摘要
用亲和层析提取的肾综合征出血热病毒（ＨＦＲＳＶ）结构蛋白（５０ｋＤ和６７ｋＤ）为抗原，在体外免疫正常人外周血淋巴细胞（ＰＢＬ）。ＰＢＬ经亮氨酸甲基酯处理后，在含有ｓＰＷＭ－Ｔ、ＩＬ－２及不同浓度抗原的培养基中培养６ｄ，计数存活ＰＢＬ数量。结果表明，在上述体外免疫条件下存活ＰＢＬ的数量与抗原剂量呈正相关。将此体外免疫的ＰＢＬ与人－鼠杂交瘤细胞（Ｋ６Ｈ６／Ｂ５）融合，获得了３株分泌抗ＨＦＲＳＶ人单抗的杂交瘤细胞，表明在体外免疫条件下，正常人ＰＢＬ能够对ＨＦＲＳＶ结构蛋白发生特异性免疫应答。
T cell replacing factor(sPWM T) is obtained from pokeweed mitogen(PWM) stimulated T cells,during a short term culture.Isolated T cells by rosetting with sheep red blood cells were irradiated and subsequently cultured for 24 h in the presence of PWM and then remove the cells by centrifugation.The supernatnant were filtered and stored in a suitable aliquots at -20℃.PBL were obtained from healthy donors and treated with L leucyl L leucine methy(LeuLeu=OMe).The Leu Leu OMe treated PBL were in vitro immunized for 6 days with HFRSV protein in supplemented RPMI 1640,containing 50 μmol/L 2 mercaptoethanol,10% heat inactivated human serum,5 μ IL 2/ml,and 25% sPWM T.The results show that survival of PBL stimulated by HFRSV protein was significantly dependent on the dosage of the protein used during the six day in vitro immunization:the higher the dose of protein was used,the higher the viability of PBL.The cells in vitro immunized were then fused with K 6H 6/B 5 cells at a ratio 2∶1 and three positive clones were detected at 18 day.
出处
《中国免疫学杂志》
CAS
CSCD
北大核心
1997年第2期108-110,共3页
Chinese Journal of Immunology
