Mechanism of action of maternal serum on the interleukin2 receptor expression.

Neoplastic Jurkat cells were submitted to a PHA stimulation test after a preincubation in maternal or nulliparous serum (10% dilution). The Il2R expression was significantly downregulated among maternal serum treated cells. Retroplacental serum was significantly more inhibitive than peripheral maternal serum (P less than 0.01). The maternal IgG fractions and mostly the retroplacental IgG fraction proved to contain a factor mainly responsible for the Il2R expression inhibitive property. The molecular mechanism of this phenomenon was further studied. It was shown that H7 (acting as a protein kinase inhibitor) could not influence the Il2R modulation. E.G.T.A., a calcium chelator, was not able to interfere with the inhibitive influence of maternal serum. It was suggested that the maternal serum mediated inhibition of the IL2R expression is not influenced by the hydrolysis of membrane bound phosphatidyl inositol. In contrast, pertussis toxin markedly enhanced, in a dose dependent way, the suppressive influence of maternal serum as compared to nulliparous serum. At low concentrations, pertussis toxin lost its stimulating property and retained its ability to ADP ribosylate the alpha subunit of G proteins inducing a release of adenylcyclase mediating cAMP synthesis. This mechanism has been further studied by the addition of dbc AMP or dbc GMP to Jurkat cells preparations stimulated by PHA. dbc AMP, in a dose-related way, induced a downregulation of the IL2R expression of stimulated neoplastic cells preincubated in nulliparous or maternal serum. dbc GMP did not influence the IL2R expression in the same experimental conditions. The maternal serum mediated cells showed the most pronounced IL2R inhibition. Finally, it was shown that the cAMP synthesis by PHA stimulated Jurkat cells was upregulated in a dose dependent way, after a previous cellular incubation in progressive concentrations of maternal serum. In contrast, among nulliparous serum pretreated cells, cAMP synthesis remained significantly lower, after a lectin stimulation, as compared to the cAMP production derived from retroplacental serum treated and stimulated cells. Taken together, these experiments suggest that the maternal serum dependent suppression of the IL2R expression is related to a protein G stimulation followed by an enhanced cAMP synthesis.

Modulation of B cell stimulation by maternal serum.

Anti-IgM stimulation of B cells is decreased in the presence of maternal serum as compared to control media. This inhibiting influence of maternal serum is observed during the priming of the B cells. The progression of B cells into cellular proliferation was not influenced by maternal serum. At the level of the immunoglobulin secretion, the influence of maternal serum was also shown. A significant down regulation of the IgM, no change of the IgG production, and an enhanced secretion of IgA and IgE was demonstrated in the presence of maternal serum as compared to control media. It has been suggested that the maternal IgG fraction contains a molecule partly responsible for these changes. Furthermore, the CD23 antigen is increased when B cells are stimulated in the presence of a pool of maternal IgG. All the findings concerning maternal IgG were more pronounced when retroplacental IgG was used instead of peripheral maternal IgG. This observation suggests that the factor responsible for the B cell changes is released at the fetomaternal interface.

Modulation of the HLA class II antigen at a molecular level by maternal serum among cord blood cells and unrelated lymphocytes.

A retroplacental serum factor responsible for the downregulation of the MHC class II antigen expression has been described [1]. We found that maternal serum had the same property. The HLA class II modulating activity is however diminished in the presence of maternal serum as compared to retroplacental serum suggesting that the IA like inhibiting factor is released at the fetomaternal interface. After a 3-day incubation period of unrelated lymphocytes and mononuclear cord blood cells in a maternal serum pool, it was shown that the HLA Dr, the HLADp, and more significantly, the HLADq molecules were modulated. This phenomenon was more pronounced among cord blood cells. When third party lymphocytes and mononuclear cord blood cells were stimulated by Candidine or unrelated mononuclear cells in the presence of retroplacental serum, only the cellular subpopulation belonging to the CD4+ subset showed an HLADq downregulation. The molecular constituents of the MHC class II antigen expression characterizing cells belonging to other subsets remained unchanged. When the same stimulation assay was performed in the presence of a control medium (nulliparous serum), we found no changes in the MHC class II molecular constituents. When unrelated mononuclear cells and mononuclear cord blood cells were PHA stimulated in the presence of maternal and nulliparous serum, the HLADq expression of the CD8+ subset showed a significant downregulation in the maternal serum mediated stimulation assay as compared to the control stimulation test. The molecular expression of the HLA class II antigen related to the other subpopulation (CD4+, CD3+) stimulated by a mitogenic lectin remained unchanged. It is suggested that these molecular MHC class II modulations are due to a factor included in the maternal IgG reaction. Retroplacental IgG contains the highest concentration of this factor.

The interleukin-2 receptor on cord blood and nulliparous mononuclear cells is downregulated by maternal and cord blood serum.

