생쥐의 영양아층세포 배양액으로부터 면역조절물질의 순화 및 세포화학적 연구 = Purification and Cytochemical Study of Immune Regulation Substance in Murine Trophoblast Culture Supernatant

다국어 초록 (Multilingual Abstract)

The present studies have been, therefore, attempted in order to elucidate the mechanism by which protection of the feto-placental allograft, between the C3H/HeJ female mouse and DBA/2 male mouse occurred. For this purpose, immune suppressive substance from trophoblast cell culture supernatant was purified and characterized. And immunocytochemical study of the purified substance in placenta were performed. The immune suppressive substance was purified from the culture supernatant of trophoblast cells by means of ultra centrifugation, Sephacryl S-200 gel filtration chromatography and DEAE-trisacryl M anion exchange chromatography. The purified substance suppressed natural killer cell cytotoxicity, antibody dependent cell cytotoxicity, lymphokine activated killer cell cytotoxicity, and also suppressed the cytotoxic T cell generation and interleukin-2 production by mixed lymphocytes reactions. The immune suppressive effect of the purified substance was apparently abolished and activated by the treatment of excessive interleukin-2. The purification yield of the purified substance was 10.7%, and the molecular weight of which was turned out to be 15KD by SDS PAGE. Quantitating prostaglandin EZ in the purified subtance, 0.64 pg/ml of it was found to be included in the 10 pg/ml of protein. The purified substance was observed in the cytoplasm of syncytiotrophoblast cells in the placenta with anti serum obtained from a rabbit by means of the immunocytochemical staining method. It is, therefore, concluded that during the pregnancy, the purified immune suppressive substance from the trophoblast culture supernatant in this study synthesized in syncytiotrophoblast might regulate the activity of maternal immune effector cells against fetal cells by modulating activity of interleukin-2, so contribute to protection of the fetus as an allograft from the maternal immunological attack.