6.
BARBARA ADAMIK, MICHAŁ ZIMECKI, ANDRZEJ WŁASZCZYK and ANDRZEJ KÜBLER

Abstract. The aim of this study was to monitor plasma interleukin 6 (IL-6), and tumor necrosis factor a (TNF-a), levels in patients with sepsis, septic shock and multiple organ dysfunction syndrome admitted to the intensive care unit. The patients obtained adequate supportive therapy. Plasma samples were taken upon admission, then on days 1, 2, and 5 following admission. IL-6 and TNF-a levels were determined using bioassays (7TD1 and WEHI-164.13 indicator cell lines, respectively). The results showed that the kinetics of the cytokine release in septic patients differed significantly between survivors and nonsurvivors. In survivors IL-6 concentrations were initially high, fell down rapidly on day 1 after admission, and persisted very low throughout the monitoring time. In contrast, relatively low IL-6 levels in the nonsurvivors, registered upon admission, rose significantly with peak values on day 3 of observation, declining thereafter. TNF-a levels were initially higher in survivors than in nonsurvivors, declined on day 1 following admission, and on day 5 they were higher than the initial values. In nonsurvivors, on the other hand, the starting concentrations of TNF-a were much lower than in survivors with a peak on day 3 with a tendency to fall on day 7. The profiles of cytokine production by traumatic patients (90a/o survivors) revealed low and progressively diminishing levels of IL-6, contrasting with constantly increasing concentratios of TNF-a within the monitoring period. We conclude that high IL-6 levels in septic patients accompanied by high TNF-a levels may indicate bad prognosis. In contrast, rapidly diminishing serum IL-6 levels, even in the presence of high TNF-a levels, could indicate a very good chance for survival. Similar conclusion can be drawn from the monitoring of cytokine production in traumatic, nonseptic patients since almost all of them recovered. We also speculate that TNF-a presence in circulating blood is essential for regeneration of tissues and wound healing.

Keywords: sepsis; trauma; interleukin 6; tumor necrosis factor a.ctor a.
Modulating Effect of Interleukin 2 Therapy on Interferon Production by Blood Leukocytes of Patients with Minimal Residual Hematological Disease.
MARTYNA KANDEFER-SZERSZEŃ l, WOJCIECH LEGIEĆ 2, ANNA DMOSZYŃSKA2 and AGNIESZKA SZUSTER-CIESIELSKA 1

Abstract. This study was designed to investigate the effect of 1.8 x 106 U/day interleukin 2 (IL-2) therapy on interferon (IFN) production. Patients enrolled in the study suffered from multiple myeloma (MM), Hodgkin’s disease (HD) and non-Hodgkin lymphoma (NHL). All of them were in remission after chemotherapy or radiotherapy. Results indicated that IL-2 given subcutaneously at a dose of 1.8 x 106 U/day for 3 weeks induced IFN-y in serum of patients and caused a prolonged effect on the ability of blood leukocytes to produce IFN-y after stimulation in vitro by mitogen phytohemagglutinin (PHA). Such enhancement of IFN-y production may be beneficial for antitumor immune response. Low-dose IL-2 therapy was well tolerated by all patients and side effects not exceeding II grade of toxicity according to WHO scale were observed. Five patients with MM have relapsed 3-10 months after cesation of IL-2 therapy but 15 patients 18 months after tberapy were in complete remission.

Keywords: interleukin 2 low-dose therapy; hematological disease; interferon.e; interferon.
Isoprinosine and Levamisole as Stimulators of Interferon Production in Blood Leukocytes of Patients with Alcoholic Liver Cirrhosis.
JADWIGA DANILUK 1 and MARTYNA KANDEFER-SZERSZEŃ 2

Abstract. Blood leukocytes of 16 patients with alcoholic liver cirrhosis and 18 healthy controls were induced for interferon (IFN) production by phytohemagglutinin (PHA) and concanavalin A (ConA) in the presence or absence of isoprinosine and levamisole at concentrations of 10 pg/ml and 1 ng/ml. This interferon was neutralized in 87-95% by anti-HuIFN-y monoclonal antibodies. In the presence of the drugs the IFN-y production was enhanced, however, IFN-y titers yielded from leukocytes of cirrhotic patients were still below the titers observed in stimulated and unstimulated blood leukocytes of healthy controls. For example, IFN titers induced by PHA in the presence of levamisole (1 ng/ml) in cirrhotic patients were 2.5 times lower (20.2±11.1 U/ml) in comparison to healthy subjects (50.6±27.3 U/ml).

