Vol. 44, 1996

Vol. 44, No. 1, 1996

CONTENTS

Mini-Review

  • Cytokines in the Treatment of Hematological Disorders: Recent Progress and Perspectives
    TADEUSZ ROBAK

    Abstract. Cytokines are involved in hematopoiesis by regulating proliferation, differentiation and cellular functions of various lineages of hematopoietic cells. There is an increasing range of clinical conditions in which cytokines are involved as therapeutic agents. One of the most advanced and successful applications is the stimulation of hematopoiesis by the colony stimulating factors (GM-CSF and G-CSF) and erythropoietin. Hematopoietic growth factors are effective in accelerating recovery from neutropenia after chemotherapy and bone marrow transplantation and in reducing incidence of infections. Interferon a (IFN-a ) proved a useful therapeutic agent for chronic myelogenous and hairy cell leukemias as well as for multiple myeloma and non-Hodgkin’s lymphoma. Interleukin 2 is the only cytokine apart from IFN-a accepted as antineoplastic agent. It may be useful as adjuvant therapy in the hematological malignancies. It may be supposed that in the near future new recombinant cytokines will be introduced in the treatment of blood diseases.

    Keywords: interferon a , interferon g ; hematopoietic growth factors; blood diseases; clinical applications of cytokines.

  • Natural Human Interferon a May Act Differently When Given Parenterally or Orally to Patients Chronically Infected with Hepatitis B Virus
    JERZY A. GEORGIADES

    Abstract. A literature review of patients chronically infected with hepatitis B virus (HBV) treated with natural human interferon a (nHuIFN-a ) given parenterally every day for 28 days revelated that even the daily dose of (5 x 106 IU) of IFN-a inhibits cellular metabolism. As a result of metabolic block, the number of blood elements were diminished. Furthermore treated patients recorded several different adverse reactions. In cont- rast, among patients treated with the oral form of nHuIFN-a no metabolic block occurred and no adverse reactions were seen, even though the therapy lasted much longer. Two years after initiation of parenteral IFN-a therapy, the loss of HBV-BeAg was 53.8% and 28.8% of the patients had undergone seroconversion. In contrast, 77% of patients on oral interferon lost HBV-BeAg and 74% serconverted and normalized their biochemical liver function. The results suggest that the nHuIFN-a given orally and parenterally activate two different mechanisms responsible for virus elimination.

    Keywords: interferon alpha (IFN-a ); cytokines; mode of action; chronic active hepatitis; oral form of IFN-a ; treatment of hepatitis B.

Immunochemistry

  • Effect of a Proline-Rich Polypeptide on IgG Level in Chickens
    MARIA JANUSZ and BARBARA BŁASZCZYK

    Abstract. We have previously shown that a proline-rich polypeptide (PRP) from ovine colostrum expresses an immunoregulatory activity in mice on humoral and cellular immune response. The results presented in this communication show that PRP also modulates the immune response in chickens. PRP administered to the chickens in portions before, simultaneously or after the antigen, stimulated the antibody production. The enhancement of the immune response was also observed when a single dose was injected to the chicken before or simultaneously with the antigen. On the other hand, the single doses of PRP applied to the chickens after the immunization caused an inhibition of the production of antibodies.

    Keywords: proline-rich polypeptide; IgG antibodies.bodies.

  • Interaction of Human Erythrocyte Glycophorin with Immunoglobulins G
    JOANNA MIKULSKA, WOJCIECH GORCZYCA and JÓZEF LISOWSKI

    Abstract. We have found that human glycophorin and asialoglycophorin interacted with human non-immune IgG. To characterize quantitatively the interaction between glycophorin and IgG we elaborated a direct solid-phase radioimmunoassay. We showed that the binding of IgG was reversible and saturable within the range of IgG concentrations used. Glycophorin bound higher amounts of aggregated than non-aggrega- ted immunoglobulins. The apparent association constant for non-aggregated human IgG was 5.45 ± 0.93 x 105 M-1 and 1.13 ± 0.78 x 106 IgG molecules were bound per 1 picogram of the glycophorin. The binding of glycophorin occurs within the F (ab’)2 fragment of IgG, mainly.

    Keywords: erythrocyte; glycophorin; receptors; ligand-receptor interactions; immunoglobulin G.ulin G.

  • Antigenicity of Human and Horse Myoglobins
    IWONA E. KOCHANOWSKA, MARIANNA KUROPATWA, ANDRZEJ RAPAK and APOLINARY SZEWCZUK

    Abstract. Rabbit antisera against human myoglobin and horse myoglobin cross-reacted with both myoglobins but only one of them recognized human hemoglobin. Two mouse monoclonal antibodies anti-human myo- globin were obtained, but only one of them (No. 49) cross-reacted with horse myoglobin. Antibody No. 49 and rabbit antibodies reacted also with apo-, FITC- and treated with hydrochloric acid or TPCK-trypsin horse myoglobin, but their binding to myoglobin pretreated with NaOH was reduced. Thirteen peptides overlapping sequence of human myoglobin were synthesized on polyethylene pins. Rabbit and mouse polyclonal an- tibodies reacted with some of these peptides but no reaction was noted with mouse monoclonal antibodies. Two monoclonal antibodies were applied for specific immunoassay of human myoglobin.

    Keywords: antigenicity, human myoglobin, horse myoglobin, immunoassay of myoglobin, polyclonal and monoclonal antibodies.bodies.

  • Inhibition of Mouse Liver Cytochrome P-450 by Gram-Negative Bacteria Lipopolysaccharides
    WIESŁAW KACA, MILAN MARA and JANA OCENASKOVA

    Abstract. The ability of bacterial endotoxins, of different origin, to modify the level of mouse liver cytochrome P-450 was investigated. Endotoxins, (lipopolysaccharides, LPSs) were isolated from Proteus, Escherichia, Salmonella, Bacteroides and Coxiella strains. The most potent inhibitor of cytochrome P-450 activity was S. typhi 0101 LPS, which at a dose of lm g/mouse reduced the cytochrome P-450 activity to 59%. E. coli 055: B5, S. typhimurium, P. mirabilis 03, and C. burnetii LPSs, at dose 10 m g/mouse, decrease cytochrome P-450 level from 56 to 69%. B. ouatus LPS significantly suppressed the expression of cytochrome P-450 only at the highest dose used – 100 m g/mouse. The comparison of inhibitory activity of P. mirabilis complete, S and R types of LPSs indicate that lipid A portion of LPSs are sufficient to decrease the cytochrome P-450 level. However, the core oligosaccharide of LPS significantly enhance that inhibition. The isolated O-specific polysaccharide part of P. mirabilis 03 LPS did not decrease cytochrome P-450 level. The comparison of biological activity of Proteus LPSs, tested by chromogenic Limulus amebocyte lysate (LAL) test, demonstrated the enhancement effect of O-polysaccharide part of tested LPSs.

    Keywords: Proteus lipopolysaccharides; mouse cytochrome P-450; non-enterobacterial endotoxin.

  • Haptoglobin Concentration in Serum and Other Body Fluids Measured by Comparison of Its Reactivity with Hemoglobin and Concanavalin A
    IWONA KĄTNIK and JOANNA JADACH

    Abstract. Haptoglobin (Hp) concentration in normal human sera was determined either by means of the reaction with hemoglobin (Hb) or by reaction with the lectin concanavalin A (Con A). The ratio of H -Con A 1.02±0.68 g/1) to Hp-Hb (1.10±0.58 g/1) equal 0.96±0.32 was found to be characteristic for sera from healthy subjects. Measurements were also carried out for some pathological body fluids. Values of the ratio Hp-Con A/Hp-Hb in ascitic fluid, pleural effusion and cerebrospinal fluid were 1.72± 1.13 0.42±023 and 0.15±0.18, respectively. Haptoglobin present in urine and saliva was not recognized by Con A, whereas could form a complex with hemoglobin. Hp concentration determined by this method reached 2 mg/l.

    Keywords: haptoglobin; concanavalin; glycosylation.lation.

Cellular Immunology

  • Lactoferrin Inhibits Proliferative Response and Cytokine Production of TH1 but Not TH2 Cell Lines
    MICHAŁ ZIMECKI, JOEL MAZURIER, GENEVIEVE SPIK and, JUDITH A. KAPP

    Abstract. Bovine (BLF) and human (HLF) lactoferrin inhibit proliferation and cytokine production by an antigen specific TH1 cell line stimulated with antigen presenting cells (APC) and antigen. Beside the inhibitory effects on TH 1 cells we observed a decrease of interleukin 2 receptor (IL-2R) expression on these cells contacted with LF. Lactoferrin (LF) did not inhibit proliferation of indicator HT-2 cells to IL-2 suggesting lack of interference with IL-2/IL-2R interaction by LF. No inhibitory effect was observed on proliferation and cytokine production by TH2 cell lines and no changes in interleukin 4 receptor (IL-4R) levels were found with regard to TH2 cell line. We conclude that LF has differential activity with regard to effector functions of antigen-specific T cells by inhibiting TH 1 but not TH2-mediated immune responses.

    Keywords: lactoferrin; THl, TH2 cells.Hl, TH2 cells.

