Some morphological, growth, and genomic properties of human cells chronically infected with porcine endogenous retrovirus (PERV).
A major concern in using porcine organs for transplantation is the potential of transmission of porcine endogenous retrovirus (PERV). To investigate the long-term effects of PERV infection on human cells, human embryonic kidney cell line HEK-293 infected with PERV PK-15 was maintained for up to 72 passages and samples were harvested at intervals for use in morphological, growth, and genomic analyses. Morphology, DNA content/cell, and doubling time of uninfected and infected cells were similar. Restriction fragment length polymorphism (RFLP) analysis of PCR-amplified nearly full-length PERV genome showed no alterations in band pattern. RFLP analysis of the long terminal repeats (LTR) showed some changes in band pattern, but not in length. Southern blot analysis of genomic DNA of infected cells indicated random integration of PERV without structural alterations in proviral genome. Semi-quantitative PCR demonstrated a gradual increase of proviral load in the infected cells. Sequence analysis of the LTR region of PERV from infected cells indicated a relatively low rate (6.0 x 10(-4)/bp or about 2 x 10(-6)/bp/generation) of mutation. There were also indications of recombination of PERV strains A and B. Finally, PERV infection had no effect on transcription of human endogenous retrovirus-K (HERV-K) genes. Together, no significant effect attributable to PERV infection was evident on chronically PERV-infected HEK-293 cells.
Kuddus
Ruhul H
RH
Thomas E Starzl Transplantation Institute, Department of Surgery, University of Pittsburgh Medical Center, PA 15261, USA. kuddusru@uvsc.edu
Gandhi
Chandrashekhar R
CR
Rehman
Khaja K
KK
Guo
Fengli
F
Watkins
Simon C
SC
Valdivia
Luis A
LA
Fung
John J
JJ
D000818:Animals; D001483:Base Sequence; D018929:Cell Culture Techniques; D002460:Cell Line; D002999:Clone Cells; D020077:Endogenous Retroviruses / Q000235:genetics*; D006801:Humans; D007668:Kidney / Q000166:cytology* / Q000196:embryology / Q000821:virology*; D008969:Molecular Sequence Data; D012689:Sequence Homology, Nucleic Acid; D013552:Swine / Q000821:virology*; D020079:Terminal Repeat Sequences / Q000235:genetics