An in-vivo infectivity assay for cloned retroviruses lacking a susceptible cell culture.
The lymphoproliferative disease virus (LPDV) of turkeys is the retroviral agent of etiology of a rapidly developing, naturally occurring, lymphoproliferative process. Recently we have molecularly cloned the viral genome. The lack of a susceptible cell culture which can sustain LPDV replication hampered the analysis of the infectious capability of the cloned genome. Based on the efficient in-vivo replication of LPDV we have developed a sensitive in-vivo approach aimed at establishing the infectious capability of the cloned provirus. According to this approach, peripheral leukocytes withdrawn from 3-week-old turkeys were transfected with the cloned DNA and the transfected leukocytes were re-injected into the turkey from which they had been obtained. The injected leukocytes enabled the efficient expression of the viral genome and the release into the blood stream of LPDV virions, which thereafter could travel to their appropriate in-vivo target lymphoid cells and start multiple replication cycles, resulting in the development of a detectable viremia. The applicability of this in-vivo assay for other cloned viral genomes is discussed.
Department of Human Microbiology, Sackler School of Medicine, Tel-Aviv University, Israel.
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Gak
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Yaniv
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Ianconescu
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SR
Tronick
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Gazit
D000818:Animals; D003001:Cloning, Molecular; D004279:DNA, Viral / Q000302:isolation & purification*; D008232:Lymphoproliferative Disorders / Q000382:microbiology*; D008297:Male; D008828:Microbiological Techniques; D012194:RNA-Directed DNA Polymerase / Q000097:blood; D012190:Retroviridae / Q000235:genetics* / Q000472:pathogenicity; D012680:Sensitivity and Specificity; D014162:Transfection; D014422:Turkeys / Q000382:microbiology
D004279:DNA, Viral; D012194:RNA-Directed DNA Polymerase