Keywords: isoprinosine; levamisole; interferon y; alcoholic livčr cirrhosis.včr cirrhosis.
Changes in the Phagocytic Cells in Children Treated with Continuous Ambulatory Peritoneal Dialysis.
MARIA WĄSIK, MARIA BLAIM, DANUTA KOLEWSKA, EWELINA JANOTA-KRAWCZYK, MAŁGORZATA TOMASZEWSKA-PAŃCZYK and MARIA SIENIAWSKA

Abstract. Children on continuous ambulatory peritoneal dialysis (CAPD) in endstage renal failure are highly exposed to peritonitis. Peritoneal macrophages (PM) and blood neutrophils (PMNC) are the first line of defense against invading microbes. This study was undertaken for assessing surface receptors expression on PM and PMNC and to check their ability to phagocytosis and killing of bacteria. We have found that in spite of the decreased number of PM in dialysate fluid their viability and activity significantly increased during CAPD. Moreover, higher number of PM expressed CDl6 and CD3S antigens (FcRIII and C3bR, respectively) in comparison with the results observed at CAPD onset. The number of PMNC expressed of these two antigens in uremic children blood were significantly lower in compararison with healthy control. The number of CDl6 positive cells increased under influence of CAPD only temporaly. CAPD caused improvement of phagocytosis and intracellular killing of bacteria by PM but not by PMNC. There is discus- sed here influence of uremia and CAPD on surface antigens, function of phagocytes as well as renewal of PM during CAPD.

Keywords: peritoneal dialysis; phagocytosis; marcophages; blood neutrophils.ophils.
Eosinophil Cationic Protein in Serum of Corticosteroid-Sensitive and Resistant Bronchial Asthma.
JANUSZ PATKOWSKI, KRZYSZTOF WYTRYCHOWSKI and WOJCIECH MĘDRALA

Abstract. Theories for the inflammatory basis of bronchial asthma are presented. The phenomenon of corticosteroid resistance (CR) in bronchial asthma is also discussed. Resistance to corticosteroids, which occurs in about 5% patients with moderate and severe asthma, presents still an important diagnostic and therapeutical problem. In addition, present opinions on the role of eosinophils in the allergic bronchial inflammation were analyzed. The aim of this study was to monitor serum eosinophil cationic protein (ECP) level in asthma patients, sensitive and resistant to glucocorticosteroids (GCS), before and after prednisolone treatment. The resistance to steroids was determined, based on the oral prednisolone test according to Carmichael and vasoconstriction assay according to Stoughton and McKenzie. In the group of cortico- steroid-sensitive (CS) asthmatic patients a statistically significant decrease of ECP level was observed, after 10 day administration of prednisolone in a daily dose of 20 mg, which was associated with a meaningfull increase of FEV, value. On the other hand, the level of ECP in the serum of patients resistant to cortico- steroids, although also decreased under influence of prednisolone, was not correlated with the increase of FEV1 value.

Keywords: corticosteroid-resistant and -sensitive bronchial asthma; eosinophil cationic protein.ionic protein.
Immunological Status of Patients Subjected to Cardiac Surgery: Effect of Lactoferrin on Proliferation and Production of Interleukin 6 and Tumor Necrosis Factor a by Peripheral Blood Mononuclear Cells in Vitro.
ANDRZEJ WŁASZCZYK, MICHAŁ ZIMECKI, BARBARA ADAMIK, GRAŻYNA DUREK and ANDRZEJ KÜBLER

Abstract. The aim of this investigation was to monitor proliferation and cytokine production by peripheral blood mononuclear cells (PBMC) of patients subjected to cardiac surgery. Another goal was to establish regulatory effects of lactoferrin (LF) on these immune reactions in uitro. PBMC were tested before, during surrgery and on day 1 and day 8 -10 following operation. In control donors, low spontaneous and phytohe- magglutinin (PHA)-induced proliferation of PBMC, as well as lipopolysaccharide (LPS)-induced TNF-a secretion was stimulated by LF, but high production of this cytokine was inhibited. In patients, the prolifera- tion of PBMC and the ability to produce IL-6 TNF-a by these cells underwent characteristic changes ;depending on preoperative immune reactivity of patients. In general, low, preoperative reactivity of PBMC showed a tendency to increase within the monitoring period whereas moderate/high responsiveness was diminishing. LF had, in majority of cases, a down-regulatory effect on the proliferative response, best pro- nounced in patients of high/moderate preoperative response. Similarly, LF exhibited, in general, an inhibitory effect on LPS-induced IL-6 production. In terms of TNF-a production, a considerable up-regulatory effect of LF, particularly in low responding patients was of a special interest. In summary, we suggest that LF may play a role in lowering the immune response of patients to surgery and promoting tissue regeneration.