  • Effect of Exogenous Tumor Necrosis Factor a, Interleukin 6 and Interferons on Vesicular Stomatitis Virus Replication in Human Placenta and Amniotic Membrane Organ Cultures
    EDYTA PARADOWSKA, ZOFIA BŁACH-OLSZEWSKA, MARZENA GIERLIŃSKA and JANUSZ WOYTOŃ

Abstract. Effects of exogenous cytokines on replication of vesicular stomatitis virus (VSV) in amniotic mem- brane and placental organ cultures (OC) were studied. We compared the effects observed in OC and establi- shed human carcinoma cell lines: A549 and HEp-2. Recombinant human tumor necrosis factor alpha (rHuTNF-a ), added to amniotic membrane, villous, or decidual OC at concentrations of 30 to 3000 U/ml, potentiated VSV replication by 10-1000 fold. Addition of 5 to 10000 U/ml of recombinant human inter- leukin 6 (rHuIL-6) to OC from 5 placentas was without effect on VSV growth, except one culture in which enhanced VSV replication has been observed. rHuTNF-a was found to have no effect on VSV growth in HEp-2 and A549 cell cultures. In contrast, the placental OC were sensitive to antiviral activity of natural interferons (IFNs): a , ß and recombinant IFN-g , although A549 cells were 5 to 10 fold more responsive to the cytokines.

Keywords: cytokine; placenta; virus replication.

Various

  • Serological Confirmation of Human Herpesvirus Type 6 Infection in Different Age Groups
    AGNIESZKA BIESIADECKA, BOGUMIŁA LITWIŃSKA and MIROSŁAW KAŃTOCH

    Abstract. Human herpesvirus type 6 (HHV-6) infection was proved by IgG immunofluorescence method. 354 sera at various age groups were tested and 161 sera from children in more detail. There was no significant differences in our results and obtained by other authors, when testing sera from adults. Our investigation of younger children sera confirmed that mother antibodies are transferred, but also suggest the possibility of virus vertical infection. Our study also confirmed that antibodies are acquired very early in life in the same age group as exanthema subitum does.

    Keywords: human herpesvirus type 6 (HHV-6); age prevalence of IgG HHV-6; mother antibodies; vertical virus transmission.ission.

  • Seleno-Organic Compounds as Immunostimulants: an Approach to the Structure-Activity Relationship
    ANNA D. INGLOT, JACEK MŁOCHOWSKI, JANINA ZIELIŃSKA-JENCZYLIK, EGBERT PIASECKI, TOMASZ K. LEDWOŃ and KRYSTIAN KLOC

    Abstract. Our studies on the seleno-organic compounds were focused at their activities as the modest cytokine inducers in human peripheral blood leukocyte cultures. Our bioassays used in the screening methods were based on the quantitative determinations of mainly two types of cytokines: interferons (IFNs) and tumor necrosis factors (TNFs). More recently we have found that several of the compounds have direct immuno- tropic actions in vitro and in vivo, in mice and in chickens. The paper summarizes the data related to the cytokine-inducing activity of 65 seleno-organic compounds divided into 4 groups according to their chemical structures. The reference compound was ebselen, the well known experimental drug with various biological ac- tivities. Approximately 50% of the compounds were found to be active in our bioassays. The selected com- pounds induced also IL-6 and GM-CSF. Their activities were clearly correlated with defined chemical structures as well as with the presence of selenium. We suggest that some of the selected by us compounds, other than eb- selen, are interesting as immunostimulants and potential antiviral agents and cytokine inducers active in humans.

    Keywords: interferon and tumor necrosis factor inducers; cytokines; immunostimulants; human peripheral blood leukocytes; seleno-organic compounds; ebselen.bselen.

  • Spontaneous Motility and Chemotaxis of Neutrophils Is Influenced by Glycocorticosteroid Therapy
    TERESA ŻAK-NEJMARK, RENATA JANKOWSKA, JÓZEF MAŁOLEPSZY, MAREK JUTEL, MARIA KRAUS-FILARSKA and GRAŻYNA NADOBNA

Abstract. The migration of neutrophils is an important part of the allergic inflammatory response. The aim of our study was to investigate the effect of glucocorticosteroids (GCS) on the stimulated and unstimulated migration of neutrophils. The study comprised 103 asthmatics including 44 subjects under GCS therapy (20 GCS resistant and 24 GCS sensitive) as well as 96 healthy control individuals. Unstimulated (random) motility as well as chemotactic response towards fMLP (l0-8 mol/1) were determined after Boyden method. Neutrophil motility was determined by the distance of the leading front in filter. In both resistant and sensitive asthmatics under GCS therapy we observed significantly increased unstimulated motility as compared to normal controls (p < 0.001). However, this effect was not demonstrated in the GCS untreated group. Neutrophil chemotaxis towards fMLP was increased in GCS untreated group as compared to healthy controls (p < 0.05). In GCS sensitive subjects the chemotatic activity was decreased. In GCS resistant asthmatics it was moderately increased. We conclude that increased unstimulated neutrophil motility might be one of the immunosuppressive mechanisms of GCS by preventing the cell accumulation at the sites of inflam- mation.

Keywords: asthma; glucocorticosteroid therapy; cell motility.cell motility.

Vol. 44, No. 2-3, 1996

CONTENTS

Cellular Immunology

  • Effect of Interferon a on HLA-DR Expression by Human Buccal Epithelial Cells
    J. KELLY SMITH, DAVID S. CHI, GUHA KRISHNASWAMY, SUJATA SRIKANTH, SCOTT REYNOLDS and STEVEN L. BERK

    Abstract. We have studied the effect of interferon a (IFN-a) on MHC class II expression by human buccal epithelial cells (BEC), and mRNA expression by BEC and mucosal-associated mononuclear cells (MAMC). In 6 experiments, freshly collected BEC were suspended at a concentration of 1.0 x 105/ml in RPMI 1640 and incubated in the presence of 0-10000 IU/ml of human lymphoblastoid IFN-a (HuIFN-a ). Zero and six hour samples were analyzed by single color flow cytometry using FITC-labeled murine IgG1 monoclonal antibody to HLA-DR. Preparations were also analyzed for expression of cytokine transcripts (IL-2, IL-4, IL-5, IL-6, IL-8, IFN-g , GM-CSF) by reverse transcriptase polymerase chain reaction (RT-PCR). Increasing concen- trations of IFN-a resulted in proportionate increases in the percentage of HLA-DR+BEC (r = 0.7897, p = 0.0627) and in the percentage of HLA-DR+staining at higher intensities (101 to 10z log fluorescence intensity) (LFI) (r = 0.4010, p = 0.0424). The percentage of HLA-DR + BEC rose from a mean of 1.5 % with no IFN-a to 7 % with 10 000 IU/ml IFN-a (p < 0.05). The percentage of HLA-DR + BEC staining at 101 to 10z LFI rose from a mean of 8.3o/o with no added IFN-a to 19.2% with 10000 IU/ml IFN-a (p < 0.05). Unstimulated BEC constitutively expressed IL-8 and GM-CSF. IFN-a stimulated preparations also expressed IFN-g , possibly due to the presence of MAMC, which comprised 2-9% of the total cell population. These data indicated that HuIFN-a upregulates MHC class II expression by human BEC, possibly by enhancing IFN-g production by MAMC present in the culture preparations.

    Keywords: human lymphoblastoid interferon; human buccal epithelial cells; MHC class II antigens; cytokine mRNA transcripts; mucosal-associated mononuclear cells.

  • Analysis of Tolerance Inducing Mechanism after Application of Oncogen-Transformed Macrophages
    ANA MARIA WAAGA, JAN-MICHAEL OTTE, MACIEJ KRZYMAŃSKI, VOLKER ECKSTEIN and WOLFGANG MÜLLER-RUCHHOLTZ

    Abstract. The purpose of this study was to examine the expression of T cell receptors (TCR) and their Vß subclasses under the influence of the parental cell line P388D1 and its clones mos2 and mos3, using a mouse model. It was shown, that v-mos oncogene-transformed cells of this line (mos2) induced selective immunological unresponsiveness in vitro. Because the induction of tolerance is of a central importance for the organ transplantation, this phenomenon, found in vitro, was also studied in vivo. We found that the in vivo injection of mos2 cells into mice induced a state of selective noncreactivity. To further analyse these effects, we studied whether specific tolerance is the consequence of a decreased number of essential receptors or receptor families. For this purpose C57BL/6 mice were immunized with cells of the parental line P388D1 or mos2 and mos3 clones. Their spleen and thymus cells were examined phenotypically. The most impressive result of this study was a clearly changed amount of T cells receptors in mos2 immunized mice, in which a state of tolerance was induced. In these mice only the expression of CD3 T receptors as well as that of the Vß11 chains was reduced. In spleen of these mice the CD3 expression was decreased, compared to D1 or nonimmunized control animals by 54-58% and compared to mos3 mice by 38-40%. Even though the differences in the thymus were not very prononced, we still saw a decrease in CD3 stained cells selective in mos2 immunized C57B1/6. The expression of Vß11 chains on the surface of spleen cells of mos2 animals was reduced by 33.3%, on the thymocytes even by 50% comparying to that in nonimmunized mice. Whether the reduced expression of T receptor Vß families is due to changes in the genetic material (cDNA), has to be studied.

    Keywords: tolerance induction; oncogene-transformed macrophages; polymerase chain reaction; T cell receptor; P388D1 (D1) transformed with v-mos oncogene; clones mos2 and mos3.mos2 and mos3.

