CONTENTS
Mini-Review
- Cytokines in the Treatment of Hematological Disorders: Recent Progress and Perspectives
TADEUSZ ROBAKAbstract. 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. GEORGIADESAbstract. 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 BŁASZCZYKAbstract. 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 LISOWSKIAbstract. 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 SZEWCZUKAbstract. 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 OCENASKOVAAbstract. 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 JADACHAbstract. 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. KAPPAbstract. 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 KAŃTOCHAbstract. 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 KLOCAbstract. 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.