Immunosuppressive activity of culture supernatant from human decidual cells

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To assess the role of decidual cells in direct contact with paternal antigens in maintenance of pregnancy, immunosuppressive action of the culture suprenatant of decidual cells were investigated. Desidual cells were obtained from artificially aborted healthy pregnants on 8 to 10 weeks of gestational age. Dispersed cell suspensions were prepared by enzyme digestion method (collagenase 1mg/ml and DNase 0.05mg/ml). Desidual cells were enriched by Ficoll-Hypaque® density gradient, following dish adherent method. 1×106/ml of decidual cells were cultured for 48 hours in RPMI-1640 medium supplemented with 10% fetal calf serum, and the supernatants were harvested.The supernatant added at a concentration of 50% inhibited the reactions of human peripheral blood lymphocytes (PBL) by polyclonal mitotic activities of phytohemagglutinin (PHA), mixed lymphocytes culture (MLC), and killer T cell induction of PBL against alloantigen by 40%, 35% and 50%, respectively. To diminish the contamination of macrophages (spectific gravity: 1.045_??_1.075), discontinuous density gradient method using Percoll® was performed. Desidual cells were enriched over 90% in the fraction of specific gravity of 1.033 to 1.044 (fraction 2; Fr 2). The culture supernatant of the desidual cells (Fr 2) suppressed not only the mitotic activities of PHA to PBL by 29.6% at a concentration of 50%, but also the MLC by 40%, 19% and 8.75% at concentrations of 50%, 25%, 12.5%, respectively. The culture supernatant also suppressed the killer T cell induction by 53.3% and 13.8% at concentrations of 50% and 25%, respectively.To dissect the mechanism of immunosuppressive action of the culture supernatant, the suppressive effect of the supernatant on IL-2 production or IL-2 receptor expression of PBL stimulated by PHA, were investigated. The supernatant suppressed not only IL-2 production (0u/ml) but also IL-2 receptor expression (32.2%) compared with controlls (9.4u/ml, 48.1%). The supernatant also suppressed the immunoglobulin (IgG and IgA) production from PWM stimulated B cells separated from PBL. Those results indicate that desidual cells play a role in the maintenance of pregnancy by producing immunosuppressive substance which inhibit both T cell and B cell immunity to fetus.

Immunosuppressive activity of culture supernatant from human decidual cells

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