Keywords: cardiac surgery; peripheral blood leukocytes; proliferation; interleukin 6; tumor necrosis factor a; lactoferrin.ferrin.
Release of Tumor Necrosis Factor a and Its Soluble Receptor p55 in Clinical Septic Syndrome
BOGDAN MODZELEWSKI

Abstract. Tumor necrosis factor a (TNF-a, cachectine) attracts much clinical interest when it looses its primary immune properties and, in the septic syndrome, initiates its destructive activity against host tissues. Behaviour of TNF-a and of p55, its soluble receptor (sTNF-R p55), was studied in 50 patients with septic syndrome. Their clinical state was assessed by means of the APACHE II score. Group I consisted of 23 patients who had survived the infection. Twenty seven patients (54%) died, constituting group II. The concentrations of TNF-a and sTNF-R p55 were determined according to the ELISA method. The obtained results were referred to the severity of the patients’ clinical state, their survival rate, the number of lactates and the quantity of the C-reactive protein in blood and the number of blood platelets and neutrophils. It has been documented that in septic syndrome patients with poor prognosis the concentration of TNF-a rises, together with a similar decrease of sTNF-R p55 concentration. This relationship is very important for the prognosis in septic syn- drome.

Keywords: cytokines; tumor necrosis factor; soluble receptor p55; sepsis; septic syndrome.ndrome.
Distribution of Apolipoprotein E4 Isoform in Alzheimer’s Disease in Poland (Short Communication).
JOLANTA BRATOSIEWICZ-ZAPART, IWONA KŁOSZEWSKA, TOMASZ SOBÓW and PAWEL P. LIBERSKI

Abstract. Alzheimer’s disease (AD) is the most common human progressive dementia linked, in its sporadic form, to a common polymorphism within a gene for apolipoprotein E (APOE) mapped to chromosome 19. Individuals with APOE 4 isoforms are more prone to develop sporadic from of AD than those who carry APOE 2 or APOE 3 alleles. As distribution of APOE isoforms in Poland is unknown, we decided to study it in AD and control cases. In AD group, three patients (23.la/o) were homozygous for APOE 4, four cases (30.8%) were heterozygous and six cases (46%) carried no APOE 4 allele. In control group (n-11), only one case (9.1%) was homozygous for APOE 4 allele, no case was heterozygous for APOE 4 while 10 cases (90.9%) carried no APOE 4 alleles. Our results are virtually identical to those reported from Western countries.

Keywords: apolipoprotein E; Alzheimer’s disease.isease.

Immunochemistry

Haptoglobin Concentration in Blood Sera of Hoofed Mammals in Captivity.
TADEUSZ STEFANIAK, ANIELA GOSPODAREK, FRANZ KLIMA, DIETER SCHADOW and IWONA KĄTNIK

Abstract. There were 151 serum samples of the zoological species of hoofed animals from the Bovidae, Cervidae and Equidae families examined for the presence of haptoglobin (Hp). In the majority of samples the Hp levels were close to the ones defined in the related species of domestic animals (86.49, 52.38 and 69.7% in particular families, respectivel . In females (13.94% in the Bovidae family, 63.64% in the Cervidae,15o/o in the Equidae) more often than in males (11.11, 26.32 and Oo/o, respectively) the increased Hp concentrations were found. What is striking it is the relatively high frequency of the raised Hp concentrations occurrence in the Cervidae sera, which may be connected with the higher responsiveness to the factors inducing Hp rise in this family. The contribution of infectious, parasitic, stress-inducing factors and injuries is taken into consideration as the main cause of the raised levels occurrence in the sera of animals kept in zoological gardens. The obtained results suggest the potential usefulness of the Hp estimation as a routine examination in the diagnostics of the intlammation in wild species of Bovidae and Cervidae as well as in monitoring the animal well being in the zoological gardens.

Keywords: haptoglobin; wild Bovidae; wild Cervidae; wild Equidae; zoological garden.garden.
Some Parameters Influencing Immunoassay of Human and Horse Myoglobins
IWONA E. KOCHANOWSKA, MARIANNA KUROPATWA and APOLINARY SZEWCZUK

Abstract. It was noted that human and horse sera as well as human heart and skeletal muscle homogenates or extracts distinctly decrease immunoassays of purified myoglobins. The assays of homogenate and extract myoglobins could be many times increased by precipitation certain proteins with concentrated ammonium sulfate or sodium chloride. Also in homogenates and extracts incubated for several days increased assays of myoglobins were noted. The obtained results indicate that both myoglobins occur in complex with other tissue component(s).

