CONTENTS
Cellular Immunology
- Participation of IL-2 and Direct Cell Cytotoxicity in the Modulation of GvH Reaction in Mice Infected with Listeria innocua
TERESA GOŚCICKA, MARIA WALENCKA, JACEK SZELIGA, MONIKA BLOCH and BOŻENA DZIADEKAbstract. Listeria innocua can intensify the development of local GvH reaction in a semiallogeneic system in mice. This phenomenon is observed in (BALB/c x AKR)F1 mice intraperitoneally injected with the live bac- teria on the 7th day before the transfer of BALB/c spleen cells. The local GvH reaction develops as strongly as in normal hybrid recipients given the parental cell graft in the presence of exogenous interleukin 2 (IL-2). The development of the reaction is associated with an intensive increase of the direct cytotoxicity of lymph node cells which is also markedly enhanced after the bacteria injection itself IL-2 seems to play a relevant role in the development of local GvH reaction in L. innocua-injected mice.
Keywords: GvH reaction; bacterial immunomodulation; Listeria.steria.
- Characterization of Cellular Components in the Peritoneal Fluid and in the Endometrial Tissue of Women with Endometriosis
KRZYSZTOF KAMIŃSKI, JAN KOTARSKI and MAREK GOGACZAbstract. The presence of leukocyte subpopulations was demonstrated in peritoneal fluid and in endometrial tissue of women with (n =16) and without (n =20) endometriosis. Peritoneal fluid samples were also assayed for effects on lymphocyte proliferalion in vitro. Peritoneal fluid leukocyte profiles were observed to be different between these groups. The most significant elevations in total number of leukocytes, macrophages and lym- phocytes and natural killer (NK) cells were observed in women with endometriosis. In normal eutopic en- dometrium T lymphocytes (CD3 ), macrophages (Ki-M 1 ) and NK cells (CD 16- CD56+) were present. In contrast aggregates of NK cells (CDl6+ CD56+) were only identified in ectopic endometrial tissue. Mito- gen-induced lymphocyte proliferation was significantly higher in the presence of peritoneal fluid from patients with severe stage of endometriosis as compared with other samples.0ur data indicate that disturbances of the cellular immune system may lead to progression of endometriosis in female peritoneal cavity.
Keywords: endometriosis; leukocyte subpopulations; peritoneal fluid; endometrial tissue.tissue.
- Presentation of Antigen by B Cell Subsets. V. Effect of Interleukin 7 (IL-7) and Interleukin 10 (IL-10) on Phenotype and Antigen Presenting Function of Immature B Cells
MICHAŁ ZIMECKI and JUDITH A. KAPPAbstract. In this communication we demonstrate that B cells from CBA/N mice with x chromosome-linked immunodeficiency and B cells from newborn mice, that have very limited ability to present antigen to an- tigen-specific T cell lines, acquire this function following preincubation with IL-7 or IL-10. These interleukins do not affect the antigen presenting function of B cells from normal, adult CBA mice. Incubation of B cells from xid and newborn mice with IL-7 or IL-10 but not with IL-2 induces expression of Lyb-5 marker on these cells. The study shows that the property of IL-7 and IL-10 to induce antigen presenting cell (APC) activity in immature B cells is associated with appearance of Lyb-5 antigen on these cells.
Keywords: antigen presentation; B cells; newborn mice; xid mice; interleukin 7; interleukin 10; Lyb-5 antigen.ntigen.
- CD4 + T Cells from Mice Primed for Humoral Immunity to Sheep Erythrocytes Specifically Inhibit Cell-Mediated Immunity and Vice versa
MICHAŁ ZIMECKI, JUDITH A. KAPP and ZBIGNIEW WIECZOREKAbstract. The aim of this study was to demonstrate whether development of the humoral or cellular immunity to sheep erythrocytes (SRBC) can be inhibited by a transfer of antigen-specific CD4+ T cells isolated from mice exhibiting cellular or humoral response. For generation of the humoral immune response and suppressor T cells for the cellular immunity, mice were immunized with a high dose of SRBC intraperitoneally (i.p.). On the other hand, for development of the cellular response and generation of T cells with inhibitory activity in relation to the humoral response, mice were immunized subcutaneously (s.c.) with SRBC emulsified with complete Freund’s adjuvant or intravenously (i.v.) with low dose of SRBC (105). We showed that depletion of CD4+ but not CD8+ T cells in the populations of transferred T cells abolished their inhibitory activity. The phenomenon of this suppression was dependent on a dose of T cells transferred and was antigen-specific, since T cells specific for chicken erythrocytes did not inhibit anti SRBC-response. The data indicate the importance of antigen specific CD4+ T cells in the mutual regulation of the two major type of the immune response.
