HIV-2 Envelope Protein Research Articles

English

Serological methods are the main methods in diagnosis of human immunodeficiency virus (HIV)-1 and HIV-2 infection. The western Blot (WB) is considered as a gold standard test for the confirmation of enzyme-linked immunosorbent assay (ELISA) and/or other rapid screened reactive results samples in the diagnosis of HIV infection. However, problems of WB, particularly in regard to the significance of indeterminate results, still remain. To develop and evaluate a recombinant HIV-1/2 antigens-based line immunoassay as confirmatory assay for HIV-1 and HIV-2 infections is the objective of this study. HIV-1 envelope proteins (gp160, gp120, gp41), HIV-1 integrase protein (p31), HIV-1 gag proteins (p24, p17) and HIV-2 envelope protein (gp 36) were cloned and expressed inEscherichia coli. All recombinant HIV antigens were purified and immobilized on nitrocellulose membranes and applied in a line immunoassay, which supplement detected anti-HIV antibodies in a single strip. Specific antibodies to these antigens in human sera were revealed using an alkaline phosphatase labeled anti-human immunoglobulin G (IgG) as second antibody. Two anti-human IgG lines were used as a control for the addition of serum. Positive or negative results were determined by visual comparison of the antigen lines intensity with the two control lines. The recombinant HIV antigens-based line immunoassay identified seropositive individuals with a high degree of accuracy; none of the HIV-seropositive subjects yielded a negative result. Reactivity with these antigens, demonstrated 100% sensitivity and specificity in distinguishing seronegative from seropositive sera. This line immunoassay was much more sensitive and specific than WB. In particular, the line immunoassay allowed for a significant reduction in the number of indeterminate results and for more accurate distinction between HIV-1 and/or HIV-2 infections.   Key words: HIV antibodies, line immunoassay, recombinant antigens, confirmatory test.

Dendritic Cell Differentiation and Maturation in the Presence of HIV Type 2 Envelope

Dendritic cells (DCs) are fundamental for the initiation of immune responses and are important players in AIDS immunopathogenesis. Impairment of DC function may result from bystander effects of HIV-1 envelope proteins independently of direct HIV-1 infection. HIV-2 envelope proteins are thought to interact with a broader range of receptors than those of HIV-1, and have been shown to have T cell immunosuppressive properties mediated by monocytes. The effects of HIV-2 envelope on DC differentiation and maturation were investigated. The modulatory properties of the HIV-2 envelope on DC generated from monocytes were assessed using both recombinant proteins (HIV-2(ROD) and HIV-2(ALI)) and whole chemically inactivated virus (aldrithiol-2-treated HIV-2(ROD)). DC phenotype was assessed by flow cytometry and DC function by their ability to stimulate allogeneic T cells and to produce cytokines. We demonstrate that HIV-2 Env had no effects upon DC differentiation and maturation despite its broad receptor usage and ability to modulate monocyte function. It is plausible to speculate that a reduced ability of the HIV-2 Env to impair myeloid DC function could represent a contributory factor to the relatively benign course of HIV-2 disease.

Monocyte‐mediated T cell suppression by HIV‐2 envelope proteins

HIV-2 is associated with an attenuated form of HIV disease. We investigate here the immunosuppressive effects of the HIV-2 envelope protein, gp105. We found that gp105 suppresses activation of T cells through a monocyte-mediated mechanism. Suppression of T cell activation by gp105 depends on contact between monocytes and T cells, but not on CD4+CD25+ T cells. The TLR4 pathway is likely involved, since gp105 activates TLR4 signaling and induces TNF-alpha production by monocytes. Immunosuppression is viewed as the main pathophysiologic consequence of infection by HIV. However, the main immunologic defect caused by HIV, depletion of T cells, requires T cell activation. Our findings are consistent with a new concept that HIV-2 envelope proteins act on monocytes to suppress T cell activation and that this property may contribute to the benign course of HIV-2. We hypothesize that the HIV-2 envelope immunosuppressive properties limit bursts of T cell activation, thus reducing viremia and contributing to the slow rate of disease progression that characterizes HIV-2 disease.

