Primary Human Leukocyte Subsets Differentially Express Vaccinia Virus Receptors Enriched in Lipid Rafts

Byrd, Daniel; Amet, Tohti; Hu, Ningjie; Lan, Jie; Hu, Sishun; Yu, Qigui

doi:10.1128/jvi.01545-13

Cited by 16 publications

(26 citation statements)

References 45 publications

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“…Cells were fixed with 2% paraformaldehyde (PFA), and then subjected to immunostaining for confocal microscopy analysis of viral protein expression and colocalization of HIV-1 Env with CD59 and ganglioside M1 (GM1) lipid rafts on the cell surface. Cells were incubated with cholera toxin subunit B (CTB) conjugated with Alexa Fluor 488 (Life Technologies, Carlsbad, CA) at 4°C for 20 min to stain GM1 lipid rafts as previously described (50). Anti-HIV-1 gp120 mAb cloned from an HIV-1-positive individual (2G12, NIH AIDS Reagent Program, Germantown, MD) and BRIC229 were used as primary Abs to stain Env and CD59, respectively.…”

Section: Methodsmentioning

confidence: 99%

Provirus Activation Plus CD59 Blockage Triggers Antibody-Dependent Complement-Mediated Lysis of Latently HIV-1–Infected Cells

Lan

Yang

Byrd

et al. 2014

The Journal of Immunology

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Latently HIV-1-infected cells are recognized as the last barrier towards viral eradication and cure. To purge these cells, we combined a provirus stimulant with a blocker of human CD59, a key member of the regulators of complement activation (RCA), to trigger antibody-dependent complement-mediated lysis (ADCML). Provirus stimulants including prostratin and histone deacetylase inhibitors (HDACi) such as romidepsin (RMD) and suberoylanilide hydroxamic acid (SAHA) activated proviruses in the latently HIV-1-infected T cell line ACH-2 as virion production and viral protein expression on the cell surface were induced. RMD was the most attractive provirus stimulant as it effectively activated proviruses at nanomolar concentrations that can be achieved clinically. Antiretroviral drugs including two protease inhibitors (atazanavir and darunavir) and a reverse transcriptase inhibitor (emtricitabine) did not affect the activity of provirus stimulants in the activation of proviruses. However, saquinavir (a protease inhibitor) markedly suppressed virus production, although did not affect the percentage of cells expressing viral Env on the cell surface. Provirus-activated ACH-2 cells expressed HIV-1 Env that colocalized with CD59 in lipid rafts on the cell surface, facilitating direct interaction between them. Blockage of CD59 rendered provirus-activated ACH-2 cells and primary human CD4+ T cells that were latently infected with HIV-1 sensitive to ADCML by anti-HIV-1 polyclonal antibodies or plasma from HIV-1-infected patients. Therefore, a combination of provirus stimulants with RCA blockers represents a novel approach to eliminate HIV-1.

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Section: Methodsmentioning

confidence: 99%

Provirus Activation Plus CD59 Blockage Triggers Antibody-Dependent Complement-Mediated Lysis of Latently HIV-1–Infected Cells

Lan

Yang

Byrd

et al. 2014

The Journal of Immunology

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“…ACH-2 cells were treated with PMA (10 ng/ ml)/ionomycin (1 g/ml) or DMSO for 24 h. Cells were fixed with 2% paraformaldehyde and then subjected to immunostaining for confocal microscopy analysis of viral protein expression and colocalization of HIV-1 Env with hCD59 and ganglioside M1 (GM1) lipid rafts on the cell surface. Cells were incubated with cholera toxin subunit B (CTB) conjugated with Alexa Fluor 488 (Life Technologies, Carlsbad, CA) at 4°C for 20 min to stain GM1 lipid rafts as previously described (49). Cells were also subjected to staining with an individual anti-HIV-1 Env MAb (1 g/ml) of nAbs (2G12, 2F5, and 4E10), non-nAbs (2.2C, A32, N5-i5, and N12-i15), or anti-hCD59 BRIC229.…”

Section: Methodsmentioning

confidence: 99%

Blockage of CD59 Function Restores Activities of Neutralizing and Nonneutralizing Antibodies in Triggering Antibody-Dependent Complement-Mediated Lysis of HIV-1 Virions and Provirus-Activated Latently Infected Cells