  • Tumor Necrosis Factor and Interferon Production by Peripheral Blood Leukocytes of Patients with Alcoholic Cirrhosis
    JADWIGA DANILUK, MARTYNA KANDEFER-SZERSZEŃ and LILIANA BOROWSKA

    Abstract. Sixteen male and female patients with alcoholic cirrhosis and 13 healthy controls were included in the study. The level of interferon (IFN) and tumor necrosis factor (TNF) activity was examined in the sera and also in cultures in peripheral blood leukocytes (PBL) after in vitro stimulation with the cytokine inducers. In sera of patients with alcoholic cirrhosis higher IFN and TNF levels were detected than in controls. However, after induction with Newcastle disease virus (NDV), phytohemagglutinin M (PHA) and lipopolysaccharide from E. coli (LPS), PBL from cirrhotic patients produced lower IFN levels in comparison to controls. In contrast to depressed ability to produce IFNs, TNF production was higher in PBL of cirrhotic patients induced by PHA and a low dose of LPS (1 m g/ml). NDV induced comparable levels of TNF in both groups. It appears likely that cells of cirrhotic patients were suppressed by an unknown factor or were hyporeactive for IFN production, but synthesis and release of TNF was enhanced, suggesting that cells producing TNF were preactivated in vivo. Mechanism of such preactivation is discussed.

    Keywords: alcoholic cirrhosis; interferon; tumor necrosis factor.

  • Unexpected Reaction of Anti-H-2d Serum with Thymic Lymphoma Cells Derived from Transgenic B6-H-2b TCR anti-HY/Db Mice
    JANUSZ MATUSZYK, EWA ZIOŁO, MARCIN KOBZDEJ and LEON STRZĄDAŁA

    Abstract. Transgenic mice with a ß TCR specific for male antigen (HY) in the context of H-2Db MHC class I were prepared by introduction of transgens into (CS7BL/6J x DBA/2J)F1 hybrid mice (H-2b/d), where only H-2b is selective for maturating thymocytes. Founder transgenic mouse was backcrossed to H-2b MHC background. Serological typing of H-2 antigens revealed that transgenic mice do not express H-2d antigens. Unexpectedly, cells of thymic lymphomas, spontaneously developed in about 45% of old B6 (H-2b) transgenic mice and peripheral blood lymphocytes of about 40% of transgenic mice, reacted with anti-H-2d serum (strain 129 mice (H-2b) immunized with thymocytes and splenocytes of BALB/c mice) but not with monoclonal antibodies against H-2d MHC class I antigens. Anti-H-2d serum has been shown to react with thymocytes but not peripheral blood lymphocytes from non-transgenic H-2b mice. The lymphocytes of transgenic mice reacting with anti-H-2d serum could represent disseminating preneoplastic or neoplastic cells expressing antigen encoded outside MHC region and present on the cell surface of immature thymocytes but not lymphocytes in healthy mice.

    Keywords: TCR anti-HY/Db transgenic mice; thymocytes; thymic lymphomas.

  • Early Supernatants of Mitogen-Stimulated Cells Promote Long-Term IL-2-Dependent of Human T Lymphocytes
    JAN KONRAD SIWICKI and JAN A. STEFFEN

    Abstract. Human wheat germ agglutinin (WGA)- or PHA-activated T lymphocytes from either peripheral blood or normal spleens can be propagated in vitro for several months in the presence of IL-2 but not IL-4 when periodically restimulated with supernatants collected from 24 h cultures of mitogen-stimulated human blood or spleen mononuclear cells. Following several rounds of stimulation with these supernatans some of the spleen-derived T lymphoblast lines acquire the ability of continuous growth (over 100 population doublings) in the presence of IL-2 alone. We postulate that the ability of 24 h supernatants from mitogen stimulated mononuclear cells to maintain T lymphoblast IL-2-dependent growth depends upon yet undefined cytokine(s) released early after activation.

    Keywords: wheat germ agglutinin; T cell; long-term growth.growth.

Experimental Oncology

  • Interferon Production by Peripheral Blood Leukocytes of Patients with Acute Myeloid Leukemia
    MARTYNA KANDEFER-SZERSZEŃ, ANNA DMOSZYŃSKA, LILIANA BOROWSKA, MARIA ADAMCZYK-CIOCH, MAREK HUS, IWONA HUS and JACEK ROLIŃSKI

    Abstract. The ability of peripheral blood leukocytes (PBL) of 13 selected patients with acute myeloid leukemia (AML), and of 10 healthy subjects to produce interferon (IFN) spontaneously and after in vitro induction was tested. Spontaneous IFN production was detected in supernatants of PBL cultures of healthy subjects but was not present in cultures of leukemic cells (leukocytes density 1 x 106 ml). After induction with Newcastle disease virus (NDV) PBL cultures from 5 patients with AML exhibited a low IFN response, while in others IFN titers were similar to controls. The IFN titers induced with lipopolysaccharide (LPS) were lower in leukemic cells than those detected in control cells. The absence or low IFN levels after induction with phytohemagglutinin (PHA) correlated with very small numbers of normal lymphocytes in blood of leukemic patients.

    Keywords: acute myeloid leukemia; interferon production.on production.

  • Serum Soluble Interleukin 2 Receptor a and Soluble CD8 Levels in Patients with Gynecological Undergoing Radiotherapy
    IRENA FRYDECKA, MAŁGORZATA RUSIECKA, KAZIMIERZ KULICZKOWSKI and JAN KORNAFEL

    Abstract. The phenotype peripheral blood lymphocytes, soluble interleukin 2 receptor a (sIL-2Ra) and soluble CD8 (sCD8) were studied in 30 patients with gynecological malignancies before and after ratiotherapy. The absolute count of CD3+, CD4+, CD8+, CDl9+ and CD2S+ lymphocytes before treatment was within normal range. The levels of serum sIL-2Ra and sCD8 were significantly increased before treatment compared to normal controls. After treatment the absolute number of CD3+, CD4+ and CDl9+ lymphocyte subsets significantly decreased. The levels of sIL-2a and sCD8 corrected according to absolute lymphocyte count reached normal range. The decrease of tumor mass correlated with the serum levels of sIL-2Ra and sCD8. Further studies are needed to evaluate the usefulness of these tests in the prognosis and treatment of these diseases. Our study suggests that sIL-2Ra and sCD8 in our patients probably has not been originating from the proteolytic cleavage of membrane bound receptors from peripheral blood mononuclear cells (PBMC). It can be specutaled that sIL-2Ra and sCD8 originated in these patients from activated tumor-infiltrating lymphocytes or may arise from separate alternatively spliced mRNAs coding for soluble vs. membrane forms.

    Keywords: soluble interleukin 2 receptor a; soluble CD8; gynecological malignancies.

  • Impaired CD2S Expression and Soluble CD2S-IL-2Ra Receptor Release by Anti-CD3 Stimulated Peripheral Blood Mononuclear Cells in Hodgkin’s Disease Patients
    IRENA FRYDECKA and GRZEGORZ MAZUR

    Abstract. We examined 22 patients in active phase of Hodgkin’s disease (HD),12 patients in complete clinical remission and 16 healthy subjects for their responsiveness to anti-CD3 stimulation as measured by CD2S antigen expression on peripheral blood mononuclear cells (PBMC) and production of CD2S-IL-2Ra . Simultaneously, the serum levels of soluble interleukin 2 receptor a (sIL-2Ra ) were estimated.0ur studies indicate that PBMC in patients in active phase and in clinical remission have impaired ability to express CD2S antigen after stimulation with anti-CD3 monoclonal antibody. Similarly, the levels of sIL-2Ra in culture supernatants were significantly lower in both groups of patients: in active phase and clinical remission compared with the controls. In contrast, the mean serum level of sIL-2Ra was significantly higher in active phase of the disease compared to that found in patients in clinical remission and controls.0ur studies suggest that sIL-2Ra , derived from PBMC, is not the source of increased levels of sIL-2Ra found in the sera of patients in active phase of HD. Their synthesis and secretion/shedding most probably takes place in the involved tissues.

    Keywords: soluble interleukin 2 receptor a ; anti-CD3; Hodgkin’s disease.

  • Mouse Multivalent IgG(2a, b) Antibody – Ricin A-Chain Immunotoxins Combined with Homologous or Heterologous Interleukin 2 in the Treatment of a Murine Malignant Lymphoproliferation (EL4)
    GABRIELA MOTA, RADU MARCHES, VALENTIN CIALACU, VIVIANA ROMAN, ECATERINA KOZMA, MARINELA MĂRGINEANU, SILVIA DIMA and ION I. MORARU

    Abstract. Two immunotoxins containing ricin A-chain, staphylococcal protein A and mouse anti-Thy 1.2 antibodies of IgG(2a,b) subclass, were prepared. The two multivalent immunotoxins of 750 and 370 kDa and molar ratio A-chain: IgG of 1:2, were used for the treatment of mice bearing ascitic EL4 lymphoma. The immunotoxin-treatment, performed intraperitoneally, was combined or not with homologous or heterologous interleukin 2. The antitumor effects expressed by increase of mice survival time (p < 0.001 as compared with nontreated animals) corresponded to a proportion of 88-90% lymphoma cell-kill by immunotoxin and 93-95a/o by combination treatment (immunotoxin+interleukin 2), as calculated from the relationship between the survival time of nontreated mice and the number of tumor cells inoculated.

    Keywords: immunotoxins; ricin A-chain; interleukin 2; experimental tumors.mental tumors.

