The importance of antibody isotype in HIV-1 virus capture assay and in TZM-bl neutralization

Viral Immunol. 2010 Dec;23(6):627-32. doi: 10.1089/vim.2010.0061.

Abstract

The binding of murine IgM mAbs to five different clades of HIV-1 was examined using a modified ELISA-based virus capture assay. Two murine multispecific IgM mAbs that exhibit both lipid and gp41 epitope specificities, and one murine IgM mAb that exhibits lipid-binding specificity, were utilized. The binding of the IgG and the IgM isotypes of human mAb 2F5 to clades A through AE were also evaluated. The binding of 2F5 to HIV-1 was dependent upon the antibody isotype. Monoclonal IgM antibodies bound significantly lower amounts of HIV-1 than the corresponding IgG isotype. Although murine IgM mAbs bound HIV-1 to varying degrees in the virus capture assay, they failed to neutralize HIV-1 in a TZM-bl pseudovirus assay. In contrast, 2F5-IgM mAb bound certain HIV-1 isolates, and also neutralized them, although not as efficiently as the 2F5-IgG isotype. Studies on the relationship between virus binding and neutralization in a TZM-bl pseudovirus assay indicated that in most cases, mAbs that exhibited neutralization also bound the virus.

Publication types

  • Research Support, Non-U.S. Gov't
  • Research Support, U.S. Gov't, Non-P.H.S.

MeSH terms

  • Animals
  • Antibodies, Neutralizing / immunology*
  • Biological Assay / methods
  • Enzyme-Linked Immunosorbent Assay / methods
  • Epitopes / immunology
  • HEK293 Cells
  • HIV Antibodies / classification*
  • HIV Antibodies / immunology*
  • HIV Core Protein p24 / immunology
  • HIV Envelope Protein gp41 / immunology
  • HIV-1 / classification
  • HIV-1 / immunology*
  • Humans
  • Immunoglobulin G / immunology
  • Immunoglobulin Isotypes / classification*
  • Immunoglobulin Isotypes / immunology*
  • Immunoglobulin M / immunology
  • Mice
  • Neutralization Tests / methods*
  • Protein Binding
  • Viral Tropism / physiology

Substances

  • Antibodies, Neutralizing
  • Epitopes
  • HIV Antibodies
  • HIV Core Protein p24
  • HIV Envelope Protein gp41
  • Immunoglobulin G
  • Immunoglobulin Isotypes
  • Immunoglobulin M