1College
of Animal Science and Technology, Guangxi University, Nanning
530004,
Guangxi,
China 2Guangxi
Veterinary Research Institute, Nanning,
530001, Guangxi,
China
*Corresponding author:
wu-jm20@163.com
Abstract
Bama miniature pig (BM) as the inbreed pigs is
considered to be a potential donor for xenotransplantation. However, porcine
endogenous retrovirus (PERV) integrated in pig’s genome, is likely to infect
human cells in vitro. PERV infection from BM remains unknown. This study aimed
to investigate PERV infection in BM in comparison with that of PK15 cells and
further to identify the integration sites of PERV from BM (PERV-BM) in human
genome. Two full-length PERV-BMs were cloned from 15 isolates of PERV from Bama
miniature pigs and sequence analysis indicated that PERV-BM had limited
infection to human cells. Subsequently, PBMC from the BM and PK15 cells were
co-cultured with HEK293 cells, respectively. Using nested RT-PCR assay and TEM
observation, one PERV-BM was observed to infect HEK293 cells with lower
transcriptional level than PERV from PK15 cells (PERV-PK) did, while the others
showed no sign of transmission of PERV to human cells. Furthermore, 4 and 157
integration sites were identified from PERV-BM-infected HEK293 cells
(HEK293-PERV-BM) and PERV-PK-infected HEK293 cells (HEK293-PERV-PK),
respectively, which displayed variations of PERV integration. For further
verification of the PERV-BM integration, the expression level of
cyclin D1,
c-myc, p53 and
p16 genes of HEK293-PERV-BM decreased
significantly while the CDK4 genes
were upregulated dramatically in comparison with those of the HEK293-PERV-PK. It
is the first time that PERV-BM has been confirmed to be low infection to human
cells and highly integration into the PK15 cells. Therefore, BM is a potential xenograft
donor due to less PERV transmission to human beings.
To Cite This Article:
Ma L, Lin J, Tang H, Bai A, Chen F, Qin S, Liu J,
Qin S and Wu J, 2020. Characterization of
PERV from Bama miniature pigs in human cells. Pak Vet J, 40(1): 37-42. http://dx.doi.org/10.29261/pakvetj/2019.094