Immunopharmacology

  • Antiviral Activity of 2-Chloro-2′ -Deoxyadenosine (2-CdA)
    RENATA ZAHORSKA, NATALIA MAZUR, JANUSZ S. SKIERSKI and PAWEŁ GRIEB

    Abstract. The effect of 2-chloro-2′-deoxyadenosine (cladribine,2-CdA) on replication of herpes simplex type 1(HSV) virus in Vero cells (green african monkey kidney line) was evaluated and compared with that of acyclovir.2-CdA up to 600m g/ml was not cytotoxic. ED50 for 2-CdA was approximately 5 m g/ml and ED 1 oo was approximately 70 tlg/ml, compared with 0.16m g/ml and 1.6 gg/ml for acyclovir, respectively. DNA cell cytometry suggested the progressive S-phase block at concentrations of 2-CdA 0.14m g/ml and higher. The results suggest that 2-CdA is not activated by HSV-encoded kinase, and a weak antiviral effect of the drug results from its interference with DNA replication following activation by kinases of host cells.

    Keywords: 2-chloro-2′-deoxyadenosine; cladribine; herpes simplex virus; cell cycle; DNA content; Vero cells.

  • Induction of Specific Tolerance by 15-Deoxyspergualin (DOS) Treatment after Rat Skin and Kidney Transplantation in Rats. Analysis of Effectivity of Various Protocols of DOS Application
    ANA MARIA WAAGA, MACIEJ KRZYMAŃSKI, KARIN ULRICHS, ANDRZEJ OKO and WOLFGANG MÜLLER-RUCHHOLTZ

    Abstract. The purpose of this study was to analyse the effect of various protocols of 15-deoxyspergualin (DOS) application on skin or kidney graft survival. Following rat strain combinations were used: AS ® LEW (MHC identical/non-MHC-different) and DA ® LEW (MHC-different/non-MHC-different). Reference DOS dose was 2.5 mg/kg, i.p. It was shown that the effect of DOS depended on multiple factors, such as: type of tissue or organ, onset of treatment, drug dose and length of drug application. In skin transplantation graft survival was 32-34 days in AS ® LEW and 24-26 days in DA ® LEW. Kidney graft survived more than 150 days. DOS prolonged skin survival when the application was started earlier than day 8, whereas kidney graft survived only when DOS treatment was started not later than 3-4 days after transplantation. In skin transplantation a dose of 0.3 mg/kg had a small effect – prolongation graft survival up to 4 days. Higher doses induced longer graft survival, however, maximal survival of allogeneic skin was 22 days. In kidney transplantation a dose of 0.3 mg/kg led to prolonged graft survival – up to 150 days. Doses of 2.0-2.5 mg/kg were able to induce specific tolerance. The optimal skin or kidney graft survival was obtained when DOS was applied for 14 days. Shorter than 12-day treatment with DOS led to a shorter graft survival. When donor was pretreated with DOS prolongation of non-allogeneic graft survival was observed. Our results showed that short-term application of DOS is safe and effective. To obtain optimal DOS effect the drug application must be started directly after transplantation.

    Keywords: 15-deoxyspergualin; experimental transplantation; rat kidney transplantation; skin transplantation.

  • In vitro Analysis of the Mode of Action of the Immunosuppressive Drug 15-Deoxyspergualin
    ANA MARIA WAAGA, MACIEJ KRZYMAŃSKI, KARIN ULRICHS, JÖRGEN TREUMER, ANDRZEJ OKO and WOLFGANG MÜLLER-RUCHHOLTZ

    Abstract. The purpose of the study was to analyse the action of the immunosuppressive drug I 5-deoxysper- gualin (DOS) in vitro. We studied: a) the influence of DOS alone and DOS in combination with various monoclonal antibodies on alloantigen stimulation in the mixed lymphocyte culture (MLC), b) the influence of DOS treatment on the MHC class I and II expression of splenocytes, lymph node cells and peritoneal macrophages, c) the influence of DOS treatment on a suppressor cell population. Our study showed that: a) DOS inhibits interleukin 1 (IL-1) secretion by macrophages, leading to reduction of immune response to alloantigens. This effect was neutralized by addition of IL-l; b) DOS treatment has no influence on MHC class II antigen expression, but induces changes of MHC class I expression. After DOS application in a population of spleen macrophages a subpopulation of cells with reduced MHC class I antigen expression appeared. Down-regulation of these molecules was also observed in immunomorphological studies of kidney graft sections of rats treated with DOS after transplantation; c) after DOS treatment suppressor cells were detected in „suppressor” MLC,16-33 days after kidney transplantation. Their activity was confirmed 137 days after treatment with DOS, but were inactive in the case of third party cells. These results suggest that DOS action is based on a blockade of antigen presentation by reducing IL-1 production, down-regulation of MHC class I antigen and by inducing suppressor cell population.

    Keywords: 15-deoxyspergualin; interleukin l; MHC antigens; in vitro studies; mixed lymphocyte culture; monoclonal antibodies; suppressor cells. cells.

  • Monoclonal Antibody Analysis of Glomerular and Renal Interstitial Immune Cells in Steroid-Resistant and Steroid-Responsive Minimal Change Disease in Children
    MAŁGORZATA WĄGROWSKA-DANILEWICZ and MARIAN DANILEWICZ

    Abstract. Glomerular and interstitial leukocyte infiltrates were studied by indirect immunoperoxidaze technique using monoclonal antibodies against all leukocytes, pan-T lymphocytes, helper/inducer CD4 cells, suppressor/cytotoxic CD8 cells, pan-B lymphocytes, monocytes/macrophages and polymorphs in 23 renal biopsy samples obtained from children with minimal change disease (MCD). The cases were classified on the basis of light, electron microscopy and immunofluorescence. Nephrotic syndrome was steroid-sensitive in 15 patients (group I) and steroid-resistant in 8 subjects (group II). Both groups did not differ in the type or number of glomerular immune cells, quantified as immunopositive cells per glomerular cross section. Intraglomerular cellular infiltrates were mainly constituted of monocytes/macrophages; among T lymphocytes T helper/inducer cells predominated over T cytotoxic/suppressor cells. Enumeration of interstitial immune cells showed significantly higher number of all leukocytes and T lymphocytes, quantified as immunopositive cells/mm2, in steroid-resistant group. T helper/inducer cells predominated over T cytotoxic/suppressor cells in the renal interstitium in steroid-responsive and in steroid-resistant MCD. The leukocyte infiltrates did not correlate with a degree of proteinuria at the tine of biopsy and with the renal cortical interstitial volume in both studied groups. Our data suggest, that steroid resistance in minimal change nephropathy in children seems to be related to interstitial T lymphocytes.

    Keywords: minimal change disease; monoclonal antibodies; immunohistochemistry; leukocyte subsets.ocyte subsets.

Various

  • A New Lymphoblastoid Cell Line Defective in Class II HLA Expression
    MARIA MAŃCZAK, JOANNA DUBIS, ANITA KOŚĆ, KATARZYNA BALDY-CHUDZIK, MICHAŁ MACHNICKI, RAFAŁ PACHOLCZYK, MAGDALENA PRUSSAK, BEATA NOWAKOWSKA and PIOTR KUŚNIERCZYK

    Abstract. A lymphoblastoid cell line, HAJ, was derived by in vitro transformation with Epstein-Barr virus of peripheral blood lymphocytes (PBL) from a patient with renal insufficiency awaiting kidney graft. Cell surface expression of class I and class II HLA molecules was determined by flow cytofluorimetry using monoclonal antibodies and compared with that of cell line PAJ similarly derived from a healthy donor. HAJ cells expressed class I antigens at levels comparable with PAJ cells. In contrast, class II antigens were absent from the cell surface of HAJ cells while they were abundant on PAJ cells. Permeabilization and fixation of cells with acetone/formaldehyde solution revealed intracellular Ki-67 antigen but not class II HLA molecules. The genes for HLA-DRß, DQa and DPa were present in the HAJ genome as detected with polymerase chain reaction (PCR) using locus-specific primers amplifying a second exon. In RT (reverse transcriptase) – PCR, transcripts of DQA1 and DPA1 genes were easily detectable in PAJ (positive control) but not in HAJ cells. These results suggest a defect in HAJ cells of transcription of genes for all class II antigens. The cell line HAJ may prove to be an interesting model for in vitro studies of molecular mechanisms of the regulation of class II expression.

    Keywords: HLA class II genes/antigens; gene expression; lymphoblastoid cell line; polymerase chain reaction.

  • Structure of the O-Specific Polysaccharide Chain and Serological Characterization of the Proteus penneri 62 Lipopolysaccharide Compared with the Lipopolysaccharides of the P. penneri Strains
    ZYGMUNT SIDORCZYK, ANNA ŚWIERZKO, KRYSTYNA ZYCH NIKOLAY A. PARAMONOV, EUGENY V. VINOGRADOV, ALEKSANDER S. SHASHKOV and YURIY A. KNIREL

    Abstract. The chemical structure of the O-specific polysaccharide chain of Proteus penneri 62 lipopolysaccharide (LPS) containing N-acetylisomuramic acid was established using acid hydrolysis, solvolysis with anhydrous hydrogen fluoride and 1H and 13C NMR spectroscopy. Cross reactivity of the anti-O-serum P. penneri 62 with a number of other strains of the same species isolated in the USA, Canada, Germany and Poland is discussed.

    Keywords: Proteus penneri; O-polysaccharide; lipopolysaccharide.haride.

