Human fibroblasts are permissive for porcine cytomegalovirus in vitro.
BACKGROUND
Xenotransplantation with pig organs is being considered to alleviate donor organ shortages; however, the risk of introducing porcine viruses into humans is heightened in this setting. The goal of this study was to determine the infectious potential of porcine cytomegalovirus (PCMV), a xenozoonotic virus of interest, in human fibroblasts in vitro.


METHODS
Confluent human cells were incubated with live PCMV, heat-killed PCMV, or medium alone. Infection was investigated by testing for viral-induced cytopathic effect, assaying viral transcription by nested RT-PCR and subsequent sequencing, and detecting viral protein expression by Western blotting. Plaque neutralization experiments were also performed.


RESULTS
Cells incubated with PCMV demonstrated significant cytopathic effect by 7 days postinfection, and reverse-transcriptase polymerase chain reaction sequencing identified PCMV DNA polymerase in these infected cells. In Western blots, monoclonal antibodies (mAbs) to human CMV glycoprotein B and pig serum presumed to contain anti-PCMV antibodies detected characteristic proteins in experimentally infected human cells and positive controls but not in negative controls. Furthermore, one of these mAbs and the pig serum neutralized PCMV infection in vitro.


CONCLUSIONS
These results are a first demonstration that PCMV can infect human fibroblasts in vitro.
Whitteker
Jennifer L
JL
Centre for Biologics Research, Biologics and Genetic Therapies Directorate, Health Canada, Ottawa ON, Canada.
Dudani
Anil K
AK
Tackaberry
Eilleen S
ES
D000818:Animals; D001483:Base Sequence; D015153:Blotting, Western; D002460:Cell Line; D002478:Cells, Cultured; D003587:Cytomegalovirus / Q000235:genetics / Q000276:immunology / Q000378:metabolism / Q000472:pathogenicity*; D003586:Cytomegalovirus Infections / Q000473:pathology / Q000635:transmission* / Q000821:virology; D003588:Cytopathogenic Effect, Viral; D005347:Fibroblasts / Q000473:pathology / Q000821:virology*; D052816:Foreskin / Q000821:virology; D006801:Humans; D008297:Male; D008969:Molecular Sequence Data; D009500:Neutralization Tests; D016377:Organ Transplantation / Q000009:adverse effects*; D012367:RNA, Viral / Q000378:metabolism; D020133:Reverse Transcriptase Polymerase Chain Reaction; D013552:Swine; D013997:Time Factors; D014183:Transplantation, Heterologous; D014759:Viral Envelope Proteins / Q000276:immunology; D010948:Viral Plaque Assay; D014764:Viral Proteins / Q000276:immunology / Q000378:metabolism
D012367:RNA, Viral; D014759:Viral Envelope Proteins; D014764:Viral Proteins; C040495:glycoprotein B, Simplexvirus