Simian immunodeficiency viruses have been isolated from four species of monkey, the 'captive' macaque and mangabey and the 'feral' African green monkey and mandrill. While none of these viruses is a replica of HIV-1, the macaque and mangabey viruses represent correct genetic models for HIV-2, possessing exactly the same complement of genes. Recently a lentivirus has been identified in two wild chimpanzees (Pan troglodytes troglodytes) in Gabon, west equatorial Africa, and isolated from one of them. This virus is referred to as SIVCPZ. Sera from these animals cross reacted with all the HIV-1 proteins including the envelope glycoproteins. Here, we describe the molecular cloning and sequencing of an infectious proviral clone of SIVCPZ. The overall genetic organization was the same as that of HIV-1, but phylogenetic analysis revealed that the sequence was more divergent than any HIV-1 sequence reported so far. The vpu gene product, found only in the type 1 viruses, was particularly different (64% divergent to HIV-1BRU) suggesting that the SIVCPZ represents a distinct subtype. These findings indicate that there is a larger pool of simian lentiviruses than previously suspected and revives debate as to the origins of HIV-1.

A series of triterpene compounds characterized by a stringent structure-activity relationship were identified as potent and selective inhibitors of human immunodeficiency virus type 1 (HIV-1) replication. Currently studied betulinic derivatives have 50% inhibitory concentrations (ICss) agaeHIV-1 strain III/LAI in the 10 nM range in several cellular infection assays but are inactive against HIV-2. These compounds did not significantly inhibit the in vitro activities of several purified HIV-l enzymes. Rather, they ap ed to block virus infection at a postbinding, envelope-dependent step involved in the fusion of the virus to the cell membrane. CCID5o) and cultured for 5 days. Cell viability was assessed by the spectrophotometric 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium-based assay (9). Mock-infected cultures were carried out in parallel to determine the degree of cytotoxicity of the compounds. p24 core antigen in culture supernatants was determined by ELISA (NEK060B kit from New England Nuclear). RT activity was measured by a poly(rA) 3H SPA scintillation proximity assay (NK9020 from Amersham). The antiviral assay using MT4 cells was as described (10). The effect of compounds on the kinetics of viral production in human peripheral blood mononuclear cells (PBMCs) was also assayed. PBMCs from seronegative donors were isolated by Ficoll/Hypaque (Pharmacia) and stimulated for 3 days with phytohemagglutinin (2.5 pg/ml; Difco). Cells were then washed three times with medium, pelleted, and incubated with 50-100 CCID50 of virus stock per ml. After 1 hr at room temperature, cells were resuspended in RPMI 1640 medium containing 10%6 fetal bovine serum, 10%6 T-cell growth factors (Lymphocult, Biotest Diagnostics, Danville, NJ), and anti-interferon a neutralizing antibody (Bayer, Wuppertal, F.R.G.) at 80 units/ml. Cells were plated at 106 per ml in six-well culture plates (Costar) in the presence of the compound to be tested. At 3, 7, 10, and 14 days after infection, cultures were harvested and viable cells were examined by trypan blue exclusion. Aliquots of supernatants were frozen at -700C until tested for p24 antigen, and cells were resuspended at the same density in complete fresh medium with the same concentration of the tested drug.Determination of Cytoplasmic Proviral DNA. Shortly after infection, the proviral DNA obtained from a selective extraction was quantified by PCR amplification. H9 cells were preincubated in the presence or absence of the drug to be tested for 1 hr, infected with HIV-1 (LAI) at 500 CCIDso/ml for 1 hr, and grown in culture medium for another hour. Cells were then washed intensively, and cytoplasmic proviral DNA was extracted by a digitonin/proteinase K/RNase Abbreviations: AZT, 3'-azido-3'-deoxythymidine; HIV, human immunodeficiency virus; PBMC, peripheral blood mononuclear cell; sCD4, soluble CD4; RT, reverse transcriptase. tTo whom reprint requests should be addressed. 3564The publication costs of this article were defrayed in part by page charge payment. This article mus...

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A novel series of omega-aminoalkanoic acid derivatives of betulinic acid were synthesized and evaluated for their activity against human immunodeficiency virus (HIV). The anti-HIV-1 activity of several members of this new series was found to be in the nanomolar range in CEM 4 and MT-4 cell cultures. The optimization of the omega-aminoalkanoic acid side chain is described. The presence of an amide function within the side chain was found important for optimal activity. RPR 103611 (14g), a statine derivative, was found to be inactive against HIV-1 protease, reverse transcriptase, and integrase as well as on gp120/CD4 binding. "Time of addition" experiments suggested interaction with an early step of HIV-1 replication. As syncytium formation, but not virus-cell binding, seems to be affected, betulinic acid derivatives are assumed to interact with the postbinding virus-cell fusion process.