Keywords: human myoglobin; horse myoglobin; immunoassay of myoglobins; polyclonal and monoclonal antibodies.bodies.
Mechanism of Antigenic Variation in Shigella flexneri Bacilli. IV. Role of Lipopolysaccharides and Their Components in the Sensitivity of Shigella ,flexneri lb and Its Lac+ Recom binant to Killing Action of Serum.
WŁODZIMIERZ DOROSZKIEWICZ

Abstract. The effect of lipopolysaccharides (LPS) on the normal bovine serum (NBS) bactericidal reactions against mixture of S. flexneri 6713 lb strain and its 3b Lac+ recombinant were investigated. The serum killing of S. flexneri strains was inhibited, in different degree, by LPS extracted from either organisms. These proper- ties were mainly due to LPS molecules; the lipid A fraction showed only low anticomplement activity, the polysaccharide fraction inhibited the killing activity of NBS in very low degree even at high concentration. These studies suggest that LPS composition especially the O-antigen polysaccharide chain contributes to the susceptibility of S. flexneri strains to NBS bactericidal activity.

Keywords: Shigella flexneri; complement; lipid A; lipopolysaccharide.olysaccharide.

Various

Effect of Viral Infections on the Ability of Human Endothelium to Interferon, Tumor Necrosis Factor and Interleukin 6 Production.
EWA ZACZYŃSKA, ZOFIA BŁACH-OLSZEWSKA, JOLANTA SENDER and WIKTOR JAROSZ

Abstract. We have reported that cultured human umbilical cord vein endothelial cells (HUVEC) differ from endothelium present on vein surface of organ culture (OC) in production of cytokines and susceptibility to viral infections. In this paper we present the effect of viral infections on interferon (IFN), tumor necrosis factor (TNF), and interleukin 6 (IL-6) production in two culture systems: HUVEC and OC. Infection of 24-48 h HUVEC with herpes simplex type 1 (HSV-1) and vesicular stomatitis virus (VSV) reduced the amounts of IL-6 and TNF produced in comparison to those released spontaneously by uninfected cells. No IFN was detected in media from infected and uninfected HUVEC. Limited viral infections of 3-h-HUVEC and OC usually diminished their efflciency of IL-6 and TNF production. In the case of IL-6 synthesis by OC, effect of viral infection depended, however, on the constitutive synthesis of the cytokine. When spontaneous produc- tion was high ( 800 U/ml), VSV and HSV-1 infection reduced IL-6 level by 2-50 times; in the case of low production (<150 U/ml) the stimulation effect (2-4 fold) was observed. OC released spontaneously some IFN activity (2-32 U/ml). HSV-1 infection of OC reduced IFN level, while VSV in single cases slightly upregulated IFN synthesis.

Keywords: human endothelium; virus infections; interferon; tumor necrosis factor; interleukin 6.ukin 6.
Metoclopramide and Nitrendipine Are Unable to Enhance Cytotoxicity of Mitoxantrone in Sensitive and Resistant Murine Leukemias in Vitro.
EWA ZACZYŃSKA, ZOFIA BŁACH-OLSZEWSKA, JOLANTA SENDER and WIKTOR JAROSZ

Abstract. The influence of metoclopramide and nitrendipine on the antitumor action of mitoxantrone (MX) in murine leukemias L1210 and P388 was investigated. These agents were administered in combination with MX to mice bearing sensitive or resistant leukemias. Mitoxantrone resistant P388 was developed ivI tii;o by repeated passages of the tumor to animals treated with suboptimal dose of the drug. Survival time of mice receiving combined therapy were not prolonged as compared with animals treated with MX alone. Both metoclopramide and nitrendipine were unable to overcome MX resistance even partially.

Keywords: mitoxantrone resistance; metoclopramide; nitrendipine; murine leukemias.ine leukemias.
HIV Proteins-Mediated Alterations in T Cell Interactions with the Extracellular Matrix Proteins and Endothelium (Short Communication).
TOMASZ MROWIEC, CEZARY MELCHAR and ANDRZEJ GÓRSKI

Abstract. Recent data point to the ímmunoregulatory role of extraceIIuIar matríx (ECILf) proteíns ín the process of T cell activation. We studied the effects of HIV proteins on those interactions as well as on endothelial adhesion molecule expression. Gp 120 and gp 41 inhibited CD3-triggered T cell co-stimulation by the ECM proteins, while Nef was stimulatory. T cell adhesion to the ECM proteins was affected in a similar fashion. Furthermore, tat induced E-selectin (but not VCAM-1 and ICAM-1) endothelial expression. Those findings may be relevant for the understanding of immunopathology of AIDS.

Keywords: HIV proteins; endothelium; gp 41; gp 120; Nef; T cells. cells.

