[Effect of estrogen on the blast transformation of lymphocytes and interleukin-2 production in lupus erythematosus]. / Oestrogen hatása a lymphocyta blastos transformatióra és az interleukin-2 termelésre systemás lupus erythematosusban.

The mitogenic response of peripheral lymphocytes was investigated in 12 patients with systemic lupus erythematosus and in healthy female volunteers who were on 11 and without 9 contraceptive pills. The effect of estrogen (ethinyl-estradiol 10(-5)-10(-6)-10(-7)M) was studied on Phytohaemagglutinin and Pokeweed mitogen induced blastogenic transformation and interleukin-2 production of peripheral lymphocytes in vitro. We observed a significantly depressed Phytohaemagglutinin induced lymphoblastic transformation both in patients and women taking oral contraceptive in presence of 10(-5)M estrogen as compared to normal controls. However there was no significant alteration neither in the response of lymphocyte nor in the production of interleukin-2 using of Pokeweed mitogen. The stimulataneous inhibition of the interleukin-2 production proved to be moderate. Marked significant correlation (r greater than = 0.8) vas detected between lymphoblastic transformation and interleukin-2 production in healthy females. Correlation coefficient measured in females taking oral contraceptive (r less than = 0.64) and patients with systemic lupus erythematosus (r less than = 0.34) suggest that in these groups the inhibition of lymphoblastic transformation is due to the inhibition effect of estrogen on the interleukin-2 production.

PIP: 12 patients with systemic lupus erythematosus (SLE) (8 in the active stage) with an average age of 26 years (17-54) and 20 healthy control subjects (9 were aged 18-49 years and 11 were oral contraceptives [OC] users aged 17-44) were studied to assess the inhibiting effect of estrogen in vitro on phytohemagglutinin (PHA) and Pokeweed (PWM) mitogen induced blast transformation of lymphocytes (lymphoblastic transformation=LBT) gained from periphral blood and simultaneous interleukin-2 (IL-2) production. 8 women were taking Anteovin, 2 Ovidon, and 1 Rigevidon. The average duration of OC use was 5.2 years. 1 SLE patient did not need immunosuppressive treatment, 3 patients received corticosteroid maintenance therapy, and 4 patients were also taking 50 mg of Imuran. In 4 active SLE patients the tests were done before raising the dose of immunosuppressive drugs, and in the case of 2 other patients the administration of 75 mg and 25 mg/die Prednisolone was necessary in addition to 50 mg and 100 mg/die Imuran. LBT decreased significantly in patients and OC users. The LBT values induced by PWM were similar but not significant. The IL-2 production induced by PHA decreased in all 3 groups but not significantly. I1-2 production was 6 E/ml in patients, 5 E/ml in OC users, and 11.5 E/ml in nonusers, but the differences did not prove significant because of wide individual fluctuations. The amount of IL-2 produced by lymphocytes at PWM stimulation was almost the same in all 3 groups with or without estrogen. There was a positive, significant relationship between the extent of LBT and the amount of IL-2 produced in the healthy group of nonusers, it was less solid in the OC users, and in the SLE group trhe low correlation coefficient of .34 suggested the reduction of IL-2 through the inhibition of LBT.

Effect of conditioned media of acute myeloid leukemia blast cells on complement synthesis by cultured human cells of monocyte and hepatocyte origin.

The effect of conditioned media of 3-day cultures of blast cells from peripheral blood of 5 patients with acute myeloid leukemia (CM-AML) was studied on the synthesis of C2, factor B (Bf) and C1 esterase inhibitor (C1-INH) by human monocyte-macrophage cultures and HepG2 hepatoma cell line. The level of C2 in the culture supernatants was measured by immune hemolysis, those of Bf and C1-INH by ELISA. CM-AML was added to the monocyte cultures on day 3 and replaced by culture fluid on day 6. Compared to the control cultures, CM-AML significantly increased C2 and Bf levels and slightly decreased C1-INH levels in the culture fluids on day 6. On day 9, Bf synthesis enhancement still could be observed but C2 and C1-INH levels did not significantly differ from those of the control. CM-AML significantly increased the synthesis of factor B by the HepG2 cells too. A strong correlation was found between the results of the Bf protein and RNA determinations, which means that the supernatants of AML blasts affect the gene expression of factor B at a pretranslational level. The selective complement synthesis modifying effect of CM-AML was not due to interferons (IFN) because neither IFN-alpha nor IFN-gamma could be detected in these conditioned media. The present findings indicate that the hypercomplementemia observed in AML patients can be due to unknown factor(s) produced by leukemic blast cells.

