Simian Immunodeficiency Virus Model Research Articles

Nonhuman primate models of NeuroAIDS

Human Immunodeficiency virus (HIV), the virus that causes acquired immunodeficiency syndrome (AIDS), also manifests neurological complications. HIV-associated dementia (HAD) is the most severe form of HIV-induced neurocognitive disorders. HIV encephalitis (HIVE), the pathological correlate of HAD, is characterized by the formation of multinucleated giant cells and microglial nodules, astrocytosis, and neuronal damage and loss. Pathological evaluation of HAD disease progression in humans is not possible, with the only data collected being from individuals who have succumbed to the disorder, a snap shot of end-stage disease at best. Therefore, pertinent animal models have been developed to alleviate this gap of knowledge in the field of neurovirology and neuroinflammation. In general, the most widely used animal models are the simian immunodeficiency virus (SIV) and the chimeric simian/human immunodeficiency virus (SHIV) macaque model systems. Although both SIV and SHIV model systems are able to potentiate neuroinvasion and the concomitant neuropathology similar to that seen in the human syndromes, the innate differences between the two in disease pathogenesis and progression make for two separate, yet effective, systems for the study of HIV-associated neuropathology.

Development and characterization of a new single cycle vaccine vector in the simian immunodeficiency virus model system

Development and characterization of a new single cycle vaccine vector in the simian immunodeficiency virus model system

Role of the gastrointestinal tract in establishing infection in primates and humans

Here we present recent studies examining the gastrointestinal tract during primary HIV and simian immunodeficiency virus infections and emphasizing the onset of severe pathologic processes that are not adequately reflected in peripheral blood. We discuss these findings and hypotheses relating to the role of the gastrointestinal tract in HIV-1 pathogenesis. High levels of viral replication in the gastrointestinal mucosa during primary HIV and simian immunodeficiency virus infections lead to severe depletion of effector memory CD4 T cells coinciding with increased immune activation and mucosal damage. Viral reservoirs established at this stage appear to be persistent over the course of infection and during therapy. In the simian immunodeficiency virus model of AIDS, onset of the impaired intestinal epithelial barrier function and renewal was observed during primary viral infection. Dysfunction of the mucosal immune system and the epithelial barrier may contribute to viral persistence and impaired responses to microbial pathogens in infected individuals. A better understanding of the impact of HIV infection on the mucosal immune system may help in the development of newer preventive and therapeutic strategies directed against the virus.

Development and characterization of a new single cycle vaccine vector in the simian immunodeficiency virus model system

We have developed a new single cycle lentiviral vector, SIVsmH4i-SC27.1, as a potential SIV/HIV-1 vaccine candidate. This viral vector is capable of expressing all of the SIV gene products but is limited to one round of infection. The vector was created by mutating 27 codons dispersed among the viral vif, env, and nef genes to block protein function, attenuate viral replication/infectivity, and reduce the ability of the virus to manipulate the host immune system. To complement the env and nef replication defects, SC27.1 was pseudotyped with the VSV G glycoprotein to allow particle entry. The vif mutation was complemented by producing particles from an APOBEC3G-negative cell line, and the Vif protein defect was validated by showing that the single cycle virus lost most of its infectivity when particles were produced in presence of APOBEC3G. To deal with the problem of an antibody response to the VSV G protein in a vaccination strategy, two additional serotypes of the VSV G protein were used to create pseudotyped virus particles, and we observed no cross-neutralization activity for two of the pseudotyped particles with a potent neutralizing antiserum to one of the VSV G proteins. We detected moderate inhibition of infectivity in normal human and macaque sera, especially to the New Jersey serotype of VSV G, but as a heat sensitive activity, presumably complement mediated. These particles can be used in a prime-boost strategy to determine if a single cycle lentiviral vaccine vector capable of expressing all of the viral gene products holds promise in inducing immunity and protection to an SIVsm challenge.

