Mixed maternal-paternal lymphocyte cultures before and after immunotherapy for recurrent spontaneous abortions.

PROBLEM: The increased reactivity of maternal lymphocytes in reciprocal mixed-maternal-paternal lymphocyte cultures (MMPLC), observed in the presence of control serum after immunotherapy, suggests that immunization with paternal lymphocytes may induce a highly significant cell mediated immune response in specifically alloactivated maternal lymphocytes. METHOD: Reciprocal one-way MMPLC were set up with responding maternal or paternal lymphocytes and mitomycin C-treated stimulating lymphocytes. Cultures were set up for 6 days in the presence of 15% maternal or control serum. The degree of lymphocyte stimulation was measured by tritiated thymidine uptake. RESULTS: In maternal serum, after immunotherapy, a highly significant blocking effect on MMPLC was observed in both directions. The extent of the blocking effect in maternal serum and the stimulation in control serum was much higher, after immunotherapy, in two cases of abortions, as compared to cases with normal pregnancy outcome. CONCLUSIONS: Although the number of cases is very small, it may be that in abortions, in the presence of maternal serum, disturbances in the balance of cytokines or/and specific antibodies could have cytotoxic effects on MMPLC and down regulate, or "block" the specific response. For a possibly better utilization of the MMPLC test in the prediction of pregnancy outcome after immunotherapy, it may be important to examine specific antibodies in maternal serum, to investigate specifically induced cytokines in MMPLC and to evaluate T cell subsets in MMPLC in the presence of maternal and control serum.

Contribution of class I HLA-A2 antigen in immune reactions.

The involvement of the Class I HLA-A2 antigen is briefly reviewed in relation to allograft rejection, the feto-maternal relationship, viral cytotoxic reactions and tumor immunity. It is suggested that the HLA-A2 molecule may have, as compared to other HLA Class I alleles, a dominant role as a restricting element in cytotoxic T-cell recognition in the feto-maternal relationship to male fetuses, in specific viral infections and in tumors. As compared to other HLA Class I alleles, its reduced expression or loss in a variety of tumors suggests its possible important role in tumor immune surveillance. The disappearance of HLA-A2 from tumor cells may eventually contribute to the escape from T-cell recognition of malignant cells.

IN vitro cell-mediated immune reactions in herpes zoster patients treated with cimetidine.

In a double-blind placebo-control study the immunomodulating effect of cimetidine treatment for one week and placebo was investigated for cell-mediated immune reactions of 22 patients with herpes zoster (HZ). The mitogen induced leukocyte migration inhibition test (LMIT) and the in vitro proliferation of the patients' lymphocytes to exogenous IL-2 were used. Before any treatment, the mitogen induced leukocyte migration inhibition capacity (LMIC) of HZ patients was found to be significantly reduced (p < 0.02) as compared to healthy blood bank donors (controls). After one week, within the same treatment, the LMIC was significantly improved (p < 0.01). The patients' lymphoproliferative response to IL-2, before any treatment, was not significantly different from that of controls (p < 0.05). However, significantly higher values (p < 0.001) were found in patients tested 7 days after the disease onset as compared to those tested after 12 days. One-week cimetidine treatment significantly improved (p < 0.05) the lymphoproliferative response to IL-2 of initially low responders and had no effect on higher responder patients. In contrast to this, after one week of placebo treatment, a significant decrease in the patients' lymphoproliferative response to IL-2 could be observed as compared to patients' initial responses (p < 0.05) or to those of controls (p < 0.05). Although the number of cases is very small. The data suggest that after cimetidine treatment, as compared to placebo, healing from skin rash and pain was achieved in a significantly shorter time (p < 0.01).

Mode of inheritance of HLA haplotypes locus A,B in siblings of different sexes.