Human TRIM5α mediated restriction of different HIV-1 subtypes and Lv2 sensitive and insensitive HIV-2 variants

In order to characterize the antiviral activity of human TRIM5α in more detail human derived indicator cell lines over expressing wild type human TRIM5α were generated and challenged with HIV-1 and HIV-2 viruses pseudotyped with HIV envelope proteins in comparison to VSV-G pseudotyped particles. HIV envelope protein pseudotyped particles (HIV-1[NL4.3], HIV-1[BaL]) showed a similar restriction to infection (12 fold inhibition) compared to VSV-G pseudotyped viruses after challenging TZM-huTRIM5α cells. For HIV-2 a stronger restriction to infection was observed when the homologous envelope protein Env42S was pseudotyped onto these particles compared to VSV-G pseudotyped HIV-2 particles (8.6 fold inhibition versus 3.4 fold inhibition). It has been shown that HIV-2 is restricted by the restriction factor Lv2, acting on capsid like TRIM5α. A mutation of amino acid 73 (I73V) of HIV-2 capsid renders this virus Lv2-insensitive. Lv2-insensitive VSV-G pseudotyped HIV-2/I73V particles showed a similar restriction to infection as did HIV-2[VSV-G] particles (4 fold inhibition). HIV-2 envelope protein (Env42S)-pseudotyped HIV-2/I73V particles revealed a 9.3 fold increase in infection in TZM cells but remained restricted in TZM-huTRIM5α cells (80.6 fold inhibition) clearly indicating that at least two restriction factors, TRIM5α and Lv2, act on incoming HIV-2 particles. Further challenge experiments using primary isolates from different HIV-1 subtypes and from HIV-1 group O showed that wild type human TRIM5α restricted infection independent of coreceptor use of the infecting particle but to variable degrees (between 1.2 and 19.6 fold restriction).

Marked immunosuppressive effects of the HIV-2 envelope protein in spite of the lower HIV-2 pathogenicity.

HIV-1 envelope proteins have immunosuppressive properties and it is thought that they have a role in the establishment of immunodeficiency. This study characterizes the immunological effects of HIV-2 envelope protein gp105, a virus which is associated with a slower rate of disease progression. The effects of recombinant baculovirus-expressed envelope proteins from HIV-IIIB HIV-1MN, HIV-2ROD and SIVmac251 on anti-CD3-stimulated peripheral blood mononuclear cells (PBMC) from healthy donors were evaluated by incorporation of 3H-thymidine, flow cytometric analysis of bromodeoxyuridine incorporation in different T cell subsets, kinetics of expression of costimulatory molecules (CD40L/OX40) and assessment of cell death by annexin V/propidium iodide staining. The effects on production of tumour necrosis factor alpha (TNF-alpha) by monocytes were assessed at the single-cell level after a 6 h culture of unstimulated PBMC. HIV-2 gp105 was more inhibitory than HIV-1 gp120 of T cell proliferation and the upregulation of CD40L and OX40; in the absence of signficant induction of apoptosis. This inhibition affected both CD4 and CD8 T cells and was only partially reversed by costimulation with interleukin 2 or CD28. gp105 strongly inducted TNF-alpha production by monocytes. The immunosuppressive properties of the HIV envelope proteins could be beneficial rather than detrimental to the host by interfering with the heightened state of immunocellular activation that characterizes HIV infection and by limiting the bursts of viral replication. This hypothesis could in part explain the slower decline of CD4 cell numbers in HIV-2 infection and deserves further exploration.

The use of a chimera HIV-1/HIV-2 envelope protein for immunodiagnosis of HIV infection: its expression and purification in E. coli by use of a translation initiation site within HIV-1 env gene.

