几种重组腺相关病毒亚型载体对小鼠睾丸细胞感染和外源基因表达的特性研究  

Characterization of several recombinant adeno-associated virus subtypes on testicular cell infection and foreign gene expression in mice

作　　者：余欣耐 王斌驿 刘镕[1] YU Xinnai;WANG Binyi;LIU Rong(Taikang Medical College(School of Basic Medical Sciences),Wuhan University,Wuhan 430071,China)

机构地区：[1]武汉大学泰康医学院(基础医学院),武汉430071

出　　处：《数理医药学杂志》2024年第4期260-265,共6页Journal of Mathematical Medicine

基　　金：湖北省自然科学基金面上项目(2022CFB010)。

摘　　要：目的评估几种重组腺相关病毒载体rAAV1、rAAV2、rAAV2/8和rAAV6对小鼠睾丸细胞的感染特性。方法使用HEK293T细胞进行rAAV包装制备病毒颗粒,经生精小管内注射或睾丸间质注射给小鼠睾丸接种病毒,术后第7天用颈椎脱臼法处死小鼠,取睾丸制作冰冻切片,通过免疫荧光染色检测和评估重组腺相关病毒载体对睾丸细胞的感染情况和外源蛋白表达情况。结果rAAV1、rAAV2、rAAV6三种亚型均可感染支持细胞和生殖细胞,rAAV2对支持细胞感染能力更强,rAAV2/8在经生精小管接种病毒时特异地感染支持细胞,在经间质接种时特异性感染间质细胞。结论rAAV1、rAAV6在睾丸细胞感染中并未表现出明显的偏好性;rAAV2在可以感染生殖细胞和支持细胞的同时表现出对支持细胞的偏好性;rAAV2/8无法穿越血睾屏障,在经生精小管接种时特异性感染支持细胞,在经间质接种时特异性感染间质细胞。Objective To evaluate the infection characteristics of several recombinant adeno-associated virus(rAAV)subtypes including rAAV1,rAAV2,rAAV2/8,and rAAV6 on cells within the mouse testis.Methods HEK293T cells were used for rAAV packaging.The viruses were inoculated via intratubular injection or testicular interstitial injection to the mice.On the 7th day postinoculation,testes were collected to make frozen sections after mice were sacrificed by cervical dislocation,and immunofluorescence staining was performed to examine and evaluate the infection and foreign protein expression of rAAV vectors to testicular cells.Results rAAV1,rAAV2 and rAAV6 subtypes were capable of infecting both sertoli cells and germ cells,with rAAV2 exhibiting a relative stronger infection capability on sertoli cells.rAAV2/8 demonstrated an infection preference specifically for sertoli cells when inoculated via the seminiferous tubules,while it specifically infected leydig cells when inoculated via testicular interstitia.Conclusion rAAV1 and rAAV6 did not show specific infection preference in testicular cells.rAAV2 displayed a relative preference towards sertoli cells though it was able to infect both germ cells and sertoli cells.rAAV2/8 was unable to penetrate the blood-testis barrier,showing specific infection on sertoli cells when inoculated via seminiferous tubules and on leydig cells when inoculated via testicular interstitia.

关 键 词：腺相关病毒 睾丸 支持细胞 间质细胞 生殖细胞 外源基因递送 

分 类 号：R698.2[医药卫生—泌尿科学] R450[医药卫生—外科学]