METHOD: Forty-eight parents and 172 children were typed for class I HLA antigens, locus A,B. RESULTS: Although the number of cases is small, we observed: (1) a significantly decreased number of sons born after a first delivery of a son, as compared to a first delivery of a daughter; (2) significantly increased sharing of maternal class I HLA antigens between the firstborn son and his brothers from higher birth orders, as compared to his sisters; and (3) HLA-A2 antigen, which is known to be involved in HLA restricted cytotoxic reactions in the recognition of minor histocompatibility antigens, was inherited in subsequent deliveries of sons as compared to daughters in a significantly higher frequency from the paternal than from maternal HLA haplotype. The results suggest that sharing of identical maternal HLA haplotypes between brothers may aid to decrease the degree of maternal sensitization to fetal antigens, and lack of HLA-2 antigen in maternal cells from sons as compared to daughters may avoid maternal HLA-A2 restricted cytotoxic reactions toward the male fetus.

Sialic acid level in maternal and neonatal lymphocytes and sera correlated to birth order and sex of the neonate.

Sialic acid (N-acetylneuraminic acid) was determined 1 h after normal term deliveries on peripheral blood lymphocytes from 42 mother-neonate pairs and in 29 maternal and neonatal sera. Results were evaluated according to maternal parity and sex of the neonate. The cases were divided into two groups: primiparae, and secundi- and multiparae. In primiparae the sialic acid level on lymphocytes from male neonates and from their mothers was by 23-30% decreased as compared to female neonatal and maternal cells. In the higher parity group, a significantly increased sialic acid level was found on lymphocytes from male as compared to female neonates, and maternal serum sialic acid concentration, unrelated to the newborns' sex, was by 17-20% increased as compared to primiparae. The results suggest that with increasing parity higher levels of sialic acid on male neonatal cells may possibly contribute to mask fetal male-specific histocompatibility antigens. Increased sialic acid levels in maternal sera from secundi- and multiparae suggest its possible contribution to an increased serum blocking effect.

Expression of transferrin receptor on human maternal and neonatal peripheral blood lymphocytes.

The levels of peripheral blood lymphocytes expressing the receptor for transferrin (TSR) on untreated and phytohemagglutinin (PHA)-stimulated cell samples from maternal-neonate pairs were evaluated 4-12 h postpartum. Significantly increased levels of TSR+ cells were observed on fresh, unstimulated neonatal and maternal cells, as compared to control cells from young adult males and females, and the values seemed to correlate with the sex of the neonate and with birth order. The level of TSR+ cells in culture was found to be increased on neonatal cells and decreased on maternal cells.

The effect of serum from patients with acute myocardial infarction on in vitro lymphocyte reactivity. II. Inhibition of IL-2 production.

Culture supernatants from concanavalin-A (con-A)-activated peripheral blood lymphocytes from healthy controls grown in the presence of sera from 20 patients 24 hours and 1 week after acute myocardial infarction (AMI) were tested for their mitogenic activity and for the presence of interleukin-2 (IL-2). Binding of exogenous IL-2 to activated lymphocytes from 10 patients was also determined. In supernatants prepared in the presence of patients' as compared to control sera, a significantly decreased mitogenic activity and IL-2 content were found. The mitogenic activity and IL-2 content in culture supernatants prepared with patients' sera collected 24 hours after the AMI (AMI I) and one week thereafter (AMI II) were significantly suppressed, and the degree of suppression in the 24-hour sera was significantly higher than in those collected after one week. No significant differences were observed in the binding capacity to exogenous IL-2 of activated patients' and control lymphocytes. The possibility is that immunosuppressive factors in the patients' sera, including cortisol, may suppress the patients' immune response acting through inhibition of IL-2 production.

The effect of serum from patients with acute myocardial infarction on in vitro lymphocyte reactivity. I. Inhibition of mitogen stimulation.

Sera from 20 patients obtained within 24 hours and one week after acute myocardial infarction (AMI) were tested for their immunomodulating effect on concanavalin-A (con-A) stimulated lymphocyte cultures from 11 healthy unrelated donors. Individual control sera from 21 healthy donors and 5 pools of control sera were used for comparison. Cortisol levels were tested in patients' and controls' sera. A significantly higher suppressive effect was seen in the presence of patients' sera taken at 24 hours than corresponding sera taken one week later. However, the suppressive effect after one week was increased as compared to control sera. A significant correlation between the degree of suppression and the cortisol level in corresponding sera was observed. An increased immunosuppression was observed with increased cortisol levels.

Role of children's sex in mixed mother-child lymphocyte culture reactivity.