Non-cytotoxic blocking antibodies and suppressor cells induced by donor-specific transfusions in healthy volunteers and potential kidney transplant recipients.

Donor-specific transfusion (DST)-induced immunosuppression plays a significant role in clinical and experimental transplantation. To clarify the mechanism of suppression on alloreactivity the suppressor cell induction and the non-cytotoxic blocking antibody production and importance was studied in 15 healthy volunteers and 3 kidney transplant recipients (KTR) after DST on mixed lymphocyte culture (MLC). Significant decrease of anti-donor MLC response was found in all of KTR and in 12 cases of the 15 transfused volunteers. Of 18 cases, 15 had blocking factors in their post-DST serum which strongly (52-91%) inhibited the MLC response. The IgG fractions isolated from the "blocking sera" were responsible for this inhibition. Eighty-one percent of non-cytotoxic blocking antibodies affected the responder cells in MLC and reacted with third party responder cells as well. Both buffy coat and platelet transfusions evoked production of the non-specific blocking antibodies. Our data strongly suggest that DST induces not only the differentiation of suppressor cells and production of anti-idiotypic antibodies, but also the appearance of non-specific, non-cytotoxic antibodies, which may participate in the cell-mediated immune suppression of the alloimmune reactivity.

Enhancing effect of alpha-interferon on the C2 synthesis by cultured human monocytes.

C2 synthesis by human peripheral blood monocytes cultured in the presence of human alpha-interferon (IFN-alpha) was studied. IFN-alpha was added in different amounts (1-1000 IU/ml) to the cultures on day 3 and was removed on day 7. As control, mock interferon (m-IFN) was also tested. C2 content of the culture supernatants was measured by immunohaemolytic method. IFN-alpha was found to enhance C2 synthesis in a dose-dependent way. The enhancing effect could be observed even after the removal of IFN-alpha. C2 production by the cultured monocytes was increased by m-IFN as well; the extent of enhancement however, was found to be significantly lower than that induced by the corresponding amount of IFN-alpha. In contrast to the enhancing effect on C2 production, IFN-alpha did not influence total protein synthesis in the cultures, suggesting a selective stimulatory action on the C2 gene.

Independence of depressed lectin-dependent cell-mediated cytotoxicity from interleukin 2 production in patients with systemic lupus erythematosus.

The relationship of lectin-dependent cell-mediated cytotoxicity (LDCC) to interleukin-2 (IL-2) production was studied in healthy subjects and in patients with systemic lupus erythematosus (SLE). Profoundly depressed levels of LDCC were elicited by peripheral blood mononuclear cells (PBMC) from nine patients with active SLE in comparison to LDCC from seven controls, and eleven inactive SLE donors, using 3H-TdR-prelabelled adherent HEP-2 cells as targets in a 24 h assay with 25 micrograms/ml Con A. In parallel experiments, no individual correlation was found between LDCC activity and IL-2 production for healthy or SLE subjects. Further, no major differences were detected in IL-2 release when the three groups of donors were compared, a tendency observed at the Con A doses (5 and 25 micrograms/ml) and incubation times (24, 48, and 72 h) used to induce IL-2 production. In additional studies, impaired Con A-induced blastogenesis was noted for PBMC from active SLE patients in comparison to the PBMC from the controls or patients with inactive SLE. While strong individual correlation was obtained between blastogenesis and IL-2 secretion in controls and patients with inactive SLE, no such relationship was found in patients with active SLE. While addition of exogenous IL-2 to the cytotoxicity assay considerably enhanced LDCC by healthy donors it failed to improve LDCC by patients with active SLE. These data suggest that depressed LDCC and Con A-induced blastogenesis of patients with active SLE may not be related to impaired IL-2 production but rather to an inherent dysfunction of the effector lymphocytes, including their unresponsiveness to IL-2.

Lectin-dependent cell-mediated cytotoxicity and blastogenesis by large granular-enriched and depleted lymphocytes.