The concentration of interleukin-2 receptor (IL-2R) on maternal, cord blood and unrelated nulliparous mononuclear cells has been studied after a previous phytohaemagglutinin (PHA) stimulation. Furthermore, the inhibitive action of maternal and cord blood serum on the IL-2R expression of stimulated nulliparous and cord blood lymphocytes has been shown. A downregulation of the [3H]-thymidine uptake by these PHA treated cells previously incubated in maternal and cord blood serum has been observed. Retroplacental serum was the most inhibitive experimental medium. The IL-2R modulation property of maternal serum has been studied during the pregnancy. Appearing quite early (in the sixth week), the maternal serum dependent inhibitive factor vanished, 2 to 3 weeks after delivery. After a study of the different serum components, it is suggested that the IL-2R downregulating molecule is included in the maternal immunoglobulin (IgG) fraction. Further experiments suggest that the addition of recombinant IL-2 during the action of low doses of maternal IgG allows a partial re-expression of the IL-2R. However, at physiological concentrations of IgG, the IL-2R downregulation becomes irreversible.

Modulation of the interleukin 2 receptor by maternal and cord blood serum.

In this report, the potential inhibitive action, of maternal serum (retroplacental and peripheral), and cord blood serum on the expression of the Il2r has been studied. Calf and male serum were used as a control. In order to have an optimal Il2r expression, a previous PHA cellular stimulation was performed. The maternal serum's Il2r inhibitive property was measured during and after the pregnant period. The presence of Il2r on maternal lymphocytes, cord blood cells, and unrelated donor mononuclear cells has been investigated (after a PHA stimulation assay). The downregulated Il2r expression of neoplastic cell line (Hut78 cell line) under the influence of maternal serum has been observed. The examination of the inhibitive action due to maternal serum has suggested that a factor included in the IgG fraction is mainly responsible for the downregulating property concerning the Il2r expression. A possible mechanism for the action of this factor has been studied. Further experiments suggest that the addition of recombinant Il2 during the action of low doses of maternal IgG allows a partial reexpression of the Il2 receptor. However, at physiological concentrations of IgG, the Interleukin 2 receptor downregulation becomes irreversible.

Modulation of the HLA class II antigen at a molecular level by maternal serum.

The recent immunological literature has described the existence of a retroplacental serum factor being responsible for the downregulation of the MHC Class II antigen expression. In this report, the same property has been found in peripheral maternal serum. The HLA Class II modulating activity is however diminished in the presence of peripheral maternal serum as compared to retroplacental serum suggesting that the IA like inhibiting factor is released at the fetomaternal interface. After a three-day incubation period of unrelated lymphocytes in a maternal serum pool, it was shown that the HLA Dr., the HLA Dp., and more significantly, the HLA Dq. molecules were modulated. When third party lymphocytes were stimulated by Candidine or unrelated mononuclear cells in the presence of retroplacental serum, only the cellular subpopulation belonging to the CD4+ subset showed an HLA Dq. downregulation. The molecular constituents of the MHC Class II antigen expression characterizing cells belonging to other subsets remained unchanged. When the same stimulation assay was performed in the presence of a control medium (nulliparous serum), no changes concerning the MHC Class II molecular constituents were observed. When unrelated mononuclear cells were PHA stimulated in the presence of maternal and nulliparous serums, the HLA Dq. expression of the CD8+ subset showed a significant downregulation in the maternal serum mediated stimulation assay as compared to the control stimulation test. The molecular expression of the HLA Class II antigen related to the other subpopulations (CD4+, CD3+) stimulated by a mitogenic lectin remained unchanged. It is suggested that these molecular MHC Class II modulations are due to a factor included in the maternal IgG reaction. Retroplacental IgG contains the highest concentration of this factor.

Functional modifications of the CD4+ and the CD8+ subset due to maternal serum.

Numerous experiments have been performed to try to explain the successful gestation of the semiallogeneic mammalian fetus in the immuno-competent mother. A popular hypothesis is that localized intra uterine suppression mediates the immune response and contributes directly to the survival of the fetus. Suppressive factors synthesized at the fetomaternal interface may be transported by the bloodstream and be found in the retroplacental and peripheral blood circulation. In this report we aim to study these modulating factors by exposing a proteinaceous antigen (Candidine or human mono nuclear cells) to unrelated human lymphocytes in the presence of a pool of maternal serum, retroplacental (MAT SR) or peripheral (MAT SP), or to a pool of male or calf serum (MALS or CS). A significant downregulation of the candidine mediated lymphocytic stimulation was observed in the case of the maternal serum. In order to further characterize this inhibition, a quantitative evaluation of the expression of the CD4 and CD8 positive subpopulations was performed. A selective inhibition of the CD4 positive subset was observed. In the PHA stimulation assay in the presence of maternal serum there was an inhibition of the thymidine uptake of unrelated lymphocytes. When studying the different subsets which were stimulated in the presence of maternal serum and control media it was shown that the CD4 positive subpopulation remained unchanged while there was a slight inhibition of the CD8 positive subset in the first case (maternal serum treated). The same CD4 positive inhibitive property of maternal serum was observed when a neoplastic cell line (HUT cells) was used as target for candidine in a stimulation test. This CD4 inhibited expression remained constant as long as the maternal serum was renewed. Maternal lymphocytes however remained resistant to the inhibitive action of maternal serum and did not show any change in their CD4 and CD8 positive subpopulation. By investigating the different blood components, it was shown that the suppressive factor was included in the IgG fraction and synthesized at the placental level. Appearing quite early during the gestation (at the 5-6th week), this factor was vanishing 2 weeks after the delivery.