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本文应用被动皮肤过敏反应(PCA)检测大鼠淋巴器官中抗原特异性IgE和IgG 2a抗体形成细胞,以PCA滴度表示抗原特异性IgE和IgG 2a抗体的含量。用3 000条第三期钩虫幼虫(Nb)给Wistar鼠皮下注射,使其感染。6周后再以同样剂量的Nb再次感染,第二次感染的7—9天杀鼠取脾脏和肠系膜淋巴结(MLN),获取单个细胞悬液,计数细胞并用Hank′s液调节成适当浓度。B53抗DNP IgE或27—74.3抗dansyl IgE抗体杂交瘤细胞,以10~7个IgE杂交瘤细胞给小鼠腹腔内接种7～10天后收获腹腔渗出液内杂交瘤细胞,用Hank′s液调节成适当浓度。上述二种细胞悬液于-20℃冷冻,25℃融化,反复二次,4℃3 000rpm离心10分钟,获得细胞提
In this paper, passive anaphylaxis (PCA) was used to detect the antigen-specific IgE and IgG2a antibody-forming cells in the lymphoid organs of rats and the titer of PCA was used to express the antigen-specific IgE and IgG2a antibody. Wistar rats were injected subcutaneously with 3 000 third instar larvae (Nb) to infect them. Six weeks later, the cells were re-infected with the same dose of Nb. Spleens and mesenteric lymph nodes (MLN) were killed on the 7-9 days after the second infection. Single cell suspensions were obtained. Cells were counted and adjusted to the appropriate concentration with Hank’s solution. B53 anti-DNP IgE or 27-74.3 anti-dansyl IgE antibody hybridoma cells to 10 to 7 IgE hybridoma cells were intraperitoneally inoculated 7 to 10 days after the mice were harvested intraperitoneal exudate hybridoma cells with Hank’s solution Adjust to the appropriate concentration. The above two cell suspensions were frozen at -20 ° C and thawed at 25 ° C. The cells were repeatedly centrifuged at 3,000 rpm at 4 ° C for 10 minutes to obtain a cell extract