编码精子赤道段蛋白的质粒DNA免疫影响小鼠生育功能  

Antifertility effect in mice after vaccination with plasmid DNA encoding mouse sperm equatorial segment protein

作　　者：吕正梅[1,2] 王旻[1,3] 徐晨[1,3] 

机构地区：[1]上海交通大学医学院组织学胚胎学教研室,上海200025 [2]安徽医科大学组织学胚胎学教研室,合肥230032 [3]上海市生殖医学重点实验室,上海200025

出　　处：《解剖学报》2011年第1期129-132,共4页Acta Anatomica Sinica

基　　金：国家自然科学基金资助项目(81070526)

摘　　要：目的探讨编码小鼠精子赤道段蛋白(mESP)的DNA疫苗的免疫避孕功能。方法构建pcDNA3.1/mESP真核表达质粒,表达mESP蛋白N-端片段。将纯化后的质粒直接肌肉注射免疫8只BALB/c雌性小鼠,以空载质粒pcDNA3.1免疫作为对照组。通过RT-PCR检测重组质粒在小鼠体内的表达情况。通过ELISA和双重免疫荧光技术,检测免疫后小鼠体内mESP抗体反应。通过交配实验研究DNA免疫对小鼠生育功能的影响。结果接种重组质粒小鼠的肌肉呈现mESP cDNA条带阳性,说明重组质粒在体内表达。DNA免疫小鼠体内产生了特异性的mESP抗体。DNA免疫小鼠产仔率无改变,平均产仔数低于对照组(P<0.05)。结论 DNA疫苗用于免疫避孕需进一步提高免疫效果。Objective To determine the feasibility of pcDNA3. 1/mESP as a potential immunocontraceptive antigen. Methods Eukaryotic expression plasmid pcDNA3. 1/mESP, encoding N-terminal of mouse sperm equatorial segment protein （mESP）, was constructed. Two groups of mice （n = 8/group） were inoculated with plasmids of pcDNA3.1/mESP and peDNA3. 1 respectively. RT-PCR, ELISA and double immunofluorescence assay were performed to observe peDNA3.1/mESP expression and immune response in the inoculated mice. Mouse mating test was employed to study fertility of the experimental and control group. Results mESP cDNA positive band was detected in muscle from mice immunized with peDNA3. 1/mESP, which identified the expression of the recombinant plasmid. The pcDNA3. 1/mESP induced specific antibody against mESP in the experimental mice. Significant difference of mean litter size between experimental and control groups was observed, while the rate of fertility did not decrease. Conclusion This study indicates the need to strengthen DNA vaccines in order to obtain effective immunocontraception.

关 键 词：精子赤道段蛋白 DNA疫苗 免疫避孕 双重免疫荧光技术 反转录一聚合酶链式反应 小鼠 

分 类 号：R321.1[医药卫生—人体解剖和组织胚胎学]