Vol. 45, No. 4, 1997

Mini-Review

TNF Receptors – How They Function and Interact.
JACEK BIGDA and HELMUT HOLTMANN

Abstract. Tumor necrosis factor (TNF) is a pleiotropic cytokine regulating various immune and inflammatory reactions. It induces cellular responses upon binding to specific cell surface receptors of two distinct molecular species – p55 and p75 TNF receptors (TNF-Rs). Cloning of the respective cDNAs and obtaining the recep- tor-specific molecular tools such a agonistic and blocking monoclonal antibodies, TNF muteins with exclusive ability to bind to only one receptor type, as well as generation of TNF-R mutants by site-directed mutagenesis enabled researchers to answer key questions of the biology of initial steps in the cascade of TNF signal transduction. The presented mini-review describes the mode of function of the two receptors and discusses the possible ways of interaction between them.

Keywords: tumor necrosis factor (TNF); TNF receptors; signal transduction; cytokines; inflammation.mation.
Integrins and Bone – Cell Adhesion and Beyond
WOJCIECH .J. GRZESIK

Abstract. Integrins are a superfamily of cell surface receptors involved in cell-cell and cell-matrix adhesion. Integrin-mediated interactions are involved in the regulation of numerous cellular functions such as fer- tilization, implantation, cell differentiation and migration during embryonic development, maintaining tissue architecture, blood clot formation and retraction programmed cell death, tumor growth and metastasis formation, lymphocyte homing and response of cells to mechanical stresses. This broad spectrum of activity is achieved by combining the ability to create mechanically functional junctions (cell-matrix and cell-cell) and signal-transducing capabilities. Osteoblasts and osteoclasts express speciftc integrin receptors and the pattern of expression varies depending on the stage of cell differentiation. Interactions of integrins with bone-matrix adhesive proteins are thought to be important for regulating the tissue integrity and may provide a local, responsive regulatory system of osteoblastic differentiation as well. Osteoclasts most likely attach to osteopon- tin exposed on the bone surface via the classic vitronectin receptor a ß3 and this binding may be crucial to their bone resorption activity.

Keywords: integrins; bone matrix; osteoblasts; osteoclasts; arginine-glycine-aspartate.artate.

Clinical Immunology

Immunological Status of Septic and Trauma Patients. II. Proliferative Response and Production of Interleukin 6 and Tumor Necrosis Factor a by Peripheral Blood Mononuclear Cells from Septic Survivor, Nonsurvivor and Trauma Patients: a Correlation with the Survival Rate.
BARBARA ADAMIK, MICHAL ZIMECKI, ANDRZEJ WLASZCZYK and ANDRZEJ KÜBLER

Abstract. We have studied proliferative response to phytohemagglutinin (PHA) and bacterial lipopolysac- charide (LPS) of peripheral blood mononuclear cells (PBMC) and their ability to produce IL-6 and TNF-a in vitro in: septic survivors, nonsurvivors and with multiple injury patients. Blood samples for determination of PBMC reactivity were taken upon admission and after 1, 2 and 5 days later. The proliferative response of lymphocytes, both spontaneous as well as PHA- and LPS-induced, was generally higher in septic nonsurvivor patients comparing to other groups. Markedly elevated LPS-induced proliferation (particularly on day 3) was a bad prognostic sign for survival. The ability of PBMC from the patients to produce IL-6 in culture was depressed in nonsurvivor septic patients and elevated in trauma patients, comparing to control donors. More profound differences were found with respect to TNF-a production which was deeply depressed (both spon- taneous and induced) in septic nonsurvivors. In contrast, TNF-a production in septic survivors was significan- tly higher with a peak response on day 3. Trauma patients, on the other hand, had significantly increased ability to produce TNF-a on day 1 and 2, declining thereafter. The data presented in this report reveal that low production (or even anergy) with respect to synthesis of proinflammatory cytokines in uitro, as well as increased spontaneous proliferation of PBMC and proliferation in response to LPS, are significantly cor- related with the fatal outcome of the septic shock.

Keywords: sepsis; trauma; proliferative response; peripheral blood mononuclear cells; intereleukin 6; tumor necrosis factor a.ctor a.
Does Cyclophosphamide Combined with Methylprednisolone Affect the Expression of Leukocyte Function Associated Antigen 1 in Refractory Rheumatoid Arthritis(Short Communication).
JAN K. ŁĄCKI WOLFGANG MLTLLER-RUCHHOLTZ and STEFAN H. MACKIEWICZ

Abstract. In the present study, in order to get a better insight into the mechanismof action of cyclophos- phamide (CY) in rheumatoid arthritis (RA), we monitored the changes in lymphocytes eypression of leukocyte function associated antigen 1 (LFA-1). A group of 28 patients with refractory severe RA were treated with CY and methylprednisolone (MO) intravenously. Using llow cytometry we evaluated the changes in LFA-1 molecule expression on peripheral lymphocytes. In the analyzed group of patients the proportion of LFA-1 „dim” cells was reduced. After the treatment the ratio was partly normalized. Twelve months after cessation of the therapy high proportion of LFA-1 „dim” was observed only among CY/MP treated patients. The changes were related to clinical improvement. Based on the obtained data, it seems, that the treatment affecting the expression of LFA-1 may slow down lymphocyte migration and by that limit chronic inflammation within the synovium.