One-way mixed mother-child lymphocyte cultures (MMCLC) 4 to 20 years after the last delivery were studied with maternal responding and children's or fathers' stimulating cells (MMFLC) in 14 multiple child families with 18 sons and 20 daughters. HLA antigen typing locus A, B, DR was performed for all family members. As reported previously for newborn cells, a significantly increased maternal response could be observed in MMCLC with male as compared to female children's stimulating cells. Although the number of cases studied was small, it seems that the increased stimulating effect of male children's cells could also be observed when MMCLC values from children of different sex, and identical A,B,DR haplotypes were compared. In contrast to this, A,B,DR haploidentical children of the same sex seem to have a similar stimulating effect on the maternal response in MMCLC. The results suggest that male children's Y-chromosome-correlated minor histocompatibility antigens may additionally stimulate the maternal immune response in MMCLC.

Changes in maternal and newborn lymphocyte reactivity in reciprocal mixed lymphocyte cultures during the postpartum period.

Reciprocal one-way mixed mother-newborn lymphocyte cultures (MMNLC) containing alternatively maternal or newborn responding (R) or stimulating (S) cells were investigated in both directions in primiparae at three different times: a few hours after delivery, and at 4 and at 16 weeks. Cultures were grown in the presence of maternal and pooled control serum prepared from the blood of five to eight unrelated healthy donors. Four weeks after delivery in maternal and in control serum a significant increase in MMNLC reactivity could be observed, which disappeared at 16 weeks when a pronounced decline in MMNLC values in both directions was found. The suppressive effect of maternal serum was more pronounced at delivery, still evident 4 weeks later, and insignificant after 16 weeks. The results of this study suggest that 4 weeks after delivery, maternal sensitization to fetal histocompatibility antigens can be detected in primiparae with MMNLC; and that 16 weeks later, this was no longer detectable with the same test.

Lymphocyte subpopulations in mother and newborn: correlation with sex of the newborn and number of pregnancies.

T cell subsets were defined with monoclonal antibodies of the OKT series, OKT3, OKT4 and OKT8, in 23 male and 22 female newborns and in their mothers 4-10 h after delivery. The data were compared and statistically evaluated between mother and newborn, between male and female newborns as well as between parity groups. The results indicate that the distribution of OKT4 and OKT8+ cells is different in mother and newborn and a significantly increased percentage of OKT4+ cells and a significantly decreased percentage of OKT3+ cells was observed in newborns as compared to their mothers after the first and second delivery. For maternal cells from male as compared to female newborns the percentage of OKT4+ was significantly decreased after the second delivery. OKT8+ cells in the mother were significantly decreased after the second as well as after three or more deliveries of male as compared to female newborns. With increasing parity the percentage of OKT3+, OKT4+ and OKT8+ cells decreased slowly for both sexes and the difference was significant between primi- and multiparae. The present findings suggest a possible role of the newborn sex and of parity in the distribution of specific T cell subsets in mother and newborn shortly after delivery.

Sialic acid content and sialyltransferase activity in human lymphocytes with advancing age.

Sialic acid and sialyltransferase activity were determined in lymphocytes obtained from the blood of 78 healthy male volunteers aged 20-80 years. When grouping was made in double decades, statistical evaluation using the Duncan procedure indicates that sialic acid did not show significant differences between groups, whereas the sialyltransferase activity was significantly higher in the group aged 41-60 years as compared to the group aged 20-40 years and the group aged 61-80 years, both at the 0.05 level.

Differences in sensitivity to melphalan between con A-activated and nonactivated human T-cell subsets.