Keywords: humoral response; cellular response; sheep red blood cells; CD4+; CD8+; T lymphocytes; sup- pression.ession.
Experimental Oncology
- Antitumor Activity of Bestatin and Thiorphan in Mice
JAN KOWALSKI, DARIUSZ BELOWSKI, ANDRZEJ MADEJ and ZBIGNIEW S. HERMANAbstract. We have investigated the effect of bestatin and thiorphan on growth of murine transplantable tumors, survival time, activity of natural killer (NK) cells and macrophages in mice. The injections of thior- phan at the doses of 0.5 or 5 µg per mouse retarded tumor growth and prolonged survival period in B16 melanoma bearing animals. Pretreatment with naloxone (an unspecific antagonist of opioid receptors) blocked the tumor growth inhibition induced by the treatment with bestatin and thiorphan what could suggest a contribution of endogenous enkephalin in this antitumor effect. The. percentage of mice bearing B 16 melano- ma tumor in the group treated with thiorphan at doses of 0.5, 5 or 50 µg per mouse was lower in comparison with control animals in a dose-dependent manner. The treatment with thiorphan at concentrations of 0.4-1.6 mg/ml inhibited growth of cultured in vitro B16 melanoma cells in comparison with control culture. Thiorphan added to the medium at concentration of 0.5 µg/ml or administered 4 times at the dose of 0.5 µg/mouse augmented NK lymphocyte activity.
Keywords: thiorphan; bestatin; tumor; natural killer cell activity; macrophages.phages.
- Peanut Agglutinin (PNA) Binding Glycoproteins on Human Urothelial Cell Lines of Different Grades of Transformation
DANUTA DUŚ, MACIEJ UGORSKI, WOJCIECH GORCZYCA and CZESŁAW RADZIKOWSKIAbstract. PNA-reactive sites, representing mainly unmasked Thomsen-Friedenreich (TF) antigen, are predic- tors of invasive transitional cell carcinoma. The present studies were undertaken in order: 1) to quantify the expression of PNA-reactive sites on well-characterized human urinary bladder cell lines belonging to two different transformation grades (TGr II and TGr III); 2) to identify PNA-binding glycoproteins that are restricted in their expression to tumorigenic and invasive urothelial cell lines. Flow cytomery studies revealed significant differences between TGr II and TGr III cell lines. The mean fluorescence intensities of TGr III, tumorigenic and invasive cells, were in the range of 28.4 to 57.3 arbitrary units. The TGr II cells exhibited several fold lower fluorescence intensity: 9.8 arbitrary units for HCV 29 cell line and 13.1 arbitrary units for Hu 609 cells. Neuraminidase treatment, increasing PNA-binding in TGr II as well as in TGr III cell lines, revealed the presence of cryptic PNA-binding sites. The number of cryptic PNA-binding sites seemed to be similar in TGr II and in TGr III cells and, therefore, only add to the total number of PNA-binding sites on native, untreated cells. Binding of 125 I-PNA to all cellular proteins resolved by SDS-PAGE and transferred to nitrocellulose showed multiple bands. The TGr III cell lines, after desialylation, were characterized by the presence of two major PNA-binding components represented by diffuse bands with apparent molecular mass about 68 – 79 k Da and 116 -156 k Da, respectively, and two weakly stained bands with apparent molecular mass of 51 kDa and 60 kDa. Both cell lines representing TGr II expressed high molecular mass PNA-binding component of 207 kDa. They were further characterized by the weaker staining intensity of 116-156 kDa glycoproteins as compared to TGr III cell lines.
Keywords: transitional cell carcinoma; transformation grade; Thomsen-Friedenreich antigen; peanut ag- glutinin binding; membrane glycoproteins.oteins.