Keywords: rheumatoid arthritis; cyclophosphamide; methylprednisolone; leukocyte function association an- tigen 1.n an- tigen 1.
CD3-/HLA-DR+, CD3+/HLA-DR+ Cell Levls and PHA Stimulation of Lymphocytes e Obtained from Women with Recurrent Spon aneous Abortions and Subjected to Partner’s Lymphocyte Immunization.
JACEK WILCZYŃSKI, ZOFIA SUŁOWSKA, ANDRZEJ MALINOWSKI, KRZYSZTOF ZEMAN, MARIAN SZPAKOWSKI, HENRYK TCHÓRZEWSKI and LECH PODCIECHOWSKI

Abstract. It is assumed that 80% of recurrent spontaneous abortions (RSA) of unknown etiology are of immunological origin. The proposed experimental treatment of those patients could be immunization with paternal lymphocytes, though the exact mechanism throug zá on alters the rnumber andltactifvity of parl unknown. Some authors suggest that lymphocyte alloimmuni ticular subpopulations of peripheral blood lymphocytes. The aim of this study was to evaluate the proliferative O activity of lymphocytes stimulated with phytohemagglutinin (PHA) and the percentage of peripheral lym- hoc tes carrying CD3+/HLA-DR+ and CD3-/HLA-DR+ markers, present in 32 selected for this study RSA women subjected to paternal lymphocytes’ alloimmunization performed prior to conception and in early pregnancy. We conclude that, the post-immunization changes seem to have different quantitative and qualita- tive character comparing to those seen in normal pregnancy. They seem to avert unfavourable immunological phenomena observed in RSA women, allowing for successful continuation of pregnancy.

Keywords: recurrent spontaneous abortions; paternal lymphocytes immunization; phytohemagglutinin; lymphocyte subpopulations.ubpopulations.
Polymorphism of the Fourth Component of Complement (C4) and Three Clinical Pictures of Hepatitis B Virus Infection in Children
IRMA KACPRZAK-BERGMAN and JOLANTA HAŁASA

Abstract. The fourth component of complement (C4) phenotype and allele frequencies were studied in 3 groups of unrelated children affected with hepatitis B virus (HBV) infection (25 with glomerulonephritis (GN), 39 with chronic active hepatitis (CAH), 24 with Gianotti-Crosti syndrome (GS-S)) and in 100 unrelated healthy individuals (H). Ten phenotypes in GN, 14 in CAH, 10 in GC-S and 23 in H were found. Some rare phenotypes appeared in the individual cases of ill children. Statistical analysis was performed by chi-square test. Each group was compared to the others. We found significantly higher frequency of the A3B2,Q0 phenotype in CAH than in H as well as significant increase of the A*4 gene frequency in GC-S as compared to H. However, these events could be caused by limited number of tested patients. We suspect that the as- sociation between C4 alleles and immune-complex diseases, studied here, does not rather exist.

Keywords: fourth component of complement alleles; hepatitis B virus diseases; children.ildren.
Cellulsr Immunology Ultrastructure and Functional State of Rabbit Lymphoid Cells after Repeated Exposure to Lipopolysaccharide and External Heating.
PETR P. MURZENOK, NINA I. NETUKOVA and TATJANA I. ZHYTKEVITCH

Abstract. The ultrastructure and proliferative activity of lymphoid tissue cells in rabbits after repeated ex- posures to lipopolysaccharide (LPS) or external heating were studied. It was shown that in conditions of the same increase in body temperature the production of IL-1-like activity by blood and splenic cells was inhibited to the same extent in both groups of animals. It was suggested that one of the causes of LPS-induced tolerance may be events provoked by temperature factor per se. There were similar changes in activity of epithelial cells in the thymus, reticular cells in the spleen, in activation of mast cells in these organs and in proliferative processes in the thymus after both exposures, however inhibition of proliferative activity of splenic and lymph node cells was observed only in LPS-treated animals.