Antibody-Mediated Neutralization and Simian Immunodeficiency Virus Models of HIV/AIDS

Over the last two decades, use of SIV for experimental infection of Asian macaques has provided important leads in the quest for an AIDS vaccine, served as the genesis of recombinant SIV/HIV viruses (SHIV), and perhaps most importantly, helped establish or confirm biological relevance for a variety of hypotheses related to the host immune response to infection and the corresponding viral strategies for evading that response. The env genes of HIV-1 and SIV encode proteins bearing a high degree of structural similarity and sharing an identical suite of essential functions. The Env complexes formed by these proteins are present on the surface of virus-producing cells and virions, where they are the primary targets of the host neutralizing antibody response. In this review, we briefly describe the similarities between HIV-1 infection and SIV experimental models, then focus specifically on the use of the SIV/macaque model as a tool for understanding the humoral immune response to infection and resistance to antibody-mediated neutralization in HIV infection and AIDS.

Perspectives on interleukin-7 therapy in HIV infection

HIV infection is characterized by depletion of CD4 T cells and altered immune function, leading to severe immune deficiency. Mechanisms leading to this T-cell depletion are not completely understood. Potent antiretroviral therapy restores T-cell counts and improves prognosis. Apart from antiviral therapy for the infection, immunotherapies such as interleukin-7 that influence T-cell homeostatic mechanisms are undergoing clinical evaluation. Because of its pleiotropic effects on developing and mature T cells, interleukin-7 may help to restore immune function during HIV infection. Recent studies explored the therapeutic use of interleukin-7 in simian immunodeficiency virus models and in HIV-infected patients. Interleukin-7 can help to restore CD4 T-cell number and function. Numerous recent findings highlight the importance of interleukin-7 pathway impairment in the pathogenesis of HIV infection. Notably, interleukin-7 levels increased with advancing CD4 T-cell lymphopenia, whereas interleukin-7 receptor expression is downregulated mainly on CD8 T cells. Therapeutic trials conducted in monkeys and in humans (phase I) have provided evidence on the role of interleukin-7 in thymopoiesis and in restoration of T-cell functions. Interleukin-7 appeared to be well tolerated and to have no deleterious effects on viral load. These results should be confirmed in larger phase I/II studies.

Sequence variation in the CC-chemokine ligand 2 promoter of pigtailed macaques is not associated with the incidence or severity of neuropathology in a simian immunodeficiency virus model of human immunodeficiency virus central nervous system disease

Increased expression of CC-chemokine ligand 2 (CCL2) in the cerebrospinal fluid (CSF) and brain is consistently observed in human immunodeficiency virus (HIV) and simian immunodeficiency virus (SIV) central nervous system (CNS) disease. The molecular basis for the correlation between increased expression of CCL2 and HIV neuropathogenesis has been linked to a polymorphism at -2578 in the promoter of human CCL2, which was reported to influence the rate of progression to acquired immunodeficiency syndrome (AIDS) and the predisposition of HIV-infected individuals to develop HIV-associated dementia. However, because the rate of neurological deterioration essentially parallels the progression of immunosuppression, it is inherently difficult to uncouple the influence of this polymorphism on increased progression to AIDS from increased propensity to develop CNS complications. To further investigate the correlation between CCL2 and HIV/SIV CNS disease, the authors sequenced the CCL2 promoter of 29 pigtailed macaques examined in their accelerated and consistent SIV model in which all infected macaques develop AIDS but only 69% developed moderate/severe CNS lesions. Sequence analysis identified 39 sites of nucleotide variation in the pigtailed macaque CCL2 promoter/enhancer regions, with the resulting consensus sequence aligning with 94.7% homology to the human CCL2 promoter. After genetic analyses, no variation was found to correlate with the incidence or severity of CNS lesions or with levels of CCL2 in plasma or CSF. These findings suggest that the determinants of neuropathogenesis in this SIV model are distinct from variation in these regions of the CCL2 promoter.