- Growth Inhibition of Transplantable Tumors in Mice by mIL-2-Secreting Murine Plasmocytoma Cells Used Alone or in Combination with a Cytostatic Agent
ELŻBIETA PAJTASZ-PIASECKA, HALINA KUŚNIERCZYK, JAN SALWA, LESZEK KONARSKI and CZESŁAW RADZIKOWSKIAbstract. The non-tumorigenic cells of X63-Ag8.653 mouse plasmocytoma line tfansfected with murine inter- leukin 2 cDNA (X63-mIL-2) served us as the source of the cytokine to induce or to augment antitumor response in syngeneic BALB/c or semisyngeneic (CD2F1) mice challenged subcutaneously with either „wild” line tumor cells (X63/0) or with non-related methylcholantrene induced BFS1 fibrosarcoma of BALB/c mice. When applied peritumorally in several injections (2 to 6) to mice with non-advanced stages of the tumors, IL-2-secreting cells were able to cause tumor growth retardation in most of the treated mice and to induce tumor rejection in some of them. The combination chemoimmunotherapy was attempted in mice with advan- ced BFS1 fibrosarcoma using compound CBM-4A (the bromoanalog of ifosfamide) administered at various time (4 h or 3, 5 or 7 days) before the first of two local injections of transfected cells. The strategy proved to be more effcient in the tumor growth inhibition as compared with the cytostatic alone.
Keywords: murine tumors; murine interleukin 2 secreting cells; tumor vaccine; antitumor response; chemoim- munotherapy.herapy.
Immunopharmacology
- Influence of a Rat Kidney Perfusion Using the Anti-MHC Class II Monoclonal Antibody and Reduced Dose of 15-Deoxyspergualine on the Allogeneic Graft Survival
ANDRZEJ OKO, MACIEJ KRZYMAIŃSKI, ADAM DEJA, KARIN ULRICHS, ANA MARIA WAAGA and WOLFGANG MULLER-RUCHHOLTZAbstract. One of the main factors inducing rejection of the allogenic graft are the donor MHC-class II antigens. In this study, the allogenic rat renal graft survival after the blockade of MHC-class II positive cells was analyzed and compared with the effectiveness of the recipient treatment with 15-deoxyspergualine (IS-DOS). It was found that the DA (RT1 a) rat kidney perfusion with the anti-MHC class II monoclonal antibody (MoAb 29A1 – Kiel) allowed to prolong survival of the graft in the LEW (RT1 1) recipient (9.6±0.8 vs. 7.7±0.5 days). Our another study demonstrated that the 14 day treatment of the LEW recipient with 15-DOS at the dose of 0.5 mg/kg body weight can induce tolerance to the grafted kidney from the DA strain. The dose of 02 mg/kg body weight of 15-DOS prolonged the graft survival only to a small extend (16.5±0.5 days). In contrast, the combination of the graft pretreatment with MoAb 29A1 with the application of the reduced dose of 15-DOS to the LEW recipient allowed to further prolong the graft surwival (97.4±59.0 days, n = 5). In 3 cases, the long-time (close to 150 days) graft survival was obtained. The above presented results suggest that the blockade of the MHC-class II antigens can reduce the immunogenicity of the graft. Although this procedure was not sufficient to induce tolerance, it allowed to minimize the immunosuppressive treatment.
Keywords: experimental transplantation; organ perfusion; immunosuppressive treatment; monoclonal an- tibody; graft survival.rvival.
- Effect of Benzisoselenazolones and Organic Diselenides on Graft Versus Host Reaction and Immunoglobulins Levels in Chickens
BARBARA BLASZCZYK, ANNA D. INGNOT, PAAVO TOIVANEN, JACEK MŁOCHOWSKI and STANISŁAW SZYMANIECAbstract. Two week old chickens were treated once daily for 5 days with AE8 – I-pyridyl-1,2-benzisoselena- zol-3(2H)-one, AE22 – bis-2-(N-phenyle-carboxamido) 1 pyridyl diselenide and AE31 – bis(phenylo-diseleni- de with R3 = CONHC18H37). Their whole blood alone or blood mixed with thymus cells were used to generate graft versus host (GvH) reaction in 15 day old chicken embryos. The treatment of the chickens with the compounds stimulated the GvH reaction modifying activity of the donor cells as measured by increase of the spleen weight of the recipient chicken embryos. On the other hand, treatment with these compounds inhibited the IgG or IgM production in chickens immunized with human albumin.