Keywords: ultrastructure; proliferative activity; lymphoid tissue cells; lipopolysaccharide-induced tolerance; external heating.eating.
Loss of T Cell Receptor Diminished Tumorige nicity of Thymocyte-Derived Lymphoma Cells in the T Cell Receptor.
MARCIN KOBZDEJ, .JANUSZ MATUSZYK, EWA ZIOŁO and LEON STRZĄDAŁA

Abstract. Mice with transgenic T cell receptor (TCR) recognizing H-Y male antigen developed spontaneous lymphomas originated from immature thymocytes, with the surface expression of transgenic TCR and CD4/CD8 co-receptors. During in vitro long-term culture (3 months) some lymphoma cell lines lost the surface expression of TCR and co-receptors. Interestingly, the proteins of transgenic receptor were expressed intracel- lularly but TCR was not detectable on the surface of the in vitro selected subline in contrast to TCR-positive parental cells maintained in vivo. TCR-negative subline has been found to be slowly growing in vivo (i.p. injection) and less tumorigenic (s.c. injection) than parental TCR positive lymphoma. It seems that the in vivo interactions of lymphoma cells with microenvironment preserve their TCR expression and endow with growth advantage, while the selected in vitro TCR-negative cells lose the tumorigenic potential.

Keywords: T cell receptor (TCR) transgenic mice; tumorigenicity; TCR anti-H-Y. TCR anti-H-Y.
Effect of Lactoferrin on Proliferation and Dif ferentiation of the Jurkat Human Lymphoblastic T Cell Line.
BAO YUAN BI, ANNE-MARIE LEFEBVRE, DANUTA DUŚ, GENEVIČVE SPIK and JOËL MAZURIER

Abstract. The effect of human lactoferrin on the human lymphoblastic T cell line (Jurkat) was tested with regard to proliferation and differentiation. Lactoferrin enhanced cell proliferation in a serum-reduced (1 % fetal calf serum) culture. The stimulatory effect of lactoferrin on proliferation depended on the degree of iron saturation but the amplitude of the effect was low, similar to that obtained in the presence of serum transferrin. The proliferation stimulatory effect of lactoferrin was not observed in the presence of 10% fetal calf serum (FCS) in the culture medium. These results suggest that Fe-lactoferrin can substitute for Fe-transferrin during the prolonged culture of cells in a low serum concentration. Iron-saturated lactoferrin was also shown to promote T cell differentiation. Jurkat cells, when exposed to iron-saturated lactoferrin in the presence of 10% FCS, gradually exhibited a decrease in the cell volume, cell surface density of CD71 antigen, the nuclear incorporation of [methyl-3H]thymidine, but an increase of the percentage of cell population in the GO/1 phase of the cell cycle. These modifěcations were accompanied by the appearance of CD4 antigen at the cell surface. Therefore, in the continuous presence of lactoferrin, proliferating cells slowly enter into quiescence state, undergoing cell differentiation.

Keywords: lactoferrin; T lymphoblasic cells; proliferation; differentiation.iation.
Morphine Modulation of Thioglycollate-Elicited Peritoneal Inflammation in the Goldfish,Carassius auratus.
MAGDALENA CHADZIŃSKA, SZCZEPAN JÓZEFOWSKI, JANUSZ BIGAJ and BARBARA PŁYTYCZ

Abstract. Intraperitoneal (i.p.) injection of adult goldfish with 3% thioglycollate (TG) induces an acute peritone- al inflammation connected with a massive influx of inflammatory leukocytes mainly of the head kidney origin. The number of peritoneal exudate cells retrieved on day 2 of the inllammatory response is significantly lower in fish injected with TG and morphine (20 mg/kg b.w.) than in animals injected with TG only. The morphine effect was totally antagonized in ftsh injected 20 min earlier with naltrexone (1 mg/kg), a well-known blocker of opioid receptors. Light microscopy of the head kidney Epon sections revealed that basophilic granulocytes are common in the control PBS-injected fish and even more frequent in flsh injected with morphine only. In sharp contrast basophils are very rare in the head kidneys from animals with the TG-induced peritoneal inflamma- tion, while they are more numerous in fish injected with TG and morphine. Supposedly, the basophilic granulocytes might be involved in the inhibitory effects of morphine on the acute inflamation in goldfish. An involvement of the head kidney in the morphine modulation of the inflammatory response is strongly suppor- ted by the detection of opioid receptors in the head kidney cells suspension.

Keywords: goldfish; peritoneal inflammation; head kidney; morphine; naltrexone; naloxone; opioid receptors.oid receptors.
Investigations on Human Lutropin from Pituitary Gland and Urine.
EWA KUROWSKA, IWONA E. KOCHANOWSKA, JANUSZ BORATYŃSKI and APOLINARY SZEWCZUK

Abstract. Two newly isolated monoclonal anti-ßhLH antibodies did not recognize both human chorionic gonadotropin (hCG) and three synthetic peptide fragments corresponding to ßhLH sequence. These antibo- dies were applied for ELISA of human luteinizing hormone (hLH), however, sensitivities of these assays were much lower than those with two other monoclonal antibodies obtained ealier in our laboratory. No significant differences between assays of hLH from pituitary and urine with 6 different pairs of monoclonal antibodies (4 anti-ß and 1 anti-a-subunit) were noticed. The highest hLH concentration in urine samples collected during 53 menstrual cycles of 6 women was demonstrated at different times of the day. In 14 out of 52 „preovulatory urine” portions, preserved for several weeks, over 80″/o increase of assayed hLH was shown. The highest assays of hLH from pituitary and urine were noted at pH 7.5-9.0. Using affinity chromatography the whole pituitary hLH was bound to ConA-Sepharose column and was eluted with a-methyl-D-mannoside as a single peak. However, a small part of hLH from urine was eluted from the column as non retarded peak. When HPLC analysis with DEAE column was performed, pituitary hLH was separated in two fractions while lutropin from urine was eluted as a single peak.

