A novel enzyme-linked immunosorbent assay (ELISA) for the detection of antibodies to HIV-1 envelope glycoproteins based on immobilization of rival glycoproteins in microtiter wells coated with concanavalin A

Robinson, James E.; Holton, Debra; Liu, James H.; McMURDO, Hal; Murciano, A. Attias de; Gohd, Robert S.

doi:10.1016/0022-1759(90)90399-g

Cited by 56 publications

(33 citation statements)

References 21 publications

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“…The source of native J62 gp 120 was detergent-treated cellfree culture fluid from a chronically infected continuous T cell line. We also established in earlier studies that 2.3a and l.5e showed equivalent reactivity with both LAV rgpl20 and native J62 gpl20 in an ELISA in which gpl20 was immobilized in Con A-coated wells (8). Statistics.…”

Section: Methodssupporting

confidence: 58%

See 1 more Smart Citation

Distinction of human immunodeficiency virus type 1 neutralization and infection enhancement by human monoclonal antibodies to glycoprotein 120.

Takeda¹,

Robinson²,

Ho³

et al. 1992

J. Clin. Invest.

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There is increasing evidence that sera from HIV-1-infected individuals contain antibodies that enhance infection by HIV-1 in vitro. Previous work has demonstrated that complement receptors on T lymphoid cells and Fc receptors for IgG (Fc'yR) on monocytic cells are required for enhanced infection by antibody-complexed HIV-1. Characterization of such infection-enhancing antibodies is essential because immunogenic epitopes which induce enhancing antibodies should be excluded from HIV-1 vaccines. This study was conducted to identify enhancing antibodies involved in Fc R-mediated enhancement of HIV-1 infection employing IgG human monoclonal antibodies (HMAbs) reactive against gpl20 of HIV-1, which were produced by B cell lines derived from an HIV-1-infected individual. A potent neutralizing HMAb N70-1.5e did not enhance infection by HIV-1 (IIIB and MN strains), whereas HMAb N70-2.3a mediated enhancement of HIV-1 infection, but had little neutralizing activity. A competition radio immunoassay demonstrated that the two antibodies bind to distinct epitopes. These results indicated that enhancing and neutralizing antibodies can be induced by different epitopes on gpl20, suggesting the potential for development of safe vaccines against HIV-1 by exclusion of immunogenic epitopes for enhancing antibodies. We made attempts to identify the epitope on gpl20 that is recognized by the enhancing antibody N70-23a by using recombinant HIV-1 proteins and found that the antibody binds to a conformational site of nonvariable sequences in the carboxyl half (aa 272-509) of gpl20. (J. Clin. Invest. 1992. 89:1952-1957

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Section: Methodssupporting

confidence: 58%

“…These strains were grown in H9 cells as acutely infected cultures. A local strain, HIV-I J62 (8), that was isolated from the peripheral blood mononuclear cells of an HIV-l-infected patient by cocultivation with mitogen-stimulated T cells from seronegative blood, was used to prepare native gp 120 antigen for binding assays.…”

Section: Methodsmentioning

confidence: 99%

Distinction of human immunodeficiency virus type 1 neutralization and infection enhancement by human monoclonal antibodies to glycoprotein 120.

Takeda¹,

Robinson²,

Ho³

et al. 1992

J. Clin. Invest.

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“…The production of hMAbs by Epstein-Barr virus (EBV) transformation of B cells has been described previously (30,(36)(37)(38). Using this method, transformed clonal B cell lines were produced for hMAbs 4.8A (30) (39).…”

Section: Methodsmentioning

confidence: 99%

“…Bcell cultures were screened for antibody production using ELISA as described previously (30,36). Briefly, 96-well plates (Costar; Corning, Corning, NY) were coated with ConA at 25 g/ml in 0.01 M HEPES (Gibco) for 1 h. The wells were washed (PBS containing 0.1% [vol/vol] Tween 20), and Triton X-100-solubilized DENV produced in serum-free medium was incubated for 1 h. All steps in this ELISA were performed at room temperature.…”

Section: Methodsmentioning

confidence: 99%

Mechanistic Study of Broadly Neutralizing Human Monoclonal Antibodies against Dengue Virus That Target the Fusion Loop