Keywords: seleno-organic compounds; graft versus host reactions in chickens; immunoglobulins; enzy- me-linked-immunosorbent-assay.-assay.
- Immunotropic Activities of Benzisoselenazolones and Organic Diselenides in Mice
BARBARA BŁASZCZYK, ANNA D. INGLOT, STEFANIA H. KOWALCZYK-BRONISZ, STANISLAW SZYMANIEC and JACEK MŁOCHOWSKIAbstract. We have investigated the immunotropic effects of 23 seleno-organic compounds (8 benzisoselenazo- lones, 3 benzisoselenazolone oxides and 12 organic diselenides). All of the compounds increased the rosette formation of sheep red blood cells (SRBC) with spleen cells obtained from thymectomized CS3BL/6 mice and incubated in vitro in the presence of imuran. Furthermore,16 of the compounds were also assayed in vitro in the hydrocortisone test performed with CS7BL/6 mouse thymocytes. It was found that all of them significantly protected the cells against hydrocortisone induced cytotoxicity. Also in the Jerne’s assay, performed in 129Ao/ Boy mice pretreated in vivo with 3 selected compounds 5 days before immunization with SRBC, the stimula- tion of plaque forming cells (PFC) was observed. Only one compound (AE22, an analog of piroxicam) was found to be inhibitory in this assay. In contrast, in the graft versus host (GvH) assay performed in hybrid mice the donor lymphoid cells obtained from CS7BL/6 mice pretreated with 9 selected seleno-organic compounds, suppressed the GvH reaction in the recipient hybrid mice. Thus, in all of the immunotropic assays except the GvH reaction in adult mice, the seleno-organic compounds were found to have immunostimulating activities.
Keywords: seleno-organic compounds; hydrocortisone toxicity for thymocytes; rosette forming cells; plaque forming cells; graft versus host reaction in mice; immunostimulants.ulants.
- Oral Heparin in the Treatment of Rheumatoid Arthritis
JACEK IMIELA, JERZY NOSARZEWSKI and ANDRZEJ GÓRSKIAbstract. Heparin was administered orally in 10 patients with rheumatoid arthritis. The treatment resulted in clinical improvement in all cases (significant reduction in the number of tender and swollen joints and morning stiffness). This was associated with a decrease in erythrocyte sedimentation rate (8/10 patients) and cholesterol in all patients, although the differences were not significant. Recent data indicate that heparin can bind cytokines with potent immune modulatory action (e.g. TNF-a, interferon y etc.). Thus, neutralization of their action on target cells could be at partially responsible for the beneficial effects reported. Moreover, heparin inhibits T cell traffic to a site of antigen by blocking enzymes digesting the extracellular matrix and interfering with selection activity. The results of our preliminary study suggest that heparin, given per os, may have immunomodulatory properties with potential application in human disease.
Keywords: heparin; rheumatoid arthritis.hritis.
- 2-Chlorodeoxyadenosine (2-CdA) in the Treatment of Patients with Relapsed Chronic Lymphocytic Leukemia
EUZEBIUSZ KRYKOWSKI, KRZYSZTOF WARZOCHA and TADEUSZ ROBAKAbstract. This study attempts to characterize the response of patients with chronic lymphocytic leukemia (CLL) to the purine analog 2-chlorodeoxyadenosine (2-CdA). We have treated 10 patients with 2-CdA, at a dose 0.05 – 0.1 mg/kg/daily, for 7 days as a 2-hour infusion. Mean age was 54.6 years (range, 34 – 68 years). Mean time from diagnosis to treatment with 2-CdA was 37.0 months (range, 8 – 84 months). All the studied patients had received preliminary therapy consisting of other than 2-CdA chemotherapeutic regimens. Eight out of 10 patients had Rai stage III – IV disease. Four patients had Coombs positive hemolytic anemia before 2-CdA treatment. Seven patients responded to 2-CdA. Two complete remission (CR) and 5 partial remission (PR) were achieved. All patients but one with Coombs positive autoimmune positive hemolytic anemia achieved complete resolution of hemolysis. Severe neutropenia was frequent, and serious infections were noted in 20%, 43% and 50% of cases during the first, second and third course of 2-CdA, respectively. We conclude that 2-CdA is an effective agent in relapsed CLL patients, particularly in cases complicated by autoimmune hemolytic anemia.
