RESUMO
The small ruminant lentiviruses, namely caprine arthritis encephalitis virus (CAEV) and Maedi Visna virus (MVV) are grown currently in secondary synovial membrane cells. Primary and secondary cell cultures are sometimes difficult to obtain and support a low number of passages and, therefore, permissive cell lines are needed. A transformed cell line was obtained by transfection of ovine synovial membrane secondary cell culture with a plasmid containing the SV40 large T antigen gene. The transformed cell culture described in this paper showed a higher growth rate and a more homogenous population of fibroblast-like cells when compared to the original ovine synovial membrane secondary cell cultures. Karyotype analysis has indicated the induction of many random chromosome changes, leading to a decrease in chromosome number. The SV40 DNA was detected in the nucleus and in the cytoplasm of transformed cells. The putative expression of large T antigen was presumed by the detection of the corresponding mRNA by PCR. Finally, the transformed ovine synovial membrane cells were shown to be permissive to small ruminant lentiviruses, and these are suggested as a cell line for in vitro isolation and propagation of these viruses.
Assuntos
Antígenos Transformantes de Poliomavirus/metabolismo , Transformação Celular Viral , Fibroblastos/virologia , Vírus 40 dos Símios/metabolismo , Membrana Sinovial/virologia , Animais , Antígenos Transformantes de Poliomavirus/genética , Vírus da Artrite-Encefalite Caprina/fisiologia , Linhagem Celular , Linhagem Celular Transformada , DNA Viral/análise , DNA Viral/genética , Fibroblastos/fisiologia , Cariotipagem , RNA Mensageiro/análise , RNA Mensageiro/genética , Ovinos , Vírus 40 dos Símios/genética , Vírus 40 dos Símios/fisiologia , Membrana Sinovial/citologia , Transfecção , Virologia/métodos , Vírus Visna-Maedi/fisiologiaRESUMO
This study demonstrated the presence of viral particles suggestive of arthritis-encephalitis virus through the clinic, serologic, pathologic, immunohistochemic and ultrastructural studies in dairy goats. In the postmortem studies the tissues used were: synovial membrane, lungs and mammary gland from goats with clinical signs characteristic of the disease and positive for serum antibodies to the virus.