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目的 开发可供两性使用的多价合成嵌合肽疫苗。方法 选择小鼠精子 /睾丸特异性抗原SP1 7、Cyritestin中特异性氨基酸序列与牛核糖核酸酶非限制性T细胞表位作为骨架 ,按一定连接方式设计、合成新抗原 35肽 ,并与弗氏佐剂混悬后免疫雌性BALB/C小鼠。结果 在 430A固相自动肽合成仪上成功地合成 35肽 ,并经HPLC纯化、质谱鉴定证实其纯度达 95 %以上 ;免疫后小鼠血清特异性IgG抗体滴度最高达 1∶6 0 0 0 ,阴道粘膜冲洗液中特异性IgA抗体滴度最高达 1∶30 0 ;小鼠脾脏淋巴细胞增殖率达 50 %左右 ,淋巴细胞分泌的INF γ和IL 4分别为 60、1 0 8μg/ml;实验组BALB/C雌鼠妊娠率平均降至 60 % ,每胎产仔数平均降低 58%～71 %。结论 含有两种特异性精子抗原中特异性氨基酸序列的新抗原可激发小鼠产生较理想的特异性体液免疫和粘膜免疫 ,并使小鼠受孕率下降 ,为研制适宜于两性的多价抗生育疫苗提供了实验基础
Objective To develop a multivalent synthetic chimeric peptide vaccine for both sexes. Methods The specific amino acid sequence of mouse sperm / testis-specific antigen SP1 7, Cyritestin and bovine ribonuclease non-restricted T cell epitopes were selected as the backbone and designed according to a certain connection. The new peptide 35 peptide was synthesized and compared with Freund’s Female Balb / C mice were immunized after adjuvant suspension. Results The 35 peptide was successfully synthesized on a 430A solid-phase automatic peptide synthesizer and purified by HPLC. The purity of the 35 peptide was confirmed to be over 95% by mass spectrometry. The titer of serum-specific IgG antibody after immunization was up to 1: 600 , The specific IgA antibody titers in vaginal mucosal washings were up to 1:30 0; the proliferation rate of mouse spleen lymphocytes was about 50%; the levels of INF γ and IL 4 secreted by lymphocytes were 60 and 108 μg / ml; In the experimental group, the average pregnancy rate of BALB / C female mice dropped to 60% and the average number of litter size per litter decreased by 58% -71%. Conclusions The new antigen containing the specific amino acid sequence of the two specific sperm antigens can stimulate mice to produce more ideal specific humoral and mucosal immunity and reduce the pregnancy rate of mice. In order to develop multivalent anti-fertility suitable for both sexes Vaccines provide the experimental basis