  • Natural Human Interferon a Given Orally Has Different Effects on Patients with Distinct Forms of Chronic Viral Hepatitis B
    JERZY A. GEORGIADES, JERZY CABAN, TEOFILA ŻYRKOWSKA-BIEDA, MALGORZATA ZEJC, URSZULA JONAS-SKULINA and ANDRZEJ CIEŚLA

    Abstract. During therapy of chronic viral hepatitis B (CVHB), some patients treated with natural human interferon a (nHuIFN-a ) lozenges failed to respond. These observations triggered studies aimed to determine whether there are markers predicting patients’ response to therapy with nHuIFN-a lozenges. In these studies, 32 patients with CVHB were involved: 20 males and 12 females, 16-61 years of age with proven persistent hepatitis B viremia (HBV). Patients were evaluated for clinical, biochemical liver function, and virological markers of disease. During 300 days of treatment the patients received 75 -150 IU nHuIFN-a daily in form of lozenges. The responders to oral interferon therapy were those who had initially alanine amino transferase (ALAT) level higher than 100 IU (85.7% cure rate) and weak responses were observed among patients who had an initial ALAT level below 100 IU (9.0% response rate). Therefore, ALAT test in patients with CVHB may serve as a predicting indicator of the outcome of IFN lozenges therapy.

    Keywords: natural interferon a ; oral administration; chronic viral hepatitis B; therapeutical efficacy.

  • Influence of Uro-Vaxom on sIgA Level in Urine in Children with Recurrent Urinary Tract Infections
    MIECZYSŁAWA CZERWIONKA-SZAFLARSKA and MAŁGORZATA PAWŁOWSKA

Abstract. We have examined the influence of Uro-Vaxom on secretory IgA (sIgA) level in urine of children with recurrent urinary tract infections. In the group of children treated with antibiotics and Uro-Vaxom, a significant increase of sIgA in the urine was found which persisted for at least 3 months. During this period no infection appeared in 92% examined children.

Keywords: Uro-Vaxom; recurrent urinary tract infection.ection.

Vol. 44, No. 4, 1996

CONTENTS

Mini-Review

  • Retrograde Transport of Acidic Fibroblast Growth Factor as a Part of the Growth Factor Signaling
    ANTONI WIĘDŁOCHA

    Abstract. The plasma membrane represents an impermeable barrier to proteins and other macromolecules. However, certain exogenous proteins are able to cross cellular membranes and gain access to the cytosol. The best examples are bacterial and plant protein toxins, acting on intracellular targets. During last few years the number of known proteins possessing the capability to cross cellular membranes in the reverse direction and reach the nucleus has increased (acidic and basic fibroblast growth factor, interleukin 1, angiogenin, Schwannoma derived growth factor, homeoprotein Antennapedia, HIV-1 Tat protein are some examples). Here, the role of transport of exogenous acidic fibroblast growth factor to the nuclear location as a part of the growth factor signaling is discussed, and the current knowledge on this issue is reviewed.

    Keywords: acidic fibroblast growth factor (aFGF); cellular membranes; transport of aFGF; tyrosine kinase.kinase.

Cytokines

  • Immunomodulatory Activity of Conformationally Restricted Peptides Related to TGF-ß2 90 – 99 Sequence
    ZBIGNIEW WIECZOREK, JACEK J. SŁOŃ and IGNACY Z. SIEMION

    Abstract. Continuing our research on the immunomodulatory activity of the peptides related to the proteins of TGF-ß family, we have synthesized and investigated by Jerne’s plaque forming cell (PFC) test and delayed type hypersensitivity (DTH) test the following linear peptides and their cyclic (disulphide-bridged) analogues : Lys-Thr-Pro-Lys-Ile (Ia) and Cys-Lys-Thr-Pro-Lys-Ile-Cys (Ib), Tyr-Ile-Gly-Lys-Thr-Pro (IIIa) and Cys-Tyr-Ile-Gly-Lys-Thr-Pro-Cys (IIIb), Tyr-Ile-Gly-Lys-Thr-Pro-Lys-Ile (IVa) and Cys-Tyr-Ile-Gly-Lys-Thr-Pro-Lys-Ile-Cys (IVb), Tyr-Tyr-Ile-Gly-Lys-Thr-Pro-Lys-Ile-Glu (Va) and Cys-Tyr-Tyr-Ile-Gly-Lys-Thr-Pro-Lys-Ile-Glu-Cys (Vb). It has been found that the insertion of the sequence of Ia into the disulphide bridged Ib generates a product with enhanced immunosuppressive activity. The similar procedure applied to IIIa, IVa and Va produces cyclic peptides devoided of any activity or of a diminished activity in DTH test. The exchange of Pro residue of Ia D-Pro resulted in the analogue IIa with a preserved immunosuppressive activity. The disulphide-bridged peptide IIb, a cyclic analogue of IIa, exhibited the same activity as IIa in both: PFC and DTH tests.

    Keywords: immunosuppression; immunopeptides; TGF-ß2 immunosuppressive sequences.uences.

  • Colostrinine: a Proline-Rich Polypeptide from Ovine Colostrum Is a Modest Cytokine Inducer in Human Leukocytes
    ANNA D. INGLOT, MARIA JANUSZ and JÓZEF LISOWSKI

    Abstract. A proline-rich polypeptide (PRP), now named colostrinine, molecular weight 18 000, was isolated from ovine colostrum and characterized by Janusz Lisowski et al. The nonapeptide (NP) which is an active fragment of PRP was obtained by chemical synthesis. In mice, PRP has many regulatory effects on the humoral and cellular immune response. The present paper describes PRP as a cytokine inducer. PRP at concentration of 1-100 m g/ml was found to induce production of interferon (IFN) and tumor necrosis factor (TNF) in human peripheral blood leukocytes and in whole blood cultures. The effects were dose related. The identified till now cytokines induced by PRP were IFN-g and TNF-a but many other cytokines may be stimulated also. NP was considerably less active as the cytokine inducer than the natural PRP. Two volunteers given orally once daily for two to three weeks 100 or 200 m g PRP in tablets were found to develop the tolerance to IFN induction and had the modified TNF response. Furthermore, the PRP-treated volunteers showed signs of psycho-stimulation. Taken together our observations suggest that ovine PRP is active in humans and may have therapeutic value as an immunostimulant and/or neurotropic cytokine.

    Keywords: colostrinine; proline-rich polypeptide; interferon g ; tumor necrosis factor; human leukocytes; cytokine inducers; ovine colostrum.

  • Immunological Status of Patients Subjected to Cardiac Surgery: Serum Levels of Interleukin 6 and Tumor Necrosis Factor a and the Ability of Peripheral Blood Mononuclear Cells to Proliferate and Produce these Cytokines in vitro
    ANDRZEJ WŁASZCZYK, BARBARA ADAMIK, GRAŻYNA DUREK, ANDRZEJ KÜBLER and MICHAŁ ZIMECKI

    Abstract. The aim of this study was to monitor plasma interleukin 6 (IL-6) and tumor necrosis factor a (TNF-a ) levels in patients subjected to cardiac surgery under general anesthesia. In addition, proliferation of peripheral blood mononuclear cells (PBMC) to phytohemagglutinin (PHA) and the ability of these cells to secrete lipopolysaccharide-induced IL-6 and TNF-a during the observation period was investigated. IL-6 and TNF-a levels were measured using bioassays. We found that despite high variability in the postoperative response of the patients, characteristic kinetics in the appearance of the cytokines in plasma could be demonstrated. Most significant phenomenon was an increase of IL-6 level 1 day after operation associated with an inhibition of TNF-a concentration. Relatively high, preoperative concentrations of these cytokines were probably elicited by stress. Control, healthy donors, did not exhibit measurable levels of these cytokines. In terms of proliferative response of PBMC to PHA and of cytokine production in vitro, the patients could be classified into low and high responding. The proliferative response of PBMC from low responders was progressively increasing and the response of high responders did not exhibit meaningful changes throughout the observation period. PBMC from low producers of IL-6 showed also an increased ability to secrete this cytokine during the monitoring time while the cells from high producers yielded less IL-6 on the last day of the follow up. Regarding TNF-a production, PBMC from low responders reacted strongly by secretion of this cytokine on day 1 after surgery in contrast to high responders whose cells yielded less TNF-a after surgery. In majority of cases TNF-a production had a tendency to diminish on the last day of observation. We conclude that the changes in cytokine levels as well as an altered reactivity of PBMC in vitro reflect the immune response to the postoperative trauma and are desirable for a normal course of the immune response and wound healing.

    Keywords: cardiac surgery; peripheral blood mononuclear cells; proliferation; interleukin 6; tumor necrosis factor a.

Immunochemistry

  • Comparison of Serological Reactions of Rickettsiae-Infected Patients and Rabbit Anti-Proteus OX Antibodies with Proteus 0X2, 0X19 and OXK Lipopolysaccharides
    KEN-ICHI AMANO, MACIEJ CEDZYŃSKI, ANNA S. SWIERZKO, KAORI KYOHNO and WIESŁAW KACA

    Abstract. The reactivity of anti-Rickettsiae human antibodies with Proteus OX cells is used as a presumptive rickettsial diseases diagnostic test Weil-Felix reaction. In presented studies we compare the reactivity of human anti-Rickettsiae and rabbit anti-Proteus antibodies with series of Proteus OX2, OX 19 and OXK lipopolysaccharides (LPS). Polyclonal rabbit anti-OX2, anti-OX19 and anti-OXK sera reacted only with the homologous LPS – OX2, OX19 and OXK, respectively. The antibodies of Japanese spotted fever patients were less specific and reacted with OX2 as well as OX19 LPS. The antibodies of scrub typhus patients recognized Proteus OXK LPS, only. The serological reactions of O-antigen of P. mirabilis S1959 indicated that this, previously serologically not classified, strain may belong to the OXK group. Bacteria, used in the studies, came from the American, Japanese and European strain collections. The series of OX2, OX19 and OXK LPS, isolated from these Proteus strains, presented pattern of electrophoretic mobility and serological reactivities specific for each of OX – group types.

