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Ishak, Ricardo; Linhares, Alexandre da Costa; Ishak, Marluísa de Oliveira Guimarães

doi:10.1590/s0036-46651989000200011

Cited by 7 publications

(2 citation statements)

References 2 publications

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“…Finally, attenuated or replication-defective HIV-1 may be used in research laboratories for the safety of laboratory workers (17)(18)(19)(20)(21). At least two cases of accidental, laboratoryacquired HIV-1 infection have been documented (21).…”

Section: Resultsmentioning

confidence: 99%

Derivation of a biologically contained replication system for human immunodeficiency virus type 1.

Chen

Boyle

Malim

et al. 1992

Proc. Natl. Acad. Sci. U.S.A.

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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 (vIJIBATat/Rev), which replicates only in a recombinant T-cell line expressing tat and rev (CEMTWRT), is described in this report. Infection of the CEM'r"T cell line with vIIIBATt/Rv permits the complete HIV-1 life cycle, including cytopathology, decreased expression of CD4, and production of viral structural proteins, to be bioiogically contained. Culture supernatants from infected CEM'r"T contain virus that is able to replicate only in uninfected CEMTART. To create a HIV-1 provirus functionally deficient in tat and rev, physically separate mutations in the first exon of tat and both exons of rev were made. The complementary Tat and Rev gene products were both expressed from a single bicistronic mRNA derived from a spliced cDNA. These reagents therefore provided a biologically contained system to study the replication of HIV-1 in culture. MATERIALS AND METHODSViruses. HIV-1 was generated as described by transfection of the proviral plasmid IIIB, a derivative of HXB-3 (4). Oligonucleotide-directed mutagenesis with a bacteriophage M13 system (Amersham) was used to introduce five point mutations into the coding region of Tat and Rev in pllUBTaT/Pev.Since Tat and Rev are encoded in different frames, mutations in the first exon of tat and rev result in a premature stop codon in tat while deleting the rev start codon. In the second exon of rev, mutations result in a premature stop codon. All mutations are shown in Table 1.The Tat-responsive chloramphenicol acetyltransferase (CAT) expression vector pLTR-CAT contains the HIV-1 long terminal repeat (LTR) fused to the CAT gene (4). The Rev-responsive CAT expression vector pDM128/CMV contains the simian virus 40 (SV40) origin of replication, the cytomegalovirus (CMV) immediate-early promoter, and the bacterial CAT gene inserted into an intron regulated by Rev. pBC12/CMV/CAT contains the CMV immediate-early promoter driving expression of the CAT gene (5) The publication costs of this article were defrayed in part by page charge payment. This article must therefore be hereby marked "advertisement" in accordance with 18 U.S.C. §1734 solely to indicate this fact.

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Section: Resultsmentioning

confidence: 99%

Derivation of a biologically contained replication system for human immunodeficiency virus type 1.

Chen

Boyle

Malim

et al. 1992

Proc. Natl. Acad. Sci. U.S.A.

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“…Outros pesquisadores também consideram os resíduos produzidos nas salas de emergência como infecciosos, principalmente pelo fato de que muito pouco se sabe sobre os pacientes dessas áreas (MEANEY & SCHEREMISIHOFF, 1989 GEDDES, 1986;GIBBS, 1990;OMS/CIE, 1990;GWYNTHER, 1990;ROBAZZI, 1991;RUTALA & WEBER, 1991;TRACHTMAN, 1991) Embora se encontrem poucos registros de doenças ocupacionais, comprovadamente causadas por agentes biológicos, os indivíduos que trabalham com agentes infecciosos ou suspeitos de conter patógenos, podem estar, de certa forma, expostos ao risco de infecção (SALVADOR, 1985;SIQUEIRA et al, 1989;ISHAK et al, 1989 (McCORMICK & MAKI, 1981;CDC, 1987;HORTON & ROGERS, 1990;BARNES, 1991).…”

Section: -Características Físico-químicasunclassified