Acquired immunodeficiency syndrome (AIDS) is a result of replication of the human immunodeficiency virus type 1 (HIV-1) predominantly in CD4؉ T lymphocytes and macrophages. However, most of these cells in vivo are immunologically quiescent, a condition restricting HIV-1 replication. Vpr is an HIV-1 virion protein suspected to enhance HIV-1 replication in vivo. We demonstrate in this report that Vpr specifically activates HIV-1 long terminal repeat (LTR)-directed transcription. This effect is most pronounced on a minimal promoter from HIV-1 LTR containing the TATA box and binding motifs for the ubiquitous cellular transcription factor Sp1. Evidence is presented that Vpr interacts with Sp1 when Sp1 is bound to the Sp1 motifs within the HIV-1 LTR. Both Vpr-Sp1 interaction and Vpr trans-activation require a central Leu/Ile-rich domain in Vpr. Our findings suggest that Vpr trans-activation through Sp1 is most critical for the immediate early transcription of HIV-1 when other positive regulators, such as NF-B, are limited or inactive, a condition presumably present in vivo. By interacting with Sp1, Vpr also has the potential to influence cellular gene expression and cellular functions. Thus, therapeutic approaches directed toward blocking the Vpr trans-activation function could prove valuable in treating AIDS. HIV-11 is the etiological agent of AIDS. The hallmark of AIDS is the slow but progressive depletion of CD4 ϩ -T cells, a class of T cells crucial for immune functions. Depletion of CD4 ϩ -T cell results in immunodeficiency and AIDS-related disorders, including encephalopathy, dementia, and malignancies (1). Despite tremendous efforts in the past, the mechanism of these AIDS-related disorders has remained unclear. However, it is clear that these are a consequence of function of HIV-1 encoded gene products. For example, the HIV-1 envelope glycoprotein was implicated to be involved in toxic effects on neuronal cells (2). Recently, the HIV-1 Vpr protein in peripheral blood of HIV-1-infected people was shown to activate HIV-1 replication in latently infected cells (3,4). This effect of Vpr was suggested to contribute to HIV-1 pathogenesis in vivo.The HIV-1 genome encodes structural as well as regulatory gene products (5, 6). Recently, great efforts have been made toward understanding the function of the so-called accessory regulatory genes, namely vif, vpr, vpu, and nef. These genes are generally non-essential for HIV-1 to replicate in activated T cells. Yet, animal model studies with two of these genes, vpr and nef, suggested that they are required for in vivo replication and pathogenesis of the simian immunodeficiency virus (7,8). The paradox between HIV-1 replication in vitro and that in vivo suggests that HIV-1 replication may be subjected to different modes of regulation in vivo compared to in vitro. For example, in vitro studies have shown that HIV-1 replication is highly dependent on cellular activation and availability of activated NF-B transcription factor (5, 6, 9). However, in vivo, the majority of the sus...

By animal-to-animal passage in macaques we derived a pathogenic chimeric simian-human immunodeficiency virus (SHIV) that caused CD4+ T cell loss and AIDS in pigtail macaques and used it to inoculate 20 rhesus and pigtail macaques by the intravaginal and intravenous routes. On the basis of the outcome of infection and patterns of CD4+ T cell loss and viral load, disease was classified into four patterns: acute, subacute, chronic, and nonprogressive infection. During the study period, 15 of the 20 animals developed fatal disease, including AIDS, encephalitis, pneumonia, and severe anemia. Opportunistic pathogens identified in these animals included Pneumocystis, cytomegalovirus, Cryptosporidium, Toxoplasma, and Candida. No single parameter by itself predicted outcome, although a combination of low CD4+ T cell counts in blood, high plasma virus levels, and presence of autoantibodies to red blood cells reliably predicted a fatal outcome. Five animals (25%) died within 3 months of inoculation and constituted the group with acute disease, whereas the nine animals (45%) with subacute disease died between 3 and 8 months postinoculation. This 70% mortality within 8 months is significantly shorter than in HIV-1-infected human beings, of whom 70% develop fatal disease a decade after infection. SHIV infection in macaques provides a useful model with which to evaluate antiviral strategies, combining all the advantages of the SIVmac system, yet using a virus bearing the envelope gene of HIV-1.