Yang

Lan

Shepherd

et al. 2015

J Virol

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Both HIV-1 virions and infected cells use their surface regulators of complement activation (RCA) to resist antibody-dependent complement-mediated lysis (ADCML). Blockage of the biological function of RCA members, particularly CD59 (a key RCA member that controls formation of the membrane attack complex at the terminal stage of the complement activation cascades via all three activation pathways), has rendered both HIV-1 virions and infected cells sensitive to ADCML mediated by anti-Env antibodies (Abs) or sera/plasma from patients at different stages of viral infection. In the current study, we used the well-characterized anti-HIV-1 neutralizing Abs (nAbs), including 2G12, 2F5, and 4E10, and non-nAbs, including 2.2C, A32, N5-i5, and N12-i15, to investigate whether the enhancement of ADCML by blockage of CD59 function is mediated by nAbs, non-nAbs, or both. We found that all nAbs and two non-nAbs (N5-i5 and A32) strongly reacted to three HIV-1 laboratory strains (R5, X4, and R5/ X4), six primary isolates, and provirus-activated ACH-2 cells examined. In contrast, two non-nAbs, 2.2C and N12-i15, reacted weakly and did not react to these targets, respectively. After blockage of CD59 function, the reactive Abs, regardless of their neutralizing activities, significantly enhanced specific ADCML of HIV-1 virions (both laboratory strains and primary isolates) and provirus-activated latently infected cells. The ADMCL efficacy positively correlated with the enzyme-linked immunosorbent assay-reactive intensity of those Abs with their targets. Thus, blockage of RCA function represents a novel approach to restore activities of both nAbs and non-nAbs in triggering ADCML of HIV-1 virions and provirus-activated latently infected cells. IMPORTANCEThere is a renewed interest in the potential role of non-nAbs in the control of HIV-1 infection. Our data, for the first time, demonstrated that blockage of the biological function of RCA members rendered both HIV-1 virions and infected cells sensitive to ADCML mediated by not only nAbs but also non-nAbs. Our results are significant in developing novel immune-based approaches to restore the functions of nAbs and non-nAbs in the circulation of HIV-1-infected individuals to specifically target and clear HIV-1 virions and infected cells. Our data also provide new insights into the mechanisms by which HIV-1 virions and infected cells escape Ab-mediated immunity and could aid in the design and/or development of therapeutic HIV-1 vaccines. In addition, a combination of antiretroviral therapy with RCA blockage, provirus activators, and therapeutic vaccines may represent a novel approach to eliminate HIV-1 reservoirs, i.e., the infected cells harboring replication-competent proviruses and residual viremia.

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“…We sought to assess the contributions of various PBMC subsets, some of which were previously shown to be readily transduced by vaccinia virus, to the production of cytokines and chemokines elicited by the ALVAC, MVA, and NYVAC vectors (13). Human PBMC (n ϭ 2 to 6/group) were isolated as described above and depleted of T cells, monocytes and macrophages (MonoMac), myeloid dendritic cells, or plasmacytoid dendritic cells (pDC) by magnetic separation as previously described (11).…”

mentioning

confidence: 99%

“…Fresh human PBMC (n ϭ 4/group) were isolated by Ficoll-Hypaque density gradient centrifugation (12). In order to include all of the possible cell types shown to be infectible with poxvirus vectors, whole PBMC were infected with the ALVAC, MVA, or NYVAC vector at a multiplicity of infection (MOI) of 10 PFU/cell (13). Culture supernatant was analyzed for cytokine and chemokine levels 24 h postinfection by Luminex assays.…”

mentioning

confidence: 99%

The Canarypox Virus Vector ALVAC Induces Distinct Cytokine Responses Compared to the Vaccinia Virus-Based Vectors MVA and NYVAC in Rhesus Monkeys

Teigler

Phogat

Franchini

et al. 2014

J Virol

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f Despite the growing use of poxvirus vectors as vaccine candidates for multiple pathogens and cancers, their innate stimulatory properties remain poorly characterized. Here we show that the canarypox virus-based vector ALVAC induced distinct systemic proinflammatory and antiviral cytokine and chemokine levels following the vaccination of rhesus monkeys compared to the vaccinia virus-based vectors MVA and NYVAC. These data suggest that there are substantial biological differences among leading poxvirus vaccine vectors that may influence resultant adaptive immune responses following vaccination.

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Primary Human Leukocyte Subsets Differentially Express Vaccinia Virus Receptors Enriched in Lipid Rafts

Cited by 16 publications

References 45 publications

Provirus Activation Plus CD59 Blockage Triggers Antibody-Dependent Complement-Mediated Lysis of Latently HIV-1–Infected Cells

Provirus Activation Plus CD59 Blockage Triggers Antibody-Dependent Complement-Mediated Lysis of Latently HIV-1–Infected Cells

Blockage of CD59 Function Restores Activities of Neutralizing and Nonneutralizing Antibodies in Triggering Antibody-Dependent Complement-Mediated Lysis of HIV-1 Virions and Provirus-Activated Latently Infected Cells

The Canarypox Virus Vector ALVAC Induces Distinct Cytokine Responses Compared to the Vaccinia Virus-Based Vectors MVA and NYVAC in Rhesus Monkeys

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