    Keywords: Proteus O-antigens serology; lipopolysaccharide, Weil-Felix test.il-Felix test.

  • Alterations of Tamm-Horsfall Protein Immunoreactivity after Partial Desialylation and Deglycosylation
    TERESA GRABSKA, TOMASZ BAGIŃSKI, ALEKSANDRA KUBICZ, MICHAŁ KOKOT, FRANCISZEK KOKOT, and JAN DUŁAWA

    Abstract. The role of the carbohydrate moiety of Tamm-Horsfall protein (THP) was investigated by studying the effect of partial enzymatic desialylation and deglycosylation on its immunoreactivity. In our experiments primarily sialic acid a 2,6 linked to galactose was split off, resulting in a 14% increase of immunoreactivity. It was increased up to 21 % by further removal of the sugar residues. We also found that complexes composed of THP and human serum albumin, immunoglobulin G or transferrin reacted with anti-THP antibodies. When these complexes were formed with desialylated or deglycosylated THP an essential increase of this reactivity (about 25%) occurred. Our studies indicate that modifications of the carbohydrate moiety of THP (which might occur in vivo by changes in the glycosylation process e.g. in pathologic conditions) lead to increased exposure of THP to the immune system.

    Keywords: deglycosylation; desialylation; enzyme-linked immunoassay; immunoreactivity; Tamm-Horsfall protein.

  • Selective Spectrophotometric Determination of Glucose and Fructose in the Presence of Aldoses Using Phenol-Acetone Reagent and Cerium(III) Chloride
    ANDRZEJ GAMIAN, JANUSZ BORATYŃSKI, M. ANNA ŻAL and RENÉ ROY

    Abstract. Aqueous mixtures of glucose and fructose produce red solutions when treated with 2%(w/v) phenol in 5% (v/v) aqueous acetone in the presence of concentrated sulfuric acid. The color is stable for days, and the red chromophore has an absorbance maximum at 568 nm. When the concentration of phenol is raised to 25%, fructose, but not glucose, produces red solutions, allowing for the selective detection of ketoses. Two complementary methods have been developed to remove the interference of ketoses in solutions containing glucose. The first one relies on the selective reduction of ketoses with sodium borohydride in the presence of cerium(III) chloride prior to the addition of the phenol-acetone reagents. The second method is based on the differential specific determination of glucose using 2% versus 25% levels of phenol. The relative sensitivities of different sugars are also presented as well as the applicability of the methods using bacterial polysaccharides for immunochemical analyses. The quantitative determination of glucose or ketoses in the polysaccharides does not require hydrolysis prior to the estimation.

    Keywords: glucose; fructose; sugar analysis; bioassay; glycoconjugate structure; spectrophotometry.trophotometry.

Experimental Therapy

  • Treatment of Drug-Induced Agranulocytosis with Colony Stimulating Factors (G-CSF or GM-CSF)
    JOANNA GÓRA-TYBOR, EUZEBIUSZ KRYKOWSKI and TADEUSZ ROBAK

    Abstract. The application of granulocyte-macrophage and granulocyte colony stimulating factors (GM-CSF and G-CSF has been progressively increased in the treatment of patients with agranulocytosis. The aim of our study was to compare the time of neutrophil recovery in patients with severe agranulocytosis treated with G-CSF or GM-CSF and the historical control group. We have studied 6 patients with agranulocytosis treated with stimulating factors and 7 patients in historical control group. Most of the patients have been exposed to thiamazole or non-steroid antiinflammatory drugs. Our results demonstrate that patients receiving colony stimulating factor have a significantly shorter period of recovery (the mean time 8.7 ± 1.98 days) than the historical control group (the mean time 11.0±2.24 days). We observed also a shorter time of antibioticotherapy and hospitalization in the group of patients treated with colony stimulating factor.

    Keywords: drug-induced agranulocytosis; granulocyte colony-stimulating factor; granulocyte-macrophage colony-stimulating factor.factor.

  • Interaction of 2-Chlorodeoxyadenosine in Combination either With Interferons or Recombinant Human Tumor Necrosis Factor a on Myeloid Progenitor Cells in vitro
    ANNA KORYCKA and TADEUSZ ROBAK

    Abstract. The objective of the present study was to investigate the interactions of 2-chlorodeoxyadenosine (2-CdA) with interferon a or g (IFN-a , IFN-g ), as well as between 2-CdA and recombinant human tumor necrosis factor a (rhTNF-a ),0n the clonal growth of granulocyte-macrophage progenitor cells (CFU-GM) from patients with chronic myeloid leukemia (CML) and on clonogenic leukemic blasts (CFU-L) from acute myeloid leukemia (AML) patients. Progenitor cell culture in semisolid medium in vitro was applied and the percentage of colony growth inhibition was evaluated. The use of 2-CdA either with IFN-a or IFN-g and 2-CdA with TNF-a was found to inhibit, in a dose dependent manner, the growth of colonies formed by hematopoietic precursor cells from CML and AML patients as well as from normal individuals, with the greatest effect being observed after the use of 2-CdA and IFN-a at their highest concentrations.

    Keywords: 2-chlorodeoxyadenosine; myeloid progenitor cells; recombinant human tumor necrosis factor a ; interferon a ; interferon g .

Various

  • Psoriasis Vulgaris with the Early and Late Onset- HLA Phenotype Correlations
    ANETA SZCZERKOWSKA-DOBOSZ, WALDEMAR PLACEK, ZOFIA SZCZERKOWSKA and JADWIGA ROSZKIEWICZ

    Abstract. The purpose of the study was to classify psoriasis vulgaris basing on HLA phenotype and the age of onset of the disease. One hundred fifteen psoriatic patients were included into the study and divided into two groups: group I – with early onset ( < 40 years) and group II – with late onset (  40 years). Each group was subclassified according to Cw6 antigen expression: IA-Cw6+ – early onset, IB-Cw6 – early onset, IIA-Cw6+ – late onset, IIB-Cw6 – late onset. HLA class I antigen typing was performed in each of 115 subjects using the microlymphocytotoxicity test. HLA class II antigens typing was also performed in 20 randomly selected patients by means of the two-step microlymphocytotoxicity test. The occurence Cw6, B 13, B 17 antigens was significantly increased in psoriatic groups in comparison with a control population. No difference between the groups was found with respect to clas II antigen frequency. Cw6, B 13 and B 17 were the most specific markers for psoriasis with the early onset, whereas B27, Cw2, B44 and Cw5 seemed to be associated with the late type of the disease.

    Keywords: HLA and disease; psoriasis vulgaris; relative risk.e risk.

  • Immunohistochemical Study in Breast Carcinoma. S100-Protein Positive Cells and Neuroendocrine Differentiation
    ALBINA ZÓŁTOWSKA, GRAŻYNA MOSZKOWSKA, BARBARA ZAMORSKA and ANDRZEJ ROSZKIEWICZ

Abstract. In this study S100-protein positive cells were found in all 12 cases of benign dysplastic changes and in all of the investigated 53 cases. of breast carcinomas. These cells belong to interdigitating cells (IDC) and were located in a regular pattern in the basal cell layer of the ducts and alveolar nodules in benign lesions. However, in breast carcinomas S100-protein positive IDC were in various pattern of distribution; only in the stroma, in basal cell layer, between cancer cells and S100-protein positivity of IDC and epithelial cancer cells. This phenomenon suggests the antigenic modulation of cancer cells transferring them the property of S100-protein positive cells or it may be the consequence of fusion. Immunoreactivity for chromogranin A (ChgA) and synaptophysin (Syn) was found in epithelial cells in some cases of benign dysplastic changes. However, in some carcinoma cases ChgA and Syn were revealed in carcinoma cells, in myoepithelial cells, in some stromal mesenchymal cells and endothelial cells. Immunoreactivity to smooth muscle actin was shown in the endothelial and smooth muscle cells of the vessel’s wall, in myoepithelial cells and was also sporadic in carcinoma cells.

Keywords: S100-protein; interdigitating cells; chromogranin A; synaptophysin.synaptophysin.

Vol. 44, No. 5-6, 1996

CONTENTS

Taxonomy

  • Present Status of Classification of Viruses of Vertebrates
    KONRAD MALICKI and FELIX N. TOKA

    Abstract. The VIth Report of the International Committee on Taxonomy of Viruses (ICTV) was published in 1995. We have briefly characterized its contents and discussed the most important changes that have been made in the classification of viruses of vertebrates. The present line up of families and genera of viruses of vertebrates, and subviral agents and unassigned viruses is also provided. We propose an allocation of families of viruses of vertebrates according to a conjectural evolutional connection between the type and class of genomic nucleic acid. Finally the directions and intentions of the ICTV have been reviewed from the Virology Division News in Archives of Virology (1994/1995). These will be considered in the VIIth Report of the ICTV scheduled for publication after the XIth International Congress of Virology in Sydney in 1999.