Keywords: 2-chlorodeoxyadenosine; chronic lymphocytic leukemia; treatment.atment.
- Effect of Repeated Treatments with Cladribine (2-Chlorodeoxyadenosine) on Blood Counts in Multiple Sclerosis Patients
PAWEŁ GRIEB, ZBIGNIEW STELMASIAK, JANUSZ SOLSKI, JACEK NOWICKI, BEATA JAKUBOWSKA and MIROSŁAW RYBAAbstract. We report the results of blood morphology monitoring of 11 remitting-relapsing multiple sclerosis patients who received repeated treatments with cladribine (2-chlorodeoxyadenosine). The drug was given once, daily, subcutaneously (5 mg) or orally (10 mg) for 5 consecutive days, as 6 monthly courses followed by one or two additional courses at 3 or 6 month intervals. The treatments were well tolerated, although many patients suffered from incidental upper respiratory tract infections, most of which occured during the last 6 months of the observation period. One patient had recurrent infections, including an episode of urosepsis. All infections responded to standard therapy with antibiotics. Progressive lymphocyte reduction to 1000/µl on average, and clear but clinically insignificant drop in thrombocytes, was observed. Granulocyte counts were sometimes markedly elevated. A few patients developed macrocytosis, but none required transfusion. With our dosing and schedule, cladribine seems relatively safe in multiple sclerosis patients.
Keywords: 2-chlorodeoxyadenosine; blood counts; toxicity.xicity.
- 2-Chlorodeoxyadenosine: Lack of Synergism with Cyclosporine A and Tacrolimus (FK 506)
MARIA NOWACZYK, BARBARA STĘPIEŃ-SOPNIEWSKA, GRAŻYNA KORCZAK-KOWALSKA, TOMASZ MROWIEC and ANDRZEJ GÓRSKIAbstract. New clinically useful drug, 2-chlorodeoxyadenosine (2-CdA), was found to be a potent immunosupp- ressant both in vitro and in vivo. The present study examines the in vitro interactions of classical immunosupp- ressive agents – FK 506 and Cyclosporine A (CsA) with 2-CdA at the level of T and B cells proliferation, expression receptor for interleukin 2 (R-IL-2) and Ig synthesis. No synergism between 2-CdA and either FK 506 or CsA was demonstrable.
Keywords: 2-chlorodeoxyadenosine; cyclosporine A; tacrolimus; proliferation; receptor for IL-2; Ig synthesis.thesis.
- Continuous Versus Split-Course Irradiation for Lung Cancer. Immunological Implications
LJILJANA VUĆKOVIĆ-DEKIĆ, NEVENKA STANOJEVIĆ-BAKIĆ, MIODRAG DEKIĆ and OLIVERA FRIM
Abstract. The therapeutical irradiation for lung cancer causes profound disturbances of host’s general im- munocompetence, the cellular immunodepression being the dominant finding. It is thought that split-course technique holds certain advantage over the continuous irradiation, since the former includes an interruption of 4 week duration, thus allowing the lymphopoietic system to recover to a certain degree. In this report, we compared the radiotherapy-due alterations of several parameters of cellular immunity (the number and func- tion of total T cells, active T cells and the cells of monocyte/macrophage lineage), immediately after the completion of therapy in either continuously (n =13) or split-course-irradiated (n =12) lung cancer patients. All patients had received the total dose of 60 Gy. Both therapeutical techniques caused alterations of the parameters tested: the significant decrease of the total and active T cells and their proliferative responses, while the phagocytic activity and the number of mononuclear phagocytes were increased, the latter being affected to a lesser extent in split-course-treated patients. Our results suggest that both techniques have similar im- munodepressant effect on the cellular immunity of lung cancer patients.
Keywords: cellular immunity; iatrogenic immunosuppression; radiotherapy; lung cancer.cancer.