Journal: The Journal of Infectious Diseases

Article Title: Pathological Role of Anti-CD4 Antibodies in HIV-Infected Immunologic Nonresponders Receiving Virus-Suppressive Antiretroviral Therapy

doi: 10.1093/infdis/jix223

Figure Lengend Snippet: Clinical Characteristics of Study Participants

Article Snippet: We thank the AIDS Reagent Program, Division of AIDS, National Institute of Allergy and Infectious Diseases, National Institutes of Health, for providing human soluble CD4 recombinant protein (Progenics); and Dr Paul Parren (Genmab), for providing human monoclonal anti-CD4 antibody (zanolimumab).

Techniques:

Journal: The Journal of Infectious Diseases

Article Title: Pathological Role of Anti-CD4 Antibodies in HIV-Infected Immunologic Nonresponders Receiving Virus-Suppressive Antiretroviral Therapy

doi: 10.1093/infdis/jix223

Figure Lengend Snippet: Increased plasma anti-CD4 immunoglobulin G (IgG) levels in aviremic antiretroviral therapy (ART)–recipient immunologic nonresponders. A, Median absolute numbers of peripheral CD4+ and CD8+ T cells, assessed by flow cytometry, in healthy controls, responders, and nonresponders. B, Median plasma levels of anti-CD4 IgG in healthy controls, responders, nonresponders, and long-term nonprogressors (LTNPs). C and D, Correlations between plasma levels of anti-CD4 IgG and peripheral CD4+ T-cell counts in all ART-recipient aviremic human immunodeficiency virus (HIV)–infected subjects (C) and healthy controls (D). E, Median plasma levels of soluble CD4 (sCD4) antigen in healthy controls, responders, and nonresponders. Statistical analyses were performed using the Mann-Whitney U test (unpaired).

Article Snippet: We thank the AIDS Reagent Program, Division of AIDS, National Institute of Allergy and Infectious Diseases, National Institutes of Health, for providing human soluble CD4 recombinant protein (Progenics); and Dr Paul Parren (Genmab), for providing human monoclonal anti-CD4 antibody (zanolimumab).

Techniques: Flow Cytometry, Infection, MANN-WHITNEY

Journal: The Journal of Infectious Diseases

Article Title: Pathological Role of Anti-CD4 Antibodies in HIV-Infected Immunologic Nonresponders Receiving Virus-Suppressive Antiretroviral Therapy

doi: 10.1093/infdis/jix223

Figure Lengend Snippet: Anti-CD4 antibody-dependent natural killer (NK) cell activation. CD4+ T cells were cultured with purified anti-CD4 immunoglobulin G (IgG) from nonresponders or control antibodies and cocultured with NK cells at a ratio of 1:1. Intracellular CD107a and interferon γ (IFN-γ) expression in NK cells was analyzed by flow cytometry. A, Dot plots from a representative donor show CD107a and IFN-γ expression in NK cells in response to different concentrations of anti-CD4 antibodies from nonresponders. B, Percentages of NK cells expressing IFN-γ and CD107a in response to different concentrations of anti-CD4 IgG from 5 different nonresponders in vitro. Analyses were performed by the Friedman paired nonparametric test. C, Median percentages of NK cells expressing IFN-γ and CD107a in a mixed culture of CD4+ T cells and NK cells in the presence of anti-CD4 IgG from nonresponders, anti-CD4 IgG from nonresponders pretreated with sCD4 (control 1), anti-CD4 IgG–depleted total IgG from nonresponders (control 2), and zanolimumab (positive control) at 5 μg/mL in vitro. Analyses were performed using the Mann-Whitney U test (unpaired). D, Median percentages of NK cells expressing CD107a in healthy controls, responders, and nonresponders ex vivo. Analyses were performed using the Mann-Whitney U test (unpaired). FSC, forward scatter; SSC, side scatter.