We recently reported that a chimeric simian/human immunodeficiency virus (SHIVKU-1) developed in our laboratory caused progressive depletion of CD4+ T lymphocytes and AIDS within 6 months of inoculation into pig-tailed macaques (M. nemestrina). None of the pig-tailed macaques showed productive SHIV infection in the central nervous system (CNS). In this report, we show that by further passage of the pathogenic virus in rhesus macaques [M. mulatta], we have derived a new strain of SHIV (SHIVKU-2) that has caused AIDS and productive CNS infection in 3 of 5 rhesus macaques infected with the virus. Productive replication of SHIV in the CNS was clearly shown by high infectivity titers and p27 protein levels in brain homogenates, and in 2 of the 3 rhesus macaques this was associated with disseminated, nodular, demyelinating lesions, including focal multinucleated giant cell reaction, largely confined to the white matter. These findings were reminiscent of HIV-1 associated neurological disease, and our immunohistochemical and in situ hybridization data indicated that the neuropathological lesions were associated with the presence of SHIV-specific viral antigens and nucleic acid respectively. However, the concomitant reactivation of opportunistic infections in these macaques suggested that such pathogens may have influenced the replication of SHIV in the CNS, or modified the neuropathological sequelae of SHIV infection in the rhesus species, but not in pig-tailed macaques. Our findings in the two species of macaques highlight the complexities of lentiviral neuropathogenesis, the precise mechanisms of which are still elusive.

A chimeric simian-human immunodeficiency virus (SHIV-4) containing the tat, rev, vpu, and env genes of HIV type 1 (HIV-1) in a genetic background of SIVmac239 was used to develop an animal model in which a primate lentivirus expressing the HIV-1 envelope glycoprotein caused acquired immune deficiency syndrome (AIDS) in macaques. An SHIV-infected pig-tailed macaque that died from AIDS at 24 weeks postinoculation experienced two waves of viremia: one extending from weeks 2-8 and the second extending from week 18 until death. Virus (SHIVKU-1) isolated during the first wave was neutralized by antibodies appearing at the end of the first viremic phase, but the virus (SHIVKU-1b) isolated during the second viremic phase was not neutralized by these antibodies. Inoculation of SHIVKU-1b into 4 pig-tailed macaques resulted in severe CD4(+) T cell loss by 2 weeks postinoculation, and all 4 macaques died from AIDS at 23-34 weeks postinoculation. Because this virus had a neutralization-resistant phenotype, we sequenced the env gene and compared these sequences with those of the env gene of SHIVKU-1 and parental SHIV-4. With reference to SHIV-4, SHIVKU-1b had 18 and 6 consensus amino acid substitutions in the gp120 and gp41 regions of Env, respectively. These compared with 10 and 3 amino acid substitutions in the gp120 and gp41 regions of SHIVKU-1. Our data suggested that SHIVKU-1 and SHIVKU-1b probably evolved from a common ancestor but that SHIVKU-1b did not evolve from SHIVKU-1. A chimeric virus, SHIVKU-1bMC17, constructed with the consensus env from the SHIVKU-1b on a background of SHIV-4, confirmed that amino acid substitutions in Env were responsible for the neutralization-resistant phenotype. These results are consistent with the hypothesis that neutralizing antibodies induced by SHIVKU-1 in pig-tailed macaque resulted in the selection of a neutralization-resistant virus that was responsible for the second wave of viremia.

By subcutaneous inoculation of SHIV(KU-2) in the hands of macaques, we developed a model of human immunodeficiency virus type-1 (HIV-1) occupational infection due to needle-stick injury and used the model to determine whether neutralizing serum to SHIV administered before or after virus inoculation could either prevent or abort infection, respectively. Six rhesus macaques were given 15 ml/kg pooled anti-SHIV plasma and challenged 24 hr later with approximately 300 animal infectious doses of SHIV(KU-2), subcutaneously. Three of the six macaques completely resisted infection with SHIV(KU-2). A fourth animal failed to yield infectious virus, but DNA extracted from its peripheral blood mononuclear cells (PBMC) and lymph nodes had viral sequences. Partial resistance was noted in the other two animals because virus recovery was delayed compared with the control animals. In contrast, six of six macaques given the same dose of anti-SHIV plasma 18 hr after exposure to virus became infected, as did two of two macaques given anti-SHIV plasma only 2 hr after exposure to virus. Our results suggest that neutralizing antibodies may have a prophylactic but not a therapeutic role in HIV-1 infections.