Bioengineering Lactic Acid Bacteria to Secrete the HIV-1 Virucide Cyanovirin

An urgent need exists to prevent the sexual transmission of HIV-1. With prevalence rates exceeding 35% in parts of sub-Saharan Africa, increasing attention has been placed on developing and testing microbicidal agents capable of preventing virus transmission at mucosal sites. HIV-1 microbicides must meet several requirements before their widespread use. The drugs must be able to neutralize a diversity of HIV-1 strains, not induce mucosal inflammation, be associated with minimal side effects, and be effective for a prolonged period after a single application. Recent work has demonstrated the utility of recombinant lactic acid bacteria (LAB) as agents of mucosal drug delivery. Here, we describe the bioengineering of strains of LAB to secrete the prototypic virucidal compound cyanovirin (CV-N) and demonstrate the anti-HIV-1 activity of secreted CV-N. Our results suggest that recombinant LAB may serve as effective microbicidal compounds and deserve in vivo testing in simian immunodeficiency virus models of mucosal virus transmission.

Inhibition of simian immunodeficiency virus by foamy virus vectors expressing siRNAs

Viral vectors available for gene therapy are either inefficient or suffer from safety concerns for human applications. Foamy viruses are non-pathogenic retroviruses that offer several unique opportunities for gene transfer in various cell types from different species. In this report, we describe the use of simian foamy virus type 1 (SFV-1) vector to examine the efficacy of therapeutic genes. Hairpin short-interfering RNA (siRNA) that targets the simian immunodeficiency virus (SIV) rev/env was placed under the control of the PolIII U6 snRNA promoter for expression and screened for silencing target genes using cognate target–reporter fusions. We have identified an effective siRNA (designated R2) which reduces the rev and env gene expression by 89% and 95%, respectively. Using the simian foamy virus type 1 (SFV-1) based vector, we delivered the PolIII expressed R2 siRNA into cultured cells and challenged with SIV. The results show that the R2 siRNA is a potent inhibitor of SIV replication as determined by p27 expression and reverse transcriptase assays. Vectors based on a non-pathogenic SFV-1 vector may provide a safe and efficient alternative to currently available vectors, and the SIV model will help devise protocols for effective anti-HIV gene therapy.

Evolution of nef variants in gut associated lymphoid tissue of rhesus macaques during primary simian immunodeficiency virus infection

We utilized the simian immunodeficiency virus model of AIDS to examine evolution of nef gene in gut-associated lymphoid tissue (GALT) during primary and early asymptomatic stages of infection. Macaques were infected with a cloned virus, SIVmac239/nef-stop harboring a premature stop codon in the nef gene. Restoration of the nef open reading frame occurred in GALT early at 3 days post-infection. Analysis of nef sequences by phylogenetic tools showed that evolution of nef was neutral thereafter, as evidenced by the ratio of synonymous to nonsynonymous substitutions, a star pattern in unrooted trees and distribution of amino acid replacements fitting a simple Poisson process. Two regions encoding for a nuclear localization signal and a CTL epitope were conserved. Thus, GALT was a site for strong positive selection of functional nef during initial stages of infection. However, evolution of the nef gene thereafter was neutral during early asymptomatic stage of infection.

477. Development of SIV Based Vectors with Efficient Delivery for Vector Dynamic Studies in the Simian System

Lentiviruses are well known for their species specificity, and consequently lentivirus-derived vectors exhibit the highest gene transfer efficiency in cells of the same species from which the vectors were derived. Therefore, to accurately model a gene therapy approach for HIV in a monkey model, a parallel simian immunodeficiency virus (SIV) vector should be developed to accurately model the vector-cell-virus interactions. Our first therapeutic HIV-derived vector VRX496 contains an antisense targeting HIV env region. We established the safety of this novel vector in a Phase I open label clinical trial, which was allowed to proceed without preliminary studies in monkeys due to 1) the engineered safety features of the vector and 2) the observation that a monkey study would not be informative regarding the dynamics of an HIV-based vector. After demonstrating safety of this vector with early indications of efficacy in humans, we hypothesized that subsequent generations of this vector that have an improved ability to mobilize may increase the therapeutic efficacy of our anti-HIV gene therapy. However, such mobilizing vectors are considered too risky to test directly in patients, due to concerns regarding insertional oncogenesis. Therefore, we chose to test this hypothesis in monkeys in order to safely model the dynamics of these new vectors in vivo. Accordingly, we developed three SIVmac239 based lentiviral vectors, one that parallels our clinical vector VRX496 that contains no viral proteins (pN1), a second generation (pN2) containing the gag-pol protein that may have an improved ability to mobilize as a result of reduced dependency on wtSIV-proteins, and a third (empty pN2) containing gag-pol and no antisense payload to serve as a control. We demonstrated that the SIV based vectors were able to transduce the primary simian CD4 cells significantly better than the counterpart of HIV based vector by an approximately 6-fold improvement in gene transfer. We also showed that the vector potently inhibited the SIVmac239 replication in both transduced CEMx174 cells and primary simian CD4 cells to below the limit of detection, as previously observed with VRX496 and HIV in primary human CD4 cells. Detailed analysis of these results and modelling of these vectors in a monkey SIV model will be discussed.

