DERIVATION OF A BIOLOGICALLY CONTAINED REPLICATION SYSTEM FOR HUMAN-IMMUNODEFICIENCY-VIRUS TYPE-1

Human immunodeficiency virus type 1 (HIV-1) proviral mutants that lack viral regulatory genes are unable to replicate unless rescued by complementation in trans. Structurally intact virus can be produced by infecting recombinant cell lines expressing the deficient genes. A HIV-1 mutant functionally defective in tat and rev (vIIIB(DELTA-Tat/Rev)), which replicates only in a recombinant T-cell line expressing tat and rev (CEM(TART)), is described in this report. Infection of the CEM(TART) cell line with vIIIB(DELTA-Tat/Rev) permits the complete HIV-1 life cycle, including cytopathology, decreased expression of CD4, and production of viral structural proteins, to be biologically contained. Culture supernatants from infected CEM(TART) contain virus that is able to replicate only in uninfected CEM(TART). No reversion of vIIIB(DELTA-Tat/Rev) to wild-type HIV-1 was observed as measured either by sequencing proviral vIIIB(DELTA-Tat/Rev) or by detecting the ability of vIIIB(DELTA-Tat/Rev) to replicate in CEM or activated CD4-bearing T lymphocytes. Defective HIV-1 mutants produced by trans complementation of essential genes permit infection and analysis of defined genotypes on cellular function and phenotype. Authentic HIV-1 structural proteins and infected cells can be prepared in mass, and agents that interfere with the HIV-1 life cycle can be studied on a large scale with minimum risk of exposing workers to virulent HIV-1.