Immortalization of caprine fibroblasts permissive for replication of small ruminant lentiviruses.
OBJECTIVE
To establish immortalized caprine fibroblastic cell lines permissive for replication of small ruminant lentiviruses.


METHODS
Carpal synovial membrane explants collected aseptically from a surgically delivered fetus of a lentivirus-seronegative goat.


METHODS
Immortalization of goat embryonic fibroblasts was performed by DNA transfection with plasmids coding for simian virus 40 large T antigen. The generated cell lines were phenotypically characterized. Cytogenetics, growth pattern, and sensitivity to viral infection were studied.


RESULTS
3 cell lines, designated TIGEF, mMTSV-54, and mMTSV-93, were generated. They had a more rapid doubling time than did fibroblasts from which they were derived, and retained morphologic and phenotypic fibroblastic characteristics. They were immortalized but not transformed because tumor formation was not promoted after their s.c. injection into athymic nude mice. The 3 cell lines were susceptible to caprine arthritis-encephalitis virus and visna-maedi virus infections, and supported a complete virus replication cycle.


CONCLUSIONS
Cultured caprine fibroblastic cells were immortalized, using simian virus 40 large T antigen. The TIGEF, mMTSV-54, and mMTSV-93 immortalized cell lines were permissive to in vitro small ruminant lentivirus replication.


CONCLUSIONS
Because lentivirus detection, as well as detailed studies of host-lentivirus interactions, are hampered by differences in viral susceptibility of each primary culture, permanent cell lines are essential tools for such analysis.
Da Silva Teixeira
M F
MF
Laboratoire associé de recherches sur les lentivirus des petits ruminants: INRA and Ecole Nationale vétérinaire de Lyon, France.
Lambert
V
V
Mselli-Lakahl
L
L
Chettab
A
A
Chebloune
Y
Y
Mornex
J F
JF
D000818:Animals; D000952:Antigens, Polyomavirus Transforming / Q000235:genetics; D016088:Arthritis-Encephalitis Virus, Caprine / Q000235:genetics / Q000302:isolation & purification / Q000502:physiology; D002455:Cell Division / Q000502:physiology; D002460:Cell Line; D018076:DNA, Complementary / Q000235:genetics; D004279:DNA, Viral / Q000235:genetics; D005347:Fibroblasts / Q000166:cytology* / Q000821:virology*; D006041:Goats; D017972:Lentiviruses, Ovine-Caprine / Q000235:genetics / Q000302:isolation & purification / Q000502:physiology*; D051379:Mice; D008819:Mice, Nude; D010641:Phenotype; D010957:Plasmids; D011003:Ploidies; D016133:Polymerase Chain Reaction / Q000662:veterinary; D012680:Sensitivity and Specificity; D012756:Sheep; D013583:Synovial Membrane / Q000166:cytology / Q000196:embryology; D014162:Transfection; D014779:Virus Replication; D014790:Visna-maedi virus / Q000235:genetics / Q000302:isolation & purification / Q000502:physiology
D000952:Antigens, Polyomavirus Transforming; D018076:DNA, Complementary; D004279:DNA, Viral