Neuroprotective and Anti–Human Immunodeficiency Virus Activity of Minocycline

The prevalence of human immunodeficiency virus (HIV) central nervous system (CNS) disease has not decreased despite highly active antiretroviral therapy. Current antiretroviral drugs are expensive, have significant adverse effects including neurotoxicity, and few cross the blood-brain barrier. To examine the ability of minocycline, an antibiotic with potent anti-inflammatory and neuroprotective properties, to protect against encephalitis and neurodegeneration using a rapid, high viral load simian immunodeficiency virus (SIV) model of HIV-associated CNS disease that constitutes a rigorous in vivo test for potential therapeutics. Five SIV-infected pigtailed macaques were treated with 4 mg/kg per day of minocycline beginning at early asymptomatic infection (21 days after inoculation). Another 6 macaques were inoculated with SIV but remained untreated. Blood and cerebrospinal fluid (CSF) samples were taken on days 7, 10, 14, 21, 28, 35, 43, 56, 70, 77, and 84, and all macaques were humanely killed at 84 days after inoculation, a time that corresponds to late-stage infection in HIV-infected individuals. Blood and CSF samples were tested for viral load by real-time reverse transcription-polymerase chain reaction and levels of monocyte chemoattractant protein 1 were quantitated by enzyme-linked immunosorbent assay. The presence and severity of encephalitis was determined by microscopic examination of tissues. Central nervous system inflammation was further assessed by measuring infiltration and activation of macrophages, activation of p38 mitogen-activated protein kinase and expression of amyloid precursor protein by quantitative immunohistochemistry. Minocycline-treated macaques had less severe encephalitis (P = .02), reduced CNS expression of neuroinflammatory markers (major histocompatibility complex class II, P = .03; macrophage marker CD68 , P = .07; T-cell intracytoplasmic antigen 1, P = .03; CSF monocyte chemoattractant protein 1, P = .001), reduced activation of p38 mitogen-activated protein kinase (P<.001), less axonal degeneration (beta-amyloid precursor protein, P = .03), and lower CNS virus replication (viral RNA, P = .04; viral antigen, P = .04). In in vitro analysis, minocycline suppression of HIV and SIV replication in cultured primary macrophages did not correlate with suppression of activation of p38-mitogen-activated protein kinase pathways, whereas suppression in primary lymphocytes correlated with suppression of p38 activation. In this experimental SIV model of HIV CNS disease, minocycline reduced the severity of encephalitis, suppressed viral load in the brain, and decreased the expression of CNS inflammatory markers. In vitro, minocycline inhibited SIV and HIV replication. These findings suggest that minocycline, a safe, inexpensive, and readily available antibiotic should be investigated as an anti-HIV therapeutic.