    Keywords: virus taxonomy; viruses of vertebrates.brates.

  • Hepatotropic Viruses Seen by Clinicians
    WŁADYSŁAWA ZIELIŃSKA and TOMASZ SMIATACZ

    Abstract. During last decades we have been witnessing substantial progress in hepatology, particularly in relation to viral hepatitis. Novel diagnostic tools allowed researchers to define new etiological agents, understand, their epidemiology and pathogenesis as well as develop effective preventive measures. However, several important questions still remain to be answered and no satisfactory therapeutical agents for chronic hepatitis are available. Basic epidemiological, preventive and clinical aspects of viral hepatitis are being discussed in this paper.

    Keywords: viral hepatitis; epidemiology; hepatitis A virus; hepatitis B virus: hepatitis C virus. virus.

Etiopathogenesis

  • Cytomegalovirus and Atherosclerosis
    JOSEPH L. MELNICK, ERVIN ADAM and MICHAEL E. DEBAKEY

    Abstract. The investigations reviewed in this paper provide evidence that a member of the herpesvirus family can cause atherosclerosis in chickens. In vitro experiments, as well as studies of arteries from infected birds, suggest that a virus-induced alteration of cellular metabolism, which results in the accumulation of cholesterol and cholesteryl esters, may be the primary mechanism in development of viral atherosclerosis. In addition, the fact that the same avian virus induces a malignant lymphoma suggests that it may also have the potential to stimulate the proliferation of other cells, notably arterial smooth muscle cells. The evidence for involvement of one or more members of the herpesvirus family in human atherosclerosis is much more circumstantial. Cytomegalovirus (CMV) is prevalent, increasing with age, so that a majority of the human population becomes infected by adulthood. As with other herpesviruses, the infection with CMV is usually subclinical or latent. Although the sites of latency for CMV have not been established, both smooth muscle and leukocytes are likely possibilites. The observations of viral antigens and nucleic acid sequences, but not infectious virus, in arterial smooth muscle cells suggests that latent CMV infection of the arterial wall may be common in patients with atherosclerosis.

    Keywords: atherosclerosis; cytomegalovirus.ovirus.

  • Characterization of Rotaviral RNA Isolated from Children with Gastroenteritis in Poland
    ZOFIA SZCZEPANIAK, WŁODZIMIERZ GUT and ZDZISŁAW JARZĄBEK

    Abstract. Rotaviruses were detected in 37% (331/895) of stool specimens collected from children with gastroenteritis attending three hospitals in Warsaw between January 1981 and March 1994. An analysis of rotavirus electrophoretypes circulating in Warsaw and Cracow, Poland, over 6 years showed that all 176 electrophoretypes encountered were characteristic of group A rotaviruses. Based on variations in the migration pattern of RNA genome segments when passed through polyacrylamide gels,10 different electrophoretypes were identified: 86% were long patterns and 14% were short patterns. Mixed infections were observed in two cases.

    Keywords: rotavirus; RNA electrophoretype; molecular epidemiology.iology.

  • Content of ß-Carotene in Blood Serum of Human Papillomavirus Infected Women with Cervical Dysplasias
    ANNA KWAŚNIEWSKA, ANNA TUKENDORF and MARIAN SEMCZUK

    Abstract. Studies were carried out in 528 women hospitalized in the Department of Obstetrics and Gynecology, Medical Academy in Lublin. Besides the control group, patients were classified according to the observed histopathological changes in the cervix (CIN) and found infections with human papillomavirus (HPV). In a11 cases ß-carotene content in blood serum was examined. HPV infection was probably a cause of decrease of ß-carotene content. It was found that with increased advancement of cervical dysplasia the level of ß-carotene in serum decreased.

    Keywords: human papillomavirus; cervical intraepithelial neoplasia; ß-carotene level. level.

  • Primary Hepatocellular Carcinoma in Patients with Chronic Hepatitis B Virus Infection
    HANNA KISZKIS

    Abstract. The paper presents 66 hepatocellular carcinoma (HCC) patients (57 males, 9 females) with chronic hepatitis B virus (HBV) infection in the clinical, diagnostic and therapeutic aspects. In 60 cases, hepatic cirrhosis was the primary hepatic disease, in 5 – chronic hepatitis, and in 1 – neoplasm developed in morphologically normal liver. Forty out of 66 patients were HBsAg seropositive. In 96% of the cases, anti-HBc antibodies were detected. In 76% of the cases, the infection was subclinical. In all patients, neoplasm was diagnosed in the late, symptomatic stage of the disease. Okuda’s classification was used to stage the disease. The following treatment methods were applied: surgical in 5 patients, embolization of hepatic artery with intra-arterial administration of cytostatics in 5, intravenous chemotherapy in 39, interferon a in lozenges in 15. In 44 patients, an objective response to treatment (according to WHO criteria) was achieved: in 5 (I and II Okuda’s stage) – a complete response (CR) for the period of 0.5-4 years, in 2 (III Okuda’s stage) – partial response (PR) with prolonged survival time up to 1 year, in 21 – no change (NC). The average survival time of non-treated and treated patients was, respectively: in the I Okuda’s stage 2.05 and 9.55 months, in stage II 1.36 and 7.36 months, in III stage 1.15 and 5.05 months. The differences in the average values are statistically significant. Only combined therapy, including both surgical and devascularization methods, applied to HCC patients results in achieving a long-lasting complete remission of the disease. Interferon a oral therapy improves the effects of the treatment.

    Keywords: hepatocellular carcinoma; HBV infection; clinical course of the disease; diagnostics; treatment; oral interferon a .

  • Acquirement of Susceptibility to Vesicular Stomatitis Virus Infection by Murine Resident Peritoneal Cells during Culturing in Vitro
    BEATA ORZECHOWSKA and ZOFIA BŁACH-OLSZEWSKA

    Abstract. We have studied the effect of culturing in vitro of BALB/c – mouse resident peritoneal cells (RPC) on their ability to replicate vesicular stomatitis virus (VSV). Results of 36 experiments performed on RPC, freshly isolated from individual female mice, showed that over 70% of them expressed constitutive antiviral immunity against VSV infection. The virus multiplied to low titers (? 103 TCID50/106 cells) only in 10 out of 36 cases. After several days in culture, the RPC were found to lose the resistance and VSV multiplied in the cells to high titers (up to 107 TCID50/106 cells). The time period, required for the cultures to reach a full ability to replicate the virus, was individually differentiated and ranged from 24 h to over 96 h.

    Keywords: peritoneal cells; in vitro culture; constitutive immunity; vesicular stomatitis virus.

Immunodeficiency

  • Syphilis and AIDS
    JADWIGA PODWIŃSKA

    Abstract. Evidence is shown that syphilis has become a major public health problem again. From 1988 to 1995 the permanently growing number of new cases of syphilis in the world was observed. The majority of the syphilitic cases in the patients are difficult for curing. The central nervous system is often involved in early syphilis. Previously the neurosyphilis was very rare. The reason for development of this stage of syphilis, may be an inadequate treatment as well as a weakening of the immunological responses. The latter one very often is caused by additional non-symptomatic infection including human immunodeficiency virus (HIV). Syphilitic ulcers act as a portal of entry for HIV. Analysis of cases with double infection with Treponema pallidum and HIV indicate that HIV infection may accelerate the course of syphilis and the presence of syphilis may also have influence on progression of the chronic HIV infection to AIDS. Taking into account that HIV infection alters the response to the treatment also, one can suggest that all of the patients with syphilis should be examined for the presence of HIV infection.

    Keywords: Treponema pallidum; human immunodeficiency virus (HIV). (HIV).

  • Does Hepatitis B Virus or Hepatitis C Virus Infection Influence CD4 Count in HIV-Positive Individuals?
    TOMASZ SMIATACZ and WŁADYSŁAWA ZIELIŃSKA

    Abstract. Objective of the study was to determine whether there is any influence of hepatitis B virus (HBV) or hepatitis C virus (HCV) infection on CD4 count decline rate in HIV-infected individuals. Retrospective analysis of consecutive CD4 counts was conducted in 72 HIV-infected untreated individuals (including 57 intravenous drug addict (i.v.d.a.)) in relation to their serological markers of HBV and HCV infection and the history of i.v. substance abuse. Anti-HBc seropositive individuals had slower CD4 count decline rates compared to anti-HBc seronegative ones (-0.20%/month vs. -2.90%/month, Kruskal-Wallis H = 4.77, p = 0.029). Anti-HCV serostatus had no influence on CD4 count decline rates (-0.47%/month for anti-HCV seropositive persons vs. -0.61%/month for anti-HCV seronegative ones, p = 0.91). History of i.v. substance abuse did not affect the CD4 count decline rates too (-0.83 for i.v.d.a. vs. +0.74 for non-i.v.d.a., p = 0.26). In our study HIV-infected individuals seropositive for anti-HBc tended to have substantially lower CD4 count decline rates compared to seronegative ones. Neither anti-HCV serostatus nor the history of i.v. substance abuse influenced the CD4 count decline rate. This observation arises question about the possible nature (molecular?, immune-based?) of potential mutual interactions between HIV and HBV infections.

    Keywords: hepatitis B virus; hepatitis C virus; human immunodeficiency virus; substance abuse; CD4; lymphocyte subsets.ocyte subsets.

