LUNAM University, Oniris, Nantes-Atlantic National College of Veterinary Medicine, Food Science and Engineering, Sanitary Security of Reproduction Biotechnology Unit, Nantes F-44307, France.
Comp Immunol Microbiol Infect Dis. 2013 Jul;36(4):397-404. doi: 10.1016/j.cimid.2013.02.004. Epub 2013 Apr 25.
The transmission of CAEV from male goats has not been well studied and the target cells that support viral replication are not well characterized. Epididymal epithelial cells (EECs) are important and play a key role in the fertility and motility of spermatozoa. During their transit, spermatozoa incorporate several EEC-produced proteins into their plasma membranes to stabilize them and prevent premature acrosomal reaction. This intimate interaction between spermatozoa and EECs may increase the likelihood of the infection of semen with CAEV if epididymal tissue is productively infected and sheds the virus into the duct. The aim of this study was to examine whether goat EECs are susceptible to CAEV infection in tissue culture. Cells were isolated from epididymides obtained from goats that were sampled from a certified-CAEV-free herd. Cultured cells were then inoculated with a molecularly-cloned isolate of CAEV (CAEV-pBSCA). Inoculated cells developed cytopathic effects (CPE), showing numerous multinucleated giant cells (MGC) in cell-culture monolayers. Expression of CAEV proteins was detected by immunofluorescence using an anti-p28, Gag-specific antibody. The culture medium of inoculated cells was shown to contain high titers (10(6) tissue culture infectious doses 50 per ml (TCID50/ml)) of infectious, cytopathic virus when assayed using indicator goat synovial membrane (GSM) cells. Our findings clearly demonstrate that cells of the buck genital tract are targets of CAEV and are thus a potential reservoir that sheds infectious CAEV into the semen of infected animals. These data suggest the use of sperm from CAEV-free goat males for artificial insemination in genetic selection programs to minimize CAEV dissemination.
CAEV 从雄性山羊传播的情况尚未得到很好的研究，支持病毒复制的靶细胞也没有很好地描述。附睾上皮细胞（EEC）是重要的，在精子的生育和活力中起着关键作用。在它们的运输过程中，精子将几种 EEC 产生的蛋白质整合到它们的质膜中，以稳定它们并防止过早的顶体反应。如果附睾组织受到感染并将病毒释放到管中，精子与 EEC 之间的这种密切相互作用可能会增加精液感染 CAEV 的可能性。本研究旨在研究组织培养中的山羊 EEC 是否容易感染 CAEV。细胞从从经过 CAEV 认证的无病群中采样的山羊的附睾中分离出来。然后将培养的细胞用分子克隆的 CAEV 分离株（CAEV-pBSCA）接种。接种的细胞出现细胞病变效应（CPE），在细胞培养单层中显示出许多多核巨细胞（MGC）。通过使用抗 p28、Gag 特异性抗体的免疫荧光检测到 CAEV 蛋白的表达。接种细胞的培养基在使用指示山羊滑膜（GSM）细胞测定时显示出高滴度（10(6)组织培养感染剂量 50 每毫升（TCID50/ml））的传染性、致病变病毒。我们的研究结果清楚地表明，公畜生殖道的细胞是 CAEV 的靶细胞，因此是一种潜在的储库，可将感染性 CAEV 释放到感染动物的精液中。这些数据表明，在遗传选择计划中，应使用来自无 CAEV 山羊雄性的精子进行人工授精，以最大程度地减少 CAEV 的传播。
