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Virus-free survival and down-regulation of CD4 in C8166 cells infected with human immunodeficiency virus type 1 at low density

¹ Centre of Virology, S. Camillo Hospital, USL RM 10, Circonvallazione Gianicolense 85, 00152 Rome
and² Laboratory of Cell Biology, Istituto Superiore di Sanità, Viale Regina Elena 299, 00161 Rome, Italy

Compared with other T cell lines, C8166 lymphocytes are particularly susceptible to human immunodeficiency virus (HIV) infection and the outcome is invariably cell death. The results reported in this study demonstrate that the virus-induced cytolysis is strongly dependent on the initial cell density of C8166 cultures. Cultures diluted to 50 to 500 cells/ml almost completely maintained their cell duplication rate and released infectious virus into the medium. HIV infection of diluted C8166 cells is a simple and easily reproducible procedure for obtaining persistently infected cultures. These cultures contained genomic and extragenomic HIV DNA, the latter being assayed by PCR for two-long terminal repeat circular forms. The status of persistent infection disappeared within 2 months. The recovery is due to the replacement of CD4 down-regulated infected cells by overgrowing uninfected cell variants, which are transcriptionally inactive for CD4. The mechanisms underlying the emergence of these variants in persistently infected cultures are considered.

Received 20 April 1993; accepted 27 July 1993.

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