LUNAM University, Oniris, Nantes-Atlantic National College of Veterinary Medicine, Food Science and Engineering, Sanitary Security of Reproduction Biotechnology Unit, F-44307 Nantes, France.
Vet Res. 2012 Jan 25;43(1):5. doi: 10.1186/1297-9716-43-5.
The aim of this study was to determine, using immunofluorescence and in situ hybridization, whether CAEV is capable of infecting goat uterine epithelial cells in vivo. Five CAEV seropositive goats confirmed as infected using double nested polymerase chain reaction (dnPCR) on leucocytes and on vaginal secretions were used as CAEV positive goats. Five CAEV-free goats were used as controls. Samples from the uterine horn were prepared for dnPCR, in situ hybridization, and immunofluorescence. The results from dnPCR confirmed the presence of CAEV proviral DNA in the uterine horn samples of infected goats whereas no CAEV proviral DNA was detected in samples taken from the uninfected control goats. The in situ hybridization probe was complementary to part of the CAEV gag gene and confirmed the presence of CAEV nucleic acids in uterine samples. The positively staining cells were seen concentrated in the mucosa of the lamina propria of uterine sections. Finally, laser confocal analysis of double p28/cytokeratin immunolabelled transverse sections of CAEV infected goat uterus, demonstrated that the virus was localized in glandular and epithelial cells. This study clearly demonstrates that goat uterine epithelial cells are susceptible to CAEV infection in vivo. This finding could help to further our understanding of the epidemiology of CAEV, and in particular the possibility of vertical transmission.
本研究旨在通过免疫荧光和原位杂交技术，确定 CAEV 是否能够在体内感染山羊的子宫上皮细胞。使用白细胞和阴道分泌物的双重巢式聚合酶链反应 (dnPCR) 确认感染的 5 只 CAEV 血清阳性山羊被视为 CAEV 阳性山羊。5 只 CAEV 阴性山羊被用作对照。从子宫角制备样本进行 dnPCR、原位杂交和免疫荧光。dnPCR 的结果证实感染山羊的子宫角样本中存在 CAEV 前病毒 DNA，而未感染对照山羊的样本中未检测到 CAEV 前病毒 DNA。原位杂交探针与 CAEV gag 基因的一部分互补，证实了子宫样本中存在 CAEV 核酸。阳性染色细胞集中在子宫切片固有膜的黏膜中。最后，对 CAEV 感染山羊子宫的双 p28/细胞角蛋白免疫标记横切片进行激光共聚焦分析，表明病毒定位于腺上皮细胞和上皮细胞。本研究清楚地表明，山羊的子宫上皮细胞容易受到 CAEV 的体内感染。这一发现有助于进一步了解 CAEV 的流行病学，特别是垂直传播的可能性。
