Detection of porcine endogenous retrovirus in cultures of freshly isolated porcine bone marrow cells.
Pigs are under consideration as possible sources of organs for xenotransplantation in humans. The induction of hematopoietic microchimerism through xenotransplantation of source animal hematopoietic cells has been suggested as a means to induce tolerance in potential recipients. Because all porcine cells contain genetic information for porcine endogenous retrovirus (PERV), coculture techniques, reverse transcriptase (RT) and reverse transcriptase-polymerase chain reaction assays were used to determine whether infectious PERV is released from fresh porcine bone marrow cells cultured in the presence or absence of porcine cytokines. Human embryonic kidney cell line, HEK-293 cells cocultured with porcine bone marrow cells were positive for PERV RNA but never became positive for viral RT activity, suggesting the PERV infection was not productive. In contrast, high levels of RT activity was detected in porcine ST-IOWA cells after coculture, demonstrating that these cells became productively infected. PERV was released from cultured porcine bone marrow cells without stimulation, and combinations of the porcine hematopoietic cytokines, interleukin-3, granulocyte macrophage-colony stimulating factor and stem cell factor had no additional effect on the infectivity or in vitro tropism of released PERV virions.
McIntyre
Maritza C
MC
Division of Cellular and Gene Therapies, Center for Biologics Evaluation and Research, Food and Drug Administration, 8800 Rockville Pike, Bethesda, MD 20892, USA.
Kannan
Bhanumathi
B
Solano-Aguilar
Gloria I
GI
Wilson
Carolyn A
CA
Bloom
Eda T
ET
McIntyre
Matiza C
MC
D000818:Animals; D001854:Bone Marrow Cells / Q000166:cytology* / Q000821:virology*; D002478:Cells, Cultured; D020077:Endogenous Retroviruses / Q000235:genetics / Q000302:isolation & purification*; D006801:Humans; D066298:In Vitro Techniques; D007668:Kidney / Q000166:cytology; D012367:RNA, Viral / Q000032:analysis; D012192:Retroviridae Infections / Q000635:transmission* / Q000821:virology; D013552:Swine; D014183:Transplantation, Heterologous
D012367:RNA, Viral