Keywords: human lutropin; human luteinizing hormone (hLH); pituitary hLH; hLH in urine. urine.
Further Investigations on Thymopentin-Like Fragments of HLA-DQ and Their Analogs.
ZBIGNIEW SZEWCZUK, ZBIGNIEW WIECZOREK, PIOTR STEFANOWICZ, ANDRZEJ WILCZYŃSKI and IGNACY Z. SIEMION

Abstract. Recently we showed that the fragments of HLA-DQ with the Thr-Pro-Gln-Arg-Gly-Asp- -Val-Tyr-Thr and Gln-Arg-Gly-Asp-Val-Tyr-Thr sequences strongly suppress the immune response, while their shorter analogs, Arg-Gly-Asp-Val, Arg-Gly-Asp-Val-Tyr and Gln-Arg-Gly-Asp-Val-Tyr, show very weak stimulatory activity with respect to humoral immunological response. The fragments contain the sequen- ce. which is very similar to thymopentin (pentapeptide Arg-Lys-Asp-Val-Tyr, an active fragment (32 – 36) of thymopoietin, an immune , system activator produced in thymi), and at the same time contains the Arg-Gly-Asp (RGD) sequence, known as an inhibitor of adhesion processes. In the present study we found that a hexapeptide: Arg-Gly-Asp-Val-Tyr-Thr is the smallest size fragment of HLA-DQ having both cellular and humoral immunosuppressive activity. We also found that linear and cyclic fragments of HLA-DQ do not affect cell line production of various cytokines, what suggests that the mechanism of interactions of these peptides with the immunological system is different as compared with most other known immunosuppressors.

Keywords: peptide immunosuppressors; fragments of HLA-DQ; interleukins.eukins.
Conjugation of the Monoclonal Antibody 17-lA with the Nitroacridine Compound C921 with the Poly-L-lysine as an Intermediate Agent.
MARIA PAPROCKA, JANUSZ BORATYŃSKl, DANUTA DUS, HALINA KUŚNIERCZYK and CZESŁ.AW RADZIKOWSKI

Abstract. Monoclonal antibody 17-lA specific for human gastric carcinoma was coupled directly or indirectly, using poly-L-lysine as an intermediate, with nitroacridine compound C921. The aim of the study was to obtain selectively cytotoxic immunoconjugates for experimental therapy. Directly coupled conjugates retained an- tibody specificity towards cells of several human adenocarcinoma lines but were non cytotoxic in uitro, whereas conjugates obtained with the use of intermediate poly-L-lysine expressed only low, non-specific cytotoxicity, probably exerted by the poly-L-lysine content.

Keywords: 17-lA monoclonal antibody conjugate; poly-L-lysine; 1-nitroacridine; human colon adenocar- cinoma.nocar- cinoma.

Vol. 45, No. 5-6, 1997

Review

Classification of Cytokines According to the Receptor Code.
ANNA D.INGLOT

Abstract. 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. CHAO

Abstract. . 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. MACDERMOT

Abstract. .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. MACDERMOT

Abstract. . 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. KAHAN

Abstract. . 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 WIĘDŁOCHA

Abstract. 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 JAMES

Abstract. 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 WIECZOREK

Abstract. 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 WIECZOREK

Abstract. 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 SIDORCZYK

Abstract. 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 RZESKI

Abstract. 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 PIETRUSKA

Abstract. 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-STAUBACH

Abstract. . 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 WIKTOROWICZ

Abstract. 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. LIBERSKI

Abstract. 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. SHUKANOVA

Abstract. 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.

Vol. 45, 1997

Vol. 45, No. 1, 1997

CONTENTS

Reviews

Cellular Immunology

Experimental and Clinical Therapy

Various

Vol. 45, No. 2-3, 1997

CONTENTS

Review

Clinical Immunology

Immunochemistry

Various

Vol. 45, No. 4, 1997

CONTENTS

Mini-Review

Clinical Immunology

Vol. 45, No. 5-6, 1997

CONTENTS

Review

Immunochemistry

Experimental Oncology

Various