In vitro treatment with melphalan (L-PAM, L-phenylalanine mustard), 2 micrograms/2 X 10(6) cells, significantly decreased the total number of E-rosette-positive (E+) T lymphocytes from peripheral blood (PBL) of healthy human donors as well as those of the OKT4 (precursor suppressor/helper/inducer T cells) and OKT17 populations (suppressor cells within the OKT4 subset). The OKT8 population (cytotoxic/mature suppressor cells) was not affected by a similar L-PAM treatment. The sensitivity of concanavalin A (Con A)-activated E+ T-cell populations to subsequent L-PAM treatment in vitro was different from that of Con A-untreated T cells: Thus, L-PAM treatment did not affect the expression of OKT3 and OKT4 antigens, increased the percentage of OKT17 cells, and inhibited the expression of OKT8 antigen. Depletion of OKT8 from Con A-activated E+ T cells (OKT4+-OKT8(-)-OKT17+) did not affect their suppressive activity on PHA stimulation in L-PAM-treated as well as untreated cells. Further depletion of OKT17+ cells from the OKT4+-OKT8(-)-OKT17+ subset (OKT4+-OKT8(-)-OKT17-) abolished the suppressive effect on PHA stimulation. Suppressive activity of the OKT4+-OKT8(-)-OKT17- subset was again evident after treatment of this population with L-PAM. The results obtained indicate that the sensitivity to L-PAM treatment of various T-cell phenotypes is changed by Con A activation and that after depletion of specific T suppressor cells L-PAM seems affect the immunoregulatory circuit within the Con A-activated OKT4 subset.

The immunomodulating effect of theophylline on lymphocytes from chronic lymphocytic leukemia patients.

The in vitro immunomodulating effects of theophylline on E-rosette formation, phytohemagglutinin (PHA) response, and Ig surface receptors of B lymphocytes were studied on fresh as well as on preincubated lymphocytes from patients with B cell chronic lymphocytic leukemia (CLL). In 11 out of 14 CLL patients, 24 hours preincubation at 37 degrees C significantly enhanced E-rosette formation. Subsequent treatment of preincubated cells with appropriate concentrations of theophylline further enhanced E-rosette formation in 11 cases. On fresh lymphocytes the enhancing effect of theophylline on E-rosette formation was not significant. The same was true for PHA stimulation; in 5 out of 7 cases the mitogen enhanced the stimulating effect of preincubation and had no significant effect on fresh lymphocytes from CLL patients. Preincubation significantly reduced the percentage of surface immunoglobulin positive B cells from CLL patients in all cases studied, and theophylline treatment had an additional effect on this phenomenon. No such effect of theophylline on fresh B cells from CLL patients could be observed. Preincubation had no significant effect on control lymphocytes. The effect of theophylline on control lymphocytes as compared to lymphocytes from CLL patients was completely different for T as well as for B lymphocytes. E-rosette formation from control lymphocytes (fresh and preincubated) was significantly inhibited in the presence of theophylline. No significantly enhanced responsiveness to PHA could be observed after treatment of fresh or preincubated lymphocytes with theophylline. Preincubation and theophylline treatment had no significant effect on the percentage of Ig positive B cells from control lymphocytes.(ABSTRACT TRUNCATED AT 250 WORDS)

In vitro selective effect of melphalan on human T-cell populations.

In vitro treatment with 2 micrograms/2 X 10(6) cells melphalan (L-PAM: L-phenylalanine mustard) significantly decreased the total number of T lymphocytes from peripheral blood (PBL) of healthy human donors and of the OKT4 population (precursor suppressor/helper/inducer) T cells as defined by monoclonal antibodies OKT3 and OKT4, respectively. No changes in the OKT+8 lymphocyte population (cytotoxic/mature suppressor cells) were observed following the same treatment. Preincubation of PBL with L-PAM at concentrations that do not affect the rate of DNA synthesis in PHA-stimulated lymphocytes inhibited the generation of T suppressor lymphocytes by ConA, as shown by their effect on PHA stimulation. Treatment of allogeneic PBL with L-PAM had no effect on mature suppressor T cells already induced by ConA, as shown by incubation of PBL with L-PAM after incubation with ConA.

Sialic acid in newborn and maternal lymphocytes: preterm deliveries.

Sialic acid was determined in newborn and maternal lymphocytes immediately after preterm deliveries of 34 and 35 weeks of gestation. The results were compared with those found in full-term deliveries. In contrast with full-term deliveries, in preterm deliveries a significant increase in sialic acid in maternal as compared to newborn lymphocytes was found. However, the mean cumulative value of sialic acid for maternal and newborn lymphocytes was similar in both groups. In addition to this, sialic acid concentrations seem to be sex-dependent and higher mean cumulative values for maternal and newborn lymphocytes could be found for male as compared to female newborns after full- as well as after preterm deliveries.