Costin

Зайцева

Kahle

et al. 2013

J Virol

Self Cite

117

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There are no available vaccines for dengue, the most important mosquito-transmitted viral disease. Mechanistic studies with anti-dengue virus (DENV) human monoclonal antibodies (hMAbs) provide a rational approach to identify and characterize neutralizing epitopes on DENV structural proteins that can serve to inform vaccine strategies. Here, we report a class of hMAbs that is likely to be an important determinant in the human humoral response to DENV infection. In this study, we identified and characterized three broadly neutralizing anti-DENV hMAbs: 4.8A, D11C, and 1.6D. These antibodies were isolated from three different convalescent patients with distinct histories of DENV infection yet demonstrated remarkable similarities. All three hMAbs recognized the E glycoprotein with high affinity, neutralized all four serotypes of DENV, and mediated antibody-dependent enhancement of infection in Fc receptor-bearing cells at subneutralizing concentrations. The neutralization activities of these hMAbs correlated with a strong inhibition of virus-liposome and intracellular fusion, not virus-cell binding. We mapped epitopes of these antibodies to the highly conserved fusion loop region of E domain II. Mutations at fusion loop residues W101, L107, and/or G109 significantly reduced the binding of the hMAbs to E protein. The results show that hMAbs directed against the highly conserved E protein fusion loop block viral entry downstream of virus-cell binding by inhibiting E protein-mediated fusion. Characterization of hMAbs targeting this region may provide new insights into DENV vaccine and therapeutic strategies.

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“…Anti-gp160 antibodies were quantitated as previously described (8,31). Briefly, 96-well ELISA plates (Immulon II; Dynex Technologies) were coated with 0.5 g of concavalin A (ConA; Sigma) per well.…”

Section: Methodsmentioning

confidence: 99%

Cytokine Expression, Natural Killer Cell Activation, and Phenotypic Changes in Lymphoid Cells from Rhesus Macaques during Acute Infection with Pathogenic Simian Immunodeficiency Virus

Giavedoni

Velasquillo²,

Parodi³

et al. 2000

J Virol

120

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We studied the innate and adaptive immune system of rhesus macaques infected with the virulent simian immunodeficiency virus isolate SIVmac251 by evaluating natural killer (NK) cell activity, cytokine levels in plasma, humoral and virological parameters, and changes in the activation markers CD25 (interleukin 2R The immune system of higher vertebrates consists of innate and adaptive components. Innate immunity exhibits immediate recognition and response without prior sensitization. Cells of the innate immune system (i.e., monocytes/macrophages, natural killer [NK] cells, and polymorphonuclear leukocytes) recognize pathogen-associated molecular patterns and activate events such as phagocytosis, induction of the synthesis of antimicrobial peptides, expression of inflammatory and effector cytokines and chemokines, induction of nitric oxide synthase in macrophages, and expression of costimulatory molecules on antigen-presenting cells. The adaptive immune system uses somatically generated antigen receptors that are clonally distributed on T and B lymphocytes. Generally, adaptive immune recognition in the absence of innate immune recognition results in inactivation of lymphocytes that express receptors involved in the identification events (20). Thus, innate immune responses have critical consequences in adaptive immune responses.[Little is known of the contribution of the innate immune system during infection with the human immunodeficiency virus (HIV). Based on similarities of biologic and genetic features, simian immunodeficiency virus (SIV) infection of rhesus macaques provides the best animal model of HIV infection and AIDS. Accordingly, this animal model is critical for the elucidation of mechanisms of pathogenesis and for the development of vaccines and antiviral therapies (12). As with almost all viral infections, the innate immune system is thought to be the first component of the immune system that recognizes SIV infection. However, few studies have methodically analyzed the changes induced in cell phenotype and cytokine levels by SIV infection. Recent studies have demonstrated that SIV infection results in a generalized increase in lymphocyte turnover (23) and that the primary site for viral replication is activated memory CD4 ϩ T cells that are present in the intestinal lamina propia (46). Although cellular changes are not that dramatic at this early stage in peripheral lymphoid tissue, peripheral blood (PB) and lymph nodes (LN) still reflect the pathologic changes induced by the viral infection and are readily available for longitudinal studies.To analyze changes in the activation state of cells from the innate and adaptive immune system after SIV infection, we evaluated NK activity, cytokine levels in plasma, and changes in activation markers on lymphoid cells of rhesus macaques after infection with pathogenic SIVmac251. We found the sequential appearance in plasma of interferon-␣/␤ (IFN-␣/␤) interleukin-18 (IL-18) and IL-12, whereas IL-4, IFN-␥ and granulocyte-macrophage colony-stimulating factor (GM-CS...

show abstract

A novel enzyme-linked immunosorbent assay (ELISA) for the detection of antibodies to HIV-1 envelope glycoproteins based on immobilization of rival glycoproteins in microtiter wells coated with concanavalin A

Cited by 56 publications

References 21 publications

Distinction of human immunodeficiency virus type 1 neutralization and infection enhancement by human monoclonal antibodies to glycoprotein 120.

Distinction of human immunodeficiency virus type 1 neutralization and infection enhancement by human monoclonal antibodies to glycoprotein 120.

Mechanistic Study of Broadly Neutralizing Human Monoclonal Antibodies against Dengue Virus That Target the Fusion Loop

Cytokine Expression, Natural Killer Cell Activation, and Phenotypic Changes in Lymphoid Cells from Rhesus Macaques during Acute Infection with Pathogenic Simian Immunodeficiency Virus

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