摘要
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 3 5肽 ,并与等量弗氏佐剂混悬后免疫雌性 BALB/c小鼠。结果 :在 43 0 A固相自动肽合成仪上成功地合成了 3 5肽 ,并经 HPLC纯化、质谱鉴定证实其纯度达 95 %以上 ;新抗原免疫小鼠后获得的特异性抗血清可识别小鼠、大鼠及人睾丸组织蛋白提取物中相应分子量的蛋白质 ,且血清中特异性 Ig G抗体滴度最高达 1∶ 6 0 0 0 ,阴道粘膜冲洗液中特异性 Ig A抗体滴度最高达 1∶ 3 0 0。结论 :由含有两种特异性精子抗原、具有特异性氨基酸序列的新抗原所制备的抗血清可识别两种天然抗原 ,并在小鼠体内激发出较理想的特异性体液免疫 。
Objective: To develop a novel multivalent chimeric peptide vaccine for bisexual fertility regulation.Materials & Methods: On the basis of the amino acid sequence of the two peptides respectively selected from the mouse sperm/testis-specific proteins SP17 and Cyritestin,and one T cell epitope in bovine ribonuclease (RNase),a novel chimeric peptide consisting of 35 amino acid was designed and subsequently synthesized on the 430 A peptide synthesizer.After being emulified with the equivalence Freund's adjuvant,the peptide with 35 amino acid residues was used to immunize the female BALB/c mice to evaluate its immunity. Results: The peptide was successfully synthesized,after being purified in high performance liquid chromatography (HPLC),its purity reached 95%.The specific antisera collected from the immunogenity mice could identify the corresponding proteins of testis tissure of mice,rats and human.The highest specific IgG titer in serum was 1:6 000,while the IgA titer in the washing of vaginal mucous membrane was 1:300.Conclusion: The antibodies from the peptide with specific amino acid sequence can identify the original antigens,and stimulate powerful specific humoral immunity in mice.It provides a experimental bases for polyvalent contraceptive vaccine study.
出处
《生殖与避孕》
CAS
CSCD
北大核心
2002年第2期67-69,98,I001,共5页
Reproduction and Contraception
基金
国家自然科学基金资助项目
编号 :30 0 0 0 183
关键词
肽疫苗
避孕
精子
免疫
peptide vaccine,contraception,sperm immune
