title: Further characterization of an antigenic site of HIV-1 gp120 recognized by virus neutralizing human monoclonal antibodies.
abstract: OBJECTIVE
  The aim of this study is to characterize antigenic sites on HIV-1 gp120 which may be important for the development of active and passive immunization strategies against HIV-1 infection.
  
  
  METHODS
  Two HIV-1-seropositive individuals were selected from the Amsterdam cohort and Epstein-Barr virus (EBV)-transformed B cells were generated from their peripheral blood mononuclear cells, which produce HIV-1-specific human monoclonal antibodies (HuMAb).
  
  
  METHODS
  HuMAb were generated and selected based on their reactivities with native gp120. Reactivity with HIV-1 strains from phylogenetically different subfamilies was determined by immunostaining and virus neutralization assays. Specificity for the CD4-binding site was tested by an inhibition enzyme-linked immunosorbent assay and amino acids (aa) involved in the binding of the HuMAb were identified with a set of gp120 molecules with single aa substitutions.
  
  
  RESULTS
  Three HuMAb (GP13, GP44, GP68) were generated, all recognizing a conserved conformation dependent epitope within, or topographically near, the CD4-binding site of gp120. HuMAb GP13 and GP68 neutralized a broad range of HIV-1 strains from phylogenetically different subfamilies, whereas HuMAb GP44 exhibited a more restricted pattern of neutralizing activity. The patterns of gp120 aa involved in their binding were unique for each of these HuMAb.
  
  
  CONCLUSIONS
  The pattern of reactivities of these three HIV-1-neutralizing HuMAb developed in these studies is similar to, but distinct from other human and rodent MAb that recognize this antigenic site of HIV-1 gp120.
authors.affiliation: Laboratory of Immunobiology, National Institute of Public Health and Environmental Protection, Bilthoven, The Netherlands.
authors.forename: M
authors.initials: M
authors.lastname: Schutten
authors.forename: A
authors.initials: A
authors.lastname: McKnight
authors.forename: R C
authors.initials: RC
authors.lastname: Huisman
authors.forename: M
authors.initials: M
authors.lastname: Thali
authors.forename: J A
authors.initials: JA
authors.lastname: McKeating
authors.forename: J
authors.initials: J
authors.lastname: Sodroski
authors.forename: J
authors.initials: J
authors.lastname: Goudsmit
authors.forename: A D
authors.initials: AD
authors.lastname: Osterhaus
mesh_terms: D000328:Adult; D000911:Antibodies, Monoclonal / Q000276:immunology*; D000914:Antibodies, Viral / Q000276:immunology*; D000918:Antibody Specificity; D001402:B-Lymphocytes / Q000382:microbiology; D002999:Clone Cells / Q000382:microbiology; D004797:Enzyme-Linked Immunosorbent Assay; D000939:Epitopes / Q000276:immunology*; D015699:HIV Envelope Protein gp120 / Q000276:immunology*; D015497:HIV-1 / Q000276:immunology*; D006801:Humans; D007150:Immunohistochemistry; D008297:Male; D009426:Netherlands; D009500:Neutralization Tests
chemical_list: D000911:Antibodies, Monoclonal; D000914:Antibodies, Viral; D000939:Epitopes; D015699:HIV Envelope Protein gp120