CONTENTS
Mini-Review
- TNF Receptors – How They Function and Interact.
JACEK BIGDA and HELMUT HOLTMANNAbstract. 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. GRZESIKAbstract. 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 KÜBLERAbstract. 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. MACKIEWICZAbstract. 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 PODCIECHOWSKIAbstract. 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 HAŁASAAbstract. 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. ZHYTKEVITCHAbstract. 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 STRZĄDAŁAAbstract. 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 MAZURIERAbstract. 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 PŁYTYCZAbstract. 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 SZEWCZUKAbstract. 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. SIEMIONAbstract. 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 RADZIKOWSKIAbstract. 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.