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J. Virol., Dec 1997, 9124-9133, Vol 71, No. 12
Copyright © 1997, American Society for Microbiology

The superantigen-homologous viral immediate-early gene ie14/vsag in herpesvirus saimiri-transformed human T cells

A Knappe, C Hiller, M Thurau, S Wittmann, H Hofmann, B Fleckenstein and H Fickenscher
Institut fur Klinische und Molekulare Virologie, Friedrich-Alexander- Universitat Erlangen-Nurnberg, Erlangen, Germany.

Herpesvirus saimiri C488 transforms human T lymphocytes to stable growth in culture. The growth-transformed human T cells harbor the viral genome in a nonintegrated episomal form without production of virus particles. In these cells, virus gene expression was previously found to be confined to the transforming genes stpC and tip. In order to analyze virus gene expression in more detail, we applied a subtractive hybridization technique and compared stimulated virus- transformed cells with uninfected parental T cells of the same donor. A number of known T-cell activation genes were isolated. Viral stpC/tip cDNAs were enriched after subtraction. In addition, the viral immediate- early, superantigen-homologous gene ie14/vsag was represented by numerous cDNA clones that comprised the entire spliced transcript. Whereas a weak basal expression of ie14/vsag was detected by reverse transcription-PCR only, the phorbol ester-induced transcripts were readily shown by Northern blotting. ie14/vsag, which before had been classified as a major immediate-early gene of herpesvirus saimiri, is localized within a highly conserved region with extensive homologies to the cellular genome. Mutant viruses without the ie14/vsag gene are replication competent and fully capable of transforming human and marmoset T cells. Since ie14/vsag is transiently expressed after stimulation, it may increase T-cell proliferation in an activation- dependent and superantigen-like but apparently Vbeta-independent way.

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