Screening pigs for xenotransplantation in China: investigation of porcine endogenous retrovirus in Diannan small-eared pigs.

ABSTRACT

Porcine endogenous retrovirus (PERV), which integrates as a provirus into the genome of pig cells, is an important biosafety issue in xenotransplantation. Screening and analyzing the presence and expression of PERV will provide essential parameters for assessing the biosafety of donor sources. In the present study, we investigated the prevalence of PERV in Diannan small-eared pigs, a unique closed colony that is distributed in southern Yunnan Province in southwestern China. PCR was performed to amplify env-A, env-B, env-C, pol, gag, and mtDNA in peripheral blood samples. The results revealed that PERV env-A, env-B, pol, and gag were detected in all individuals, but env-C was deficient in most pigs, suggesting that the main subtypes of PERVs in Diannan small-eared pigs are PERV-A and PERV-B. Furthermore, PERV pol and the porcine housekeeping gene GAPDH were detected by RT-PCR in all peripheral blood samples, indicating that PERV had transcriptional activity. Finally, the consensus sequences of PERV-A and PERV-B were amplified and digested with KpnI and MboI. Interestingly, a total of seven digestion patterns were obtained, which is less than that observed in other pig breeds. The PCR products were cloned into the pUCm-T vector and sequenced. The results showed that all of the inserts were highly homologous to either PERV-A or PERV-B, and the ratios of PERV-A and PERV-B were 21.1% and 78.9%, respectively. These data suggest that Diannan small-eared pigs may be a candidate donor source for xenotransplantation.