Novel Adeno-Associated Virus Vector Vaccine Restricts Replication of Simian Immunodeficiency Virus in Macaques

Gene transfer vectors based on recombinant adeno-associated virus (rAAV) are simple, versatile, and safe. While the conventional applications for rAAV vectors have focused on delivery of therapeutic genes, we have developed the system for delivery of vaccine antigens. In particular, we are interested in generating rAAV vectors for use as a prophylactic human immunodeficiency virus type 1 (HIV-1) vaccine. To that end, we constructed vaccine vectors that expressed genes from the simian immunodeficiency virus (SIV) for evaluation in the monkey SIV model. After a single intramuscular dose, rAAV/SIV vaccines elicited SIV-specific T cells and antibodies in macaques. Furthermore, immunized animals were able to significantly restrict replication of a live, virulent SIV challenge. These data suggest that rAAV vaccine vectors induced biologically relevant immune responses, and thus, warrant continued development as a viable HIV-1 vaccine candidate.

Severe subcortical degeneration in macaques infected with neurovirulent simian immunodeficiency virus

Infection with human immunodeficiency virus-1 (HIV-1), the causative agent of acquired immunodeficiency syndrome (AIDS) in humans, causes a spectrum of neuropathology that includes alterations in behavior, changes in evoked potentials, and neuronal degeneration. In the simian immunodeficiency virus (SIV) model of HIV infection, affected monkeys show clinical symptoms and neurological complications that mimic those observed in human neuro-AIDS. To investigate the relationship between morphological correlates and neurophysiological deficits, unbiased stereology was used to assess total neuron number, volume, and neuronal density for all neurons in the globus pallidus (GP) and for dopamine (DA)-containing neurons in the substantia nigra (SN) in eight macaques inoculated with macrophage-tropic, neurovirulent SIV (SIVmac R71/17E), and five control animals. There was a significant difference between rapid progressors and controls for both neuron number (P < .01) and neuronal density (P < .05) in the GP, and for neuron number (P < .05) in the SN. Neuron loss ranged from 6% to 70% in the GP and from 10% to 50% in the SN. Neuropathological analyses confirmed neuroAIDS-like changes in brain, including microglial nodules, extensive perivascular cuffing and/or the presence of multinucleated giant cells, and alterations in neuronal morphology in the majority of the rapid progressors. By comparison, slow progressors showed little, if any, neuropathology. These neuropathological changes in SIV-infected monkeys indicate that neuron death and morphological alterations in the basal ganglia may contribute to the motor impairments reported in the SIV model and, by analogy, in the subset of patients afflicted with motor impairment in human neuro-AIDS.

Reversion of the M184V mutation in simian immunodeficiency virus reverse transcriptase is selected by tenofovir, even in the presence of lamivudine.

The methionine-to-valine mutation in codon 184 (M184V) in reverse transcriptase (RT) of human immunodeficiency virus type 1 (HIV-1) or simian immunodeficiency virus (SIV) confers resistance to (-)-2'-deoxy-3'-thiacytidine (3TC; lamivudine) and increased sensitivity to 9-[2-(phosphonomethoxy)propyl]adenine (PMPA; tenofovir). We have used the SIV model to evaluate the effect of the M184V mutation on the emergence of resistance to the combination of 3TC plus PMPA. A site-directed mutant of SIVmac239 containing M184V (SIVmac239-184V) was used to select for resistance to both 3TC and PMPA by serial passage in the presence of increasing concentrations of both drugs. Under these selection conditions, the M184V mutation reverted in the majority of the selections. Variants resistant to both drugs were found to have the lysine-to-arginine mutation at codon 65 (K65R), which has previously been associated with resistance to PMPA in both SIV and HIV. Similarly, in rhesus macaques infected with SIVmac239-184V for 46 weeks and treated daily with (-)-2'-deoxy-5-fluoro-3'-thiacytidine [(-)-FTC], there was no reversion of M184V, but this mutation reverted to 184 M in all three animals within 24 weeks of treatment with (-)-FTC and PMPA. Although the addition of PMPA to the (-)-FTC therapy induced a decrease in virus loads in plasma, these loads eventually returned to pre-PMPA levels in each case. All animals receiving this combination developed the K65R mutation. These results demonstrate that the combination of PMPA with 3TC or (-)-FTC selects for the K65R mutation and against the M184V mutation in SIV RT.