  • Long-Term Assessment of Interferon Responses in the HIV + and AIDS Patients
    EGBERT PIASECKI, BRYGIDA KNYSZ, MAŁGORZATA INGLOT, KRZYSZTOF SIMON and ANDRZEJ GŁADYSZ

    Abstract. Impairment of interferon (IFN) system in human immunodeficiency virus (HIV) infection and acquired immunodeficiency syndrome (AIDS) became a basis for searching IFN responses to monitor the disease progression. For detailed investigations 16 HIV+/AIDS patients with at least 4 successively taken blood samples available for IFN determinations were selected. IFN responses were tested in two ways. Firstly, IFN level in plasma was measured. Secondly, capacity of IFN production by leukocytes was evaluated. The latter was determined in the whole blood assay, in which Newcastle disease virus (NDV) and phytohemagglutinin (PHA) were used as IFN-a and IFN-g inducers, respectively. The levels of IFN induced in whole blood leukocytes varied considerably in all individuals that had been tested. Nevertheless, two patterns of IFN responses were observed. In pattern I, patients had low levels of IFN in plasma and high levels of induced IFN-a and IFN-g . It was characteristic for 8 patients in good clinical condition. On the contrary, severe disease found in 2 patients was correlated with high levels of IFN in plasma and low levels of induced IFNs (pattern II). In 6 patients IFN responses were classified as intermediate pattern I/II suggesting transition from pattern I to pattern II. A variation of pattern I was found in the case of a patient defined as long-term survivor having relatively low levels of all IFN tested. The results suggested that interferon measurements reflected clinical condition of HIV+ patients showing not only past but also current immune changes.

    Keywords: interferons; interferon induction; HIV-positive patients; HIV progression marker.

  • Interferon and Tumor Necrosis Factor Production by peripheral Blood Leukocytes of Patients With Infectious Mononucleosis
    TERESA KAMIŃSKA, MARTYNA KANDEFER-SZERSZEŃ, AGNIESZKA SZUSTER-CIESIELSKA, GRAŻYNA RZESZOWSKA, HALINA MARKOWSKA and ROMANA MODRZEWSKA

    Abstract. Blood samples from 29 patients with infectious mononucleosis (IM) in phases of acute disease and convalescence were obtained. Interferon a (IFN-a ) and tumor necrosis factor a (TNF-a ) activity was detected in sera of patients both in: acute and convalescence phase, however when IFN titers were higher in the acute than convalescence phase, TNF titers were the highest in convalescence. In the whole blood assay Newcastle disease virus (NDV), phytohemagglutinin (PHA) and concanavalin A (ConA) and lipopolysaccharide (LPS) were used as cytokine inducers. A significant decrease in IFN titer induced in vitro with NDV, PHA and coon was observed in blood leukocytes of patients in the acute IM phase. In convalescence the ability of blood leukocyte of IM patients to produce IFN returned to normal, comparable with control. However, blood leukocytes of IM patients in the acute phase produced more TNF in response to LPs than in convalescence. The role of the observed overproduction of TNF in the course of IM similar to that in HIV infection should be elucidaeed.

    Keywords: infectious mononucleosis; interferons; tumor necrosis factor; human leukocytes.

  • Comparison of the Long-Term Effects of Treatment with Oral and Parenteral Interferon a in Chronic Viral Hepatitis Patients
    WŁADYSŁAWA ZIELIŃSKA, ANNA KORCZAK-ROGOŃ, JACEK PASZKIEWICZ, MACIEJ KOWALIK, JOSEPH M. CUMMINS and JERZY A. GEORGIADES

    Abstract. This report presents the interferon a (IFN-a ) treatment results for 75 patients with chronic hepatitis B virus (HBV) (51 cases) and hepatitis C virus (HCV) (24 cases) induced hepatitis in maximal 61 months follow-up. Among the group of 51 patients with chronic HBV hepatitis, 35 were treated orally with IFN-a in the form of lozenges in low daily doses (37.5 -150 U). The treatment was completed in 32 cases. The remaining 16 patients with chronic HBV hepatitis completed the treatment with parenteral IFN-a (3 x 106 U, 3 times a week). Positive results measured by the use of seroconversion in the HBe-antigen system were obtained for 68.7% (5-61 months follow-up) and 56.2% (7-44 months follow-up) of the patients treated with oral and parenteral IFN-a , respectively. Among the group of 24 patients with chronic HCV hepatitis, the first 6 patients were initially treated with IFN-a in the form of lozenges, in low daily doses. Biochemical remission was not achieved in these patients; genotype 1b was documened in 4 of them. Both, the first 6 patients (after a break) and the remaining 18 were treated with IFN-a parenterally, as in HBV patients. Temporary clinical and biochemical remission was achieved in 62.5% of the cases during the treatment, however the durable remission observed during 6 – 29 months of follow-up was achieved in 20.4 of the cases only.

    Keywords: interferon a ; interferon a in low oral doses; interferon a in parenteral use; hepatitis B virus (HBV) patients treatment; hepatitis C virus patients treatment.

Vaccines and Therapy

  • Viral Vaccines: Selected Topics
    MIROSŁAW KAŃTOCH

    Abstract. Significant role of viruses in pathology, their dominating position in etiology of infectious diseases point at the special position of active prophylactic procedures based on vaccination. The real role and value of viral vaccines of classic and modern generations, the limitation of immune potency in suppression of defence mechanisms, some problems of immunization against virus vertical transmission are presented in the paper. The reader may find tables which cumulate selected but significant patterns of viral vaccines and vaccinations, and selected papers devoted to topics discussed.

    Keywords: viral vaccines; immunodeficiency; vertical transmission.ission.

  • Controlling Orthopoxvirus Infections – 200 Years after Jenner’s Revolutionary Immunization
    MAREK G. NIEMIAŁTOWSKI, FELIX N. TOKA, ELŻBIETA MALICKA, MAŁGORZATA GIERYŃSKA, IRMA SPOHR DE FAUNDEZ and ADA SCHOLLENBERGER

    Abstract. An 11-year global WHO campaign for eradication of smallpox finished in October 1977 as the result of Edward Jenner’s primary success in 1796, who for the first time applied human vaccination against variola virus (VARV). The 200th anniversary of this happening is a good occasion to summarize the current status of the knowledge about the role of B and T lymphocytes in the control of orthopoxvirus infections. This short review concentrates on general characteristics of orthopoxviruses and the immune response to infection, mainly by vaccinia virus (VV) and ectromelia virus (EV).

    Keywords: orthopoxviruses; cellular and humoral immunity.oral immunity.

  • Studies on the Stability of HSV-1 Temperature Sensitive 28oC Clone
    BOGUMII.A LITWIIVSKA, AGNIESZKA BIESIADECKA and MIROSŁ,AW KAIVTOCH

    Abstract. The temperature mutants of herpes simplex virus type 1 has been received. The following properties on different levels of passages were investigated: stability of virulence, virus infection titer and immunogenicity. The virulence stability was expressed by percent of survivors and mean survival time in infected mice (CFW/Pzh and BALB/cPzh strains). Virus infection titer’s stability was measured after prolonged exposure of temperature sensitive mutant (ts 28oC) to critical temperatures (+4oC and from +20oC to +23oC). Stability of immunogenic potency of ts 28oC was evaluated in mice immunized with the virus on different level of passages and after that they were challenged with highly pathogenic temperature resistant clone (tr 39oC). The performed experiments proved the stability of investigated features.

    Keywords: herpes simplex virus; temperature mutants; stability.ts; stability.

  • Influenza : Problem in Poland as Elsewhere
    WIESŁAW MAGDZIK and LIDIA B. BRYDAK

    Abstract. The present paper deals with the epidemiological and clinical properties of influenza, taking into consideration differences in the course of the disease its consequences, and infections in high-risk groups. Special attention was paid to problems connected with influenza prophylaxis, and especially to the problem of vaccination. The authors point out that if the vaccine contains recommended strains for a given season and if the vaccination is done in the proper time, the immunity obtained in this way is effective.

    Keywords: epidemiology of influenza; influenza vaccination; immunity; vaccine.accine.

  • Characteristic of Humoral Response of Elderly to Inactivated Influenza Vaccine
    JOANNA SIENNICKA

    Abstract. Eighty persons immunized with inactivated influenza vaccine in 1990-1991 epidemic season were examined. Serological tests: hemagglutination inhibition (HI), lectin neuraminidase (LN) and Western blot were performed with serum specimens taken before and 3-4 weeks after vaccination. Distribution of anti-influenza antibodies in the subclass was also examined. The results revealed humoral response to vaccine strains of influenza virus and also to strains caused infections in the past.

    Keywords: influenza vaccination; elderly; humoral response.sponse.

  • Phosprenyl: A Novel Drug with Antiviral And Immunomodulatory Activity
    LEONID L. DANILOV, SERGEY D. MALTSEV, ANNA V. DEYEVA, ALEXANDER N. NAROVLYANSKY, ALEXANDER V. SANIN, SERGEY V. OZHERELKOV and ALEXANDER V. PRONIN

Abstract. A novel antiviral drug with immunomodulatory activity (Phosprenyl) is presented. The main active ingredient of the preparation is polyprenyl phosphates. This medicine is highly efficient against a number of viruses, including HIV in vitro, and tick-borne encephalitis and rabies viruses in the experimental models in vivo. In veterinary practice Phosprenyl is now regarded as an effective therapeutic means for treatment of canine distemper, hepatitis, enteritis, etc.

Keywords: Phosprenyl; polyprenyl phosphates; treatment of viral diseases.seases.