Bose R, Nagy E, Fleetham J, Pattinson H A, Berczi I, Yang J
Department of Obstetrics, Gynecology and Reproductive Sciences, University of Manitoba, Winnipeg, Canada.
Immunol Lett. 1993 Sep;38(1):47-54. doi: 10.1016/0165-2478(93)90117-k.
Balb/c mice were immunized with pre-implantation embryo-associated immunosuppressor factor (EASF) (purified from embryo growth media of in vitro fertilized human ova). Hybridoma clones were produced by fusing their spleen cells with NS1 and P3X653 myeloma cell lines. The presence of specific anti-EASF monoclonal antibodies (mAb) in the hybridoma culture supernatants were tested by enzyme-linked immunosorbent assay. A total of 15 hybridoma clones were selected, and their products were purified and characterized. Each mAb bound specifically to its antigen in a dose-dependent manner. The affinity-purified EASF from embryo growth media demonstrated immunosuppressive activity on Concanavalin A-induced lymphocytes and the presence of 14 kDa, 24 kDa and 37 kDa factors. No such activity or similar molecules were identified when control growth media were analyzed. This clearly demonstrates that these mAb are indeed EASF-specific and are able to recognize biologically active immunosuppressive components in embryo growth media. These mAbs are presently being tested for the development of EASF-specific assay system.
用植入前胚胎相关免疫抑制因子（EASF）（从体外受精人卵的胚胎生长培养基中纯化）免疫Balb/c小鼠。通过将其脾细胞与NS1和P3X653骨髓瘤细胞系融合产生杂交瘤克隆。通过酶联免疫吸附测定法检测杂交瘤培养上清液中特异性抗EASF单克隆抗体（mAb）的存在。总共选择了15个杂交瘤克隆，并对其产物进行了纯化和表征。每种mAb以剂量依赖的方式特异性结合其抗原。从胚胎生长培养基中亲和纯化的EASF对刀豆球蛋白A诱导的淋巴细胞表现出免疫抑制活性，并存在14 kDa、24 kDa和37 kDa的因子。分析对照生长培养基时未发现此类活性或类似分子。这清楚地表明这些mAb确实是EASF特异性的，并且能够识别胚胎生长培养基中的生物活性免疫抑制成分。目前正在测试这些mAb以开发EASF特异性检测系统。