Central Nervous System Correlates of Behavioral Deficits Following Simian Immunodeficiency Virus Infection

Despite the high incidence of cognitive and motor impairment in acquired immunodeficiency syndrome (AIDS) patients, the mechanisms of AIDS-related central nervous system (CNS) pathology are not completely understood. Infection with simian immunodeficiency virus (SIV) in macaques provides an excellent model of AIDS, including human immunodeficiency virus (HIV)-induced CNS pathology and cognitive/behavioral impairment. Co-inoculation with two SIV strains, SIV/17E-Fr and SIV/DeltaB670, accelerates SIV CNS disease, producing SIV encephalitis in over 90% of pig-tailed macaques within 3 months. In the present study, this SIV model was employed to identify cellular and viral correlates of behavioral impairment following SIV infection. Measures of psychomotor speed (simple reaction time), fine motor control (bimanual motor task), and general motor activity (home cage movement) were all adversely affected by SIV disease. Prior to euthanasia, performance was significantly impaired in both a simple reaction time task in 6 of 12 monkeys and a bimanual motor task in 5 of 6 monkeys. All monkeys evaluated (11 of 11) showed significant reductions in spontaneous motor activity. Significant correlations were found between impaired performance on the bimanual motor test and axonal damage (accumulation of beta-amyloid precursor protein in the corpus callosum) as well as increased microglial activation and macrophage infiltration (levels of CD68 and Ham56 immunostaining). These results suggest that axonal damage is related to the behavioral impairment induced by infection with SIV. The axonal damage may result from neuroimmune responses, including microglial and macrophage activation. Therefore, axonal damage may be a morphologic manifestation of neuronal dysfunction that underlies development of behavioral impairment in HIV/SIV CNS infection.

A novel simian immunodeficiency virus model that provides insight into mechanisms of human immunodeficiency virus central nervous system disease.

Although highly active antiretroviral therapy (HAART) has reduced the incidence of dementia, a significant proportion of HIV-infected individuals cease therapy because of unacceptable side effects. Further, HAART is not available to the majority of HIV-infected people worldwide. Thus, animal models remain an important means by which to investigate the pathogenesis of HIV-induced CNS disease, particularly events during acute infection. We have developed an accelerated, consistent SIV/macaque model of HIV infection in which over 90 percent of inoculated macaques develop SIV encephalitis and behavioral changes by three months post-inoculation (p.i.). Viral load in the CSF during terminal infection and the ratio of MCP-1 in CSF:plasma are strong predictors of the severity of encephalitis at necropsy. The high incidence of CNS disease in this model provides an opportunity to correlate host and viral events in the CNS during acute infection with the later development of CNS disease. Using this model, we have demonstrated that viral DNA persists in the brain throughout asymptomatic infection despite significant suppression of active virus replication. We have further demonstrated that encephalitis is associated with the selective replication of macrophage-tropic, neurovirulent viral strains, and that recrudescence of virus replication during terminal infection occurs by reactivation of preexisting viral strains in the brain.

Simian immunodeficiency virus model of HIV induced central nervous system dysfunction

Publisher Summary This chapter discusses the advances that have been made in the development and refinement of the simian immunodeficiency virus (SIV) model of human immunodeficiency virus (HIV) dementia, and its contribution to the understanding and management of the acquired immunodeficiency syndrome (AIDS) dementia complex (ADC) in humans. The SIV and the virus HIV are primate lentiviruses that have great genomic, structural, and virologic similarities. SIVs have been isolated from the different genera of nonhuman primates and hence are designated by subscripts indicating the type of primate from which they originally were isolated—for example, SIV mac from rhesus monkeys and SIV smm from sooty mangabey monkeys. Infection of rhesus as well as pig-tailed monkeys is the most popular nonhuman primate model of HIV disease. Progressive lymphoid changes, opportunistic infections, a wasting syndrome, and central nervous system (CNS) disease characterize the syndrome in both humans and monkeys. In humans, the HIV-associated cognitive/motor disorder, also known as ADC or neuroAIDS occurs in one-third of the patients. The etiology of CNS dysfunction resulting from HIV and SIV infections likely depends on host as well as viral factors. The SIV/nonhuman primate model of the HIV-associated cognitive/motor disorders provides a splendid system for studying this interplay.