The role of larger granular-enriched and depleted lymphocytes was studied in lectin-dependent cell-mediated cytotoxicity (LDCC) against adherent HEp-2 human epipharynx carcinoma target cells. LDCC was evaluated by detachment from the monolayer of 3H-thymidine-prelabelled HEp-2 cells in a 24 h assay at effector-target cell ratios of 25:1 and 50:1 in the presence of 25 micrograms/ml concanavalin A (Con A). Under the aforementioned conditions but in the absence of Con A natural cell-mediated cytotoxicity (NCMC) was not found. However, cytotoxicity was significantly augmented in the presence of Con A (= LDCC) using human peripheral blood mononuclear cells (PBMC) as effectors. Large granular lymphocytes (LGL), which show high natural killer (NK) activity to K 562 target cells, failed to be cytotoxic against HEp-2 targets similar to large granular depleted lymphocytes (LGL-DL). On the other hand, LGL caused only a slight LDCC; whilst LGL-DL induced strong LDCC activity towards HEp-2 targets. In comparison to LDCC using LGL-DL as effector cells, LGL and LGL-DL mixed at a ratio of 1:2, and added to target cells, had no major effect on LDCC, while a lower level of LDCC was observed at LGL/LGL-DL ratios of 1:1, and 2:1, suggesting the dilution of LGL-DL, potential effectors of LDCC to HEp-2 cells, rather than a specific regulatory role of LGL in LDCC. In parallel studies, the proliferation of LGL-DL in response to Con A was less than that observed with PBMC or LGL. The response could be restored by replacing half of LGL-DL per culture with an equal number of LGL, or by the addition of 10% monocytes. Significant functional differences between LGL and LGL-DL in LDCC as well as in Con A-induced blastogenesis are suggested.

Mixed lymphocyte cultures (MLC) and acute pancreatitis after renal transplantation.

MLC was performed preoperatively in 35 out of 57 candidates for renal transplantation. Three patients developed acute pancreatitis in the early postoperative stage, 2 of them died. In these three cases MLC revealed complete non-reactivity of the recipient lymphocytes, compared with the donor as well as with the positive control lymphocytes. None of the other patients showed this phenomenon and none of them developed acute pancreatitis. It is suggested that the recipient's immune responsiveness may be involved in the aetiology of the production of acute pancreatitis after renal transplantation.

The natural killer activity (cytotoxicity) of lymphocytes.

The authors' own investigations into the spontaneous lymphocyte-mediated cytotoxicity (SLMC) have been reviewed and discussed in th light of literary data. It is suggested that SLMC is a resultant of complex effects contributed by antibodies present in the human serum and those attached to the surface of lymphocytes as well as by spontaneous (sui generis) functions of effector cells. The lymphocytes eliciting SLMC belong to the "O" subpopulation, bear Fc receptors and are presumably, identical with "K" lymphocytes. In animal experiments, the authors have shown that the SLMC reaction directed againt a virus-induced tumour is under polygenetic control primarily governed by gene(s0 linked to the histocompatibility region. In man, similarly as in the mouse, SLMC was found to be a genetically-controlled lymphocyte function in which gene(s) linked to the HLA-A2 B12 or the HLA-A3 B7 haplotype may play a determining role. The authors' clinical observations indicate that LSMC represents an important part of the defense mechanism against malignancies and autoimmune diseases. Estimation and follow-up of SLMC may be useful in monitoring the clinical course.

Changes induced by vascular antigens in the aorta of guinea-pigs: immunological and morphological studies.

The effects have been studied of human vessel wall on the cell-mediated and humoral immune response, as well as the morphological changes produced in the aorta in guinea-pigs. In animals immunized with calcium chloride-tris-citrate extracts of the aortic and vessel wall a definite cell-mediated and humoral immune response was observed. A cross-reaction was found between the 2 vessel wall extracts in the skin test and in the migration inhibition test. The induction of desoxyribonucleic acid synthesis was found to be specific. Antibody production as well as the concentration of IgG in the serum was increased. Simultaneously with increased production of the antibodies the cellular immune response did not diminish util the 9th week. A definite alteration was found in the aortic intima by histological, histochemical, immunofluorescence and electron microscopic methods. In addition some slight changes could be observed in the media and in the aventitia of the aorta.

Immunological studies with aortic and venous tissue antigens. III. Examination of cell-mediated and humoral immune response induced by human vascular antigens.

The effects of human vascular antigens on the cell-mediated and the humoral immune response were studied in guinea pigs. In animals immunized with buffer extracts of aortic and venous wall a well measurable cell-mediated and humoral immune response were developed. Between the two kinds of vascular extracts in the skin tests, migration inhibition tests and antibody-production tests a cross-reactivity was observed, whereas the induction of desoxyribonucleic acid-synthesis proved to be specific. No cellular or humoral immune response to vascular extracts was detectable in the control animals. Analysing the time-kinetics of the cell-mediated and humoral immune response it was found, that with the increased production of antibodies the cellular immune reactions did not cease until the ninth week.