Molecular and biological characterization of a replication competent human immunodeficiency type 2 (HIV-2) proviral clone.
We obtained complete genomic clones of human immunodeficiency virus type 2 (HIV-2) from the DNA of the neoplastic human cell line HUT 78 freshly infected with a HIV-2 isolate, strain SBL6669. The recombinant phage DNA was transfected into the lymphocytes of CD4-positive HUT 78 cell line to test the replication competence of the proviral DNA. One genomic clone, designated HIV-2SBL/ISY, yielded retroviral particles after a few weeks of culture of the transfected cells. The HIV-2SBL/ISY clone contained a complete provirus and cellular flanking sequence. We obtained the DNA sequence of the provirus and compared it with the published sequence of two other HIV-2 isolates. The degree of variability among HIV-2 isolates is comparable to that observed among African HIV-1 isolates sequenced to date. Immunologically, HIV-2SBL/ISY is similar to the parental virus (HIV-2SBL6669) but differs in the envelope transmembrane protein that is truncated (gp32-34) in the parental virus and not in HIV-2SBL/ISY (gp41). Both the parental and the cloned viruses are infectious and cytopathic for some human T-cell lines, induce syncytia, and infect a human macrophage cell line (U937) in vitro. The availability of a biologically active HIV-2 clone provides the means to study the role and interaction of HIV-2 genes in vitro as well as to assess the functional similarities among HIV-1 and HIV-2 genes. Since HIV-2SBL/ISY cloned virus infects fresh peripheral blood T cells from Rhesus macaques in vitro and infects the same animal in vivo, its use in animals may represent a model for functional study of viral genes in vivo as well as for development of experimental approaches to prevent and cure retroviral infection in humans.
Franchini
G
G
Laboratory of Tumor Cell Biology, National Cancer Institute, Bethesda, MD 20892.
Fargnoli
K A
KA
Giombini
F
F
Jagodzinski
L
L
De Rossi
A
A
Bosch
M
M
Biberfeld
G
G
Fenyo
E M
EM
Albert
J
J
Gallo
R C
RC
D015139:Blotting, Southern; D002460:Cell Line; D003001:Cloning, Molecular; D004279:DNA, Viral / Q000235:genetics* / Q000302:isolation & purification; D005455:Fluorescent Antibody Technique; D005814:Genes, Viral; D015498:HIV-2 / Q000235:genetics* / Q000302:isolation & purification / Q000502:physiology / Q000648:ultrastructure; D006801:Humans; D008854:Microscopy, Electron; D015183:Restriction Mapping; D013601:T-Lymphocytes; D014162:Transfection; D014779:Virus Replication
D004279:DNA, Viral