Paradigms for behavioral assessment of of viral pathogenesis

Publisher Summary This chapter uses two approaches to describe the behavioral paradigms used in assessing viral pathology. First approach is the behavioral effects of viral infection in rodents by providing a broad overview of the types of studies conducted in rodents and by using examples from a number of virus model systems. Secondly, it discusses the behavioral effects of viral infection in nonhuman primates by focusing on the simian immunodeficiency virus model of acquired immunodeficiency syndrome and elaborating on recent work involving a number of behavioral paradigms in this one model system. Neurobehavioral dysfunction is a serious consequence of a number of viral infections in which the pathological and toxic changes induced by the virus affect nervous system structure and function. Viral infections have been associated with a wide range of functional and behavioral abnormalities, including those that affect basic physiology and homeostasis, motor performance, affective behavior, and complex cognition. Various neurotropic viruses may trigger transient or irreversible disorders in the central nervous system by causing neuronal death, altering the expression of neurotransmitters, neuropeptides, or receptors or by triggering immune system mechanisms that interfere with neural function. One primary objective of the neurobehavioral studies of viral pathogenesis is to characterize specific behavioral phenotypes in infection models, in both rodents and nonhuman primates, that can then be correlated with the distinctive neuropathology and immunopathology observed in these same models.

New prospects for the development of a vaccine against human immunodeficiency virus type 1. An overview

During the past few years, definite progress has been made in the field of human immunodeficiency virus type 1 (HIV-1) vaccines. Initial attempts using envelope gp120 or gp1 40 from T-cell line-adapted (TCLA) HIV-1 strains to vaccinate chimpanzees showed that neutralizing antibody-based immune responses were protective against challenge with homologous TCLA virus strains or strains with low replicative capacity, but these neutralizing antibodies remained inactive when tested on primary HIV-1 isolates, casting doubts on the efficacy of gp120-based vaccines in the natural setting. Development of a live attenuated simian immunodeficiency virus (SIV) vaccine was undertaken in the macaque model using whole live SIV bearing multiple deletions in the nef, vpr and vpx genes. This vaccine provided remarkable protective efficacy against wild-type SIV challenge, but the deletion mutants remain pathogenic, notably in neonate monkeys. Study of the mechanisms of protection in the SIV model unravelled the importance of the T-cell responses, whether in the form of cytotoxic T-lymphocyte (CTL) killing activity, or in that of antiviral factor secretion of cytokines, β-chemokines and other unidentified antiviral factors by CD8 + T-cells. Induction of such a response is being sought at this time using various live recombinant virus vaccines, either poxvirus or alphavirus vectors or DNA vectors, which can be combined together or with a gp120/ gp140 boost in various prime-boost combination strategies. New vectors include attenuated vaccinia virus NYVAC, modified vaccinia strain Ankara (MVA), Semliki Forest virus, Venezuelan equine encephalitis virus, and Salmonellas. Recent DNA prime-poxvirus boost combination regimens have generated promising protection results against SIV or SIV/HIV (SHIV) challenge in macaque models. Emphasis is also put on the induction of a mucosal immune response, involving both a secretory IgA response and a mucosal CTL response which could constitute a ‘first line of defence’ in the vaccinated host. Finally, a totally novel vaccine approach based on the use of Tat or Tat and Rev antigens has been shown to induce efficient protection from challenge with pathogenic SIV or SHIV in vaccinated macaques. The only vaccine in phase 3 clinical trials in human volunteers is a gp120-based vaccine, AIDSVAX TM. A prime-boost combination of a recombinant canarypoxvirus and a subunit gp120 vaccine is in phase 2. Emphasis has been put recently on the necessity of testing prototype vaccines in developing countries using immunogens derived from local virus strains. Trial sites have thus been identified in Kenya, Uganda, Thailand and South Africa where phase I trials have begun or are expected to start presently.