Long-term follow-up failed to detect in vitro transmission of full-length porcine endogenous retroviruses from specific pathogen-free pig islets to human cells
Long-term follow-up failed to detect in vitro transmission of full-length porcine endogenous retroviruses from specific pathogen-free pig islets to human cells
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DOI：
10.1007/s001250100010
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发表时间：
2001-11-01
期刊：
影响因子：
8.2
通讯作者：
Saï, P
中科院分区：
文献类型：
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作者：
Clémenceau, B;Jégou, D;Saï, P
目的/假设。无特定病原体（SPF）猪的胰岛可以预防传统人兽共患病的传播。而不是内源性逆转录病毒（PERV），从猪到糖尿病患者。我们以前报道过，胰腺中PERV mRNA的表达最低的猪组织移植。本研究旨在确定来自猪胰岛的PERV是否在共孵育期间感染人细胞。将人细胞（包括高度PERV敏感的293细胞）与SPF猪胰岛细胞在设计为增加接触的条件下孵育（胰岛与人细胞的高比率，延长的共培养时间）。反复接触）。使用产生PK 15和G2逆转录病毒的猪细胞代替胰岛细胞作为“阳性感染对照”。通过PCR或长PCR以及RT-PCR或长RT-PCR在细胞提取物和上清液上监测人细胞的感染。检测PERV DNA和mRNA。分别用PERT法检测逆转录酶活性。尽管猪胰岛细胞中存在所有PERV序列，包括全长插入片段。在18周的随访期间，在与猪胰岛细胞一起孵育的任何人细胞系或血液单核细胞中均未检测到gag、pol和3种env亚型的DNA或RNA。在上清液中未检测到PERV序列或RT活性。即使当猪胰岛与人细胞的比例增加到100：1，共培养时间延长到5天并进行两次连续共孵育时，PERV信号也是阴性的。相比之下，在与PK 15或G2细胞共孵育的所有人细胞中检测到所有PERV DNA和mRNA。根据人细胞类型，获得生产性或非生产性感染：检测或不检测上清液中的全长PERV RNA和RT活性;以及通过PERV感染的人钟向先前未暴露的人细胞传递或不传递PERV序列。一些人细胞没有被PK 15细胞有效感染，但在与PERV感染的293细胞共孵育后变得有效感染。SPF猪胰岛细胞，即使有PERV插入物和转录物。在共孵育过程中将PERV传播到人类细胞的可能性非常小。人细胞对PERV稳定和生产性感染的敏感性取决于细胞类型。观察到PERV的人类适应性。
Aims/hypothesis. Islets from specific pathogen-free (SPF) pigs could prevent the transmission of conventional zoonosis. but not endogenous retroviruses (PERV), from pigs to diabetic patients. We previously reported that the pancreas showed the lowest expression of PERV mRNA among pig tissues intended for grafting. This study aimed to determine whether PERV from pig islets infect human cells during co-incubation.Methods. Human cells (including highly PERV-sensitive 293 cells) were incubated with SPF pig islet cells under conditions designed to increase contact (a high islet to human cell ratio, extended period of coculture. and repeated contacts). PK15 and G2 retrovirus-producing pig cells were used in place of islet cells as "positive infection controls". Infection of human cells was monitored on cellular extracts and supernatants by PCR or long PCR, and RT-PCR or long RT-PCR. to detect PERV DNA and mRNA. respectively. Reverse- transcriptase activity was monitored by PERT.Results. Despite the presence of all PERV sequences in pig islet cells, including full-length inserts. no DNA or RNA for gag, pol, and the 3 env sub-types were detected in any human cell line or blood mono- nuclear cells incubated with pig islet cells, during an 18-week follow-up period. No PERV sequences or RT activity were detected in supernatants. PERV signals were negative even when the pig islet to human cell ratio was increased to 100:1, the time of co-culture was extended to 5 days and two sequential co-incubations were done. By contrast, all PERV DNA and mRNA were detected in all human cells co-incubated with PK15 or G2 cells. Depending on human cell types, productive or non-productive infections were obtained: full-length PERV RNA and RT activity in supernatants were detected or not; and PERV sequences to previously unexposed human cells by PERV-infected human bells were transmitted or not. Some human cells were not productively infected by PK15 cells but became productively infected after co-incubation with PERV-infected 293 cells.Conclusion/interpretation. SPF pig islet cells, even with PERV inserts and transcripts. have very little probability of transmitting PERV to human cells during co-incubation. The sensitivity of human cells to stable and productive infection by PERV depends on the cell type. Human adaptation of PERV was observed.