Article Snippet: We thank the AIDS Reagent Program, Division of AIDS, National Institute of Allergy and Infectious Diseases, National Institutes of Health, for providing human soluble CD4 recombinant protein (Progenics); and Dr Paul Parren (Genmab), for providing human monoclonal anti-CD4 antibody (zanolimumab).

Techniques: Activation Assay, Cell Culture, Purification, Expressing, Flow Cytometry, In Vitro, Positive Control, MANN-WHITNEY, Ex Vivo

Journal: The Journal of Infectious Diseases

Article Title: Pathological Role of Anti-CD4 Antibodies in HIV-Infected Immunologic Nonresponders Receiving Virus-Suppressive Antiretroviral Therapy

doi: 10.1093/infdis/jix223

Figure Lengend Snippet: Anti-CD4 antibody dependent natural killer (NK) cell–mediated cytolysis to primary CD4+ T cells. CD4+ T cells were cultured with purified anti-CD4 immunoglobulin G (IgG) and control antibodies and cocultured with NK cells at a ratio of 1:3 in vitro, and the CD4+ T-cell cytolysis percentage was analyzed by flow cytometry. A, Dot plots from 1 representative donor were showing CD4+ T-cell frequencies and annexin V binding in response to different concentrations of anti-CD4 IgG from nonresponders. B, Percentages of CD4+ T cells undergoing cytolysis in response to different concentrations of anti-CD4 IgG from 5 different nonresponders through antibody-dependent cell-mediated cytotoxicity (ADCC). Analyses were performed by the Friedman paired nonparametric test. C, Median percentages of CD4+ T cells undergoing cytolysis in response to anti-CD4 IgG from nonresponders and controls at 5 μg/mL in vitro. Analyses were performed using the Mann-Whitney U test (unpaired). D, Percentages of CD4+ T cells undergoing apoptosis in response to different concentrations of anti-CD4 IgG from 5 different nonresponders through ADCC. Analyses were performed by the Friedman paired nonparametric test. E, Percentages of CD4+ T cells undergoing apoptosis in response to anti-CD4 IgG from nonresponders and controls at 5 μg/mL in vitro. Analyses were performed using the Mann-Whitney U test (unpaired). SSC, side scatter.

Article Snippet: We thank the AIDS Reagent Program, Division of AIDS, National Institute of Allergy and Infectious Diseases, National Institutes of Health, for providing human soluble CD4 recombinant protein (Progenics); and Dr Paul Parren (Genmab), for providing human monoclonal anti-CD4 antibody (zanolimumab).

Techniques: Cell Culture, Purification, In Vitro, Flow Cytometry, Binding Assay, MANN-WHITNEY

Journal: The Journal of Infectious Diseases

Article Title: Pathological Role of Anti-CD4 Antibodies in HIV-Infected Immunologic Nonresponders Receiving Virus-Suppressive Antiretroviral Therapy

doi: 10.1093/infdis/jix223

Figure Lengend Snippet: CD4+ T-cell activation in response to purified anti-CD4 immunoglobulin G (IgG) from immunologic nonresponders. A, Overlapped dot plots (1 representative donor) and the median CD4 mean fluorescence intensity (MFI) expression on CD4+ T cells treated with anti-CD4 IgG (red) and anti-CD4 IgG–depleted total IgG (blue) in vitro. B and C, Median CD4 MFI expression on naive CD4+ T cells (B) and memory CD4+ T cells (C) in healthy controls, responders, and nonresponders ex vivo. D, Overlapped dot plots (1 representative donor) and median percentages of CD38 and HLA-DR coexpression on memory CD4+ T cells treated with anti-CD4 IgG (red) and anti-CD4 IgG–depleted total IgG in vitro. E, Median percentages of CD38 and HLA-DR coexpression on memory CD4+ T cells in healthy controls, responders, and nonresponders ex vivo. Analyses were performed using the Mann-Whitney U test (unpaired). SSC, side scatter.