A chimera HIV-1/HIV-2 envelope sequence composed of multiple conserved immunodominant epitopes of HIV-1 envelope protein (HIV-1 IIIB: env482-518 + env548-675) and the HIV-2 gp36 immunodominant epitope (env592-603), was constructed and directly over-expressed in E. coli by using a prokaryotic translation initiation sequence contained within the gene of HIV-1 envelope. The recombinant product was purified and applied in antibody-screening assay. The purified chimera antigen reacted with all the thirty-eight HIV-1 positive serum samples, the two HIV-2 serum samples, and had no cross-reaction with all the eighty-eight normal healthy serum sample. The results indicated that this recombinant chimera HIV-1/HIV-2 envelope protein could be useful for diagnostic purposes of HIV infection.

Simian immunodeficiency virus infection in a patas monkey (Erythrocebus patas): evidence for cross-species transmission from African green monkeys (Cercopithecus aethiops sabaeus) in the wild.

Socio-ethological studies on troops of African green monkeys (AGMs) (Cercopithecus aethiops sabaeus) and patas monkeys (Erythrocebus patas) in Senegal have documented physical contacts between these two species. Elevated simian immunodeficiency virus (SIV) seroprevalence rates have been reported for the different AGM subspecies. We report here the extent to which patas monkeys are infected and compare the relatedness of the viruses isolated from theses two different species. Among the 85 AGMs and 54 patas monkeys studied, 47% of 7.5%, respectively, had antibodies that cross-reacted with HIV-2 envelope proteins. From two AGMs a virus was isolated. From the patas monkeys, virus isolation was generally not possible, but from one animal that was ill a virus designated pamG31 was amplified by PCR. In addition, for the two SIVagm isolates, an 830 bp region spanning the env and nef genes was amplified and sequenced. Comparisons of sequences from the env/nef region revealed 80% identity between pam G31 and SIVagm isolates from AGMs of the sabaeus subspecies, and 94% identity between the two SIVagm isolates. Phylogenetic analysis showed that pamG31 belongs to the SIVagm sabaeus subgroup. This is the first report of a lentiviral infection in a patas monkey. The close genetic relatedness between pamG31 and SIVagm sabaeus viruses is a strong argument in favour of cross-species transmission of SIV between AGMs and patas monkeys in the wild. For these reasons, we propose to refer to this patas virus as SIVagm-pamG31.

Characterization of a Nuclear Retention Sequence within the 3′ Region of the HIV-2 Envelope Gene

During the phenotypic analysis of mutations in the HIV-2 envelope protein (gp140), we have identified a mutant that expresses high levels of gp140 in the absence of the Rev protein. This mutant was obtained by deletion of a 190-base pair fragment at the end of the env coding region. Northern blot analysis of RNAs produced by transfected HeLa cells showed that this fragment inhibits the cytoplasmic transport of unspliced env-specific RNAs in the absence of Rev. The "inhibitory element" was only active in the sense orientation and appeared to be specific for unspliced HIV-2 env RNAs, suggesting that it may have an important regulatory role for HIV-2 expression.

New peptides with immunological properties of HIV-2 envelope protein with the structure of simian immune deficiency virus proteins, useful in dignosis and as vaccine components

New peptides with immunological properties of HIV-2 envelope protein with the structure of simian immune deficiency virus proteins, useful in dignosis and as vaccine components

Distinction between HIV-1 and HIV-2 infection using novel synthetic lipopeptide conjugates as antigens in enzyme immunoassays

A novel immunoassay technique using synthetic lipopeptide (Pam 3Cys-Ser) linked to immunodominant peptide domains of HIV-1 and HIV-2 envelope proteins as an antigen adsorbent has been developed. Attachment of peptides to microtiter plates can be considerably improved with this method by employing the hydrophobic properties of lipopeptide. From the sera of 121 HIV-1 infected patients 117 reacted with Pam 3Cys-Ser-[HIV-1(598–609)cyclic disulfide]. Five of 5 HIV-2 positive sera were positive with Pam 3Cys-Ser-[HIV-2(593–603)cyclic disulfide]. Control sera failed to react with these conjugates.