Article Snippet: We thank the AIDS Reagent Program, Division of AIDS, National Institute of Allergy and Infectious Diseases, National Institutes of Health, for providing human soluble CD4 recombinant protein (Progenics); and Dr Paul Parren (Genmab), for providing human monoclonal anti-CD4 antibody (zanolimumab).

Techniques: Activation Assay, Purification, Fluorescence, Expressing, In Vitro, Ex Vivo, MANN-WHITNEY

Journal: The Journal of Infectious Diseases

Article Title: Pathological Role of Anti-CD4 Antibodies in HIV-Infected Immunologic Nonresponders Receiving Virus-Suppressive Antiretroviral Therapy

doi: 10.1093/infdis/jix223

Figure Lengend Snippet: Memory and naive CD4+ T cells. A, Comparison of apoptotic induction between naive CD4+ and memory CD4+ T cells by purified anti-CD4 immunoglobulin G (IgG) from plasma of 5 different nonresponders in antibody-dependent cell-mediated cytotoxicity assays in vitro. Data are median fold change (interquartile range) from anti-CD4 IgG to anti-CD4 IgG–depleted total IgG. B, Median ratio of memory CD4+ T cells to naive CD4+ T cells in healthy controls, responders, and nonresponders ex vivo. C, Median absolute counts of memory CD4+ and naive CD4+ T cells in healthy controls, responders, and nonresponders ex vivo. Analyses were performed using the Mann-Whitney U test (unpaired).

Article Snippet: We thank the AIDS Reagent Program, Division of AIDS, National Institute of Allergy and Infectious Diseases, National Institutes of Health, for providing human soluble CD4 recombinant protein (Progenics); and Dr Paul Parren (Genmab), for providing human monoclonal anti-CD4 antibody (zanolimumab).

Techniques: Purification, In Vitro, Ex Vivo, MANN-WHITNEY

Journal: AIDS Research and Human Retroviruses

Article Title: The Potent Human Immunodeficiency Virus Type 1 (HIV-1) Entry Inhibitor HR212 Blocks Formation of the Envelope Glycoprotein gp41 Six-Helix Bundle

doi: 10.1089/aid.2012.0059

Figure Lengend Snippet: (A) Cell–cell fusion assays examining the inhibitory effects of HR212 on HIV-1-mediated cell–cell fusion between H9/HTLV-IIIB and MT-2 cells. (B) Inhibition of gp120 binding to CD4 by HR212. The CD4-based ELISA was carried out in the presence of HR212 (left bar) or the CD4 antibody (RPA-T4) (right bar). (C) Inhibition of the anti-CXCR4 mAb 12G5 binding to CXCR4. Inhibition of HR212 (20 μM, left bar) and the positive control AMD3100 (10 μM, right bar) on the binding of the anti-CXCR4 antibody 12G5 to U373-MAGI-CXCR4CEM cells was determined using a cell-based enzyme-linked immunosorbent assay (ELISA). (D) Inhibition of gp120–CD4 complex binding to CCR5. Inhibition of HR212 (20 μM, left bar) and the CCR5 antagonist maraviroc (1 μM, right bar) on the binding of the gp120–CD4 complex to cells expressing CCR5 was determined using a cell-based ELISA. Each sample was tested in triplicate in two independent experiments, and the data are presented as the means±the standard deviations.

Article Snippet: A 1:1 mix of soluble human CD4 (eENZYME, Gaithersburg, MD) (2 μg/ml) and HIV-1 gp120 (Bal/Clade B) (eENZYME, Gaithersburg, MD) (2 μg/ml) was incubated at room temperature for 15 min prior to its addition to the plate in the presence of HR212 (20 μM) or maraviroc (Pfizer) (1 μM).

Techniques: Inhibition, Binding Assay, Enzyme-linked Immunosorbent Assay, Positive Control, Expressing, In-Cell ELISA