title: Efficient isolation of novel human monoclonal antibodies with neutralizing activity against HIV-1 from transgenic mice expressing human Ig loci.
abstract: Despite considerable interest in the isolation of mAbs with potent neutralization activity against primary HIV-1 isolates, both for identifying useful targets for vaccine development and for the development of therapeutically useful reagents against HIV-1 infection, a relatively limited number of such reagents have been isolated to date. Human mAbs (hu-mAbs) are preferable to rodent mAbs for treatment of humans, but isolation of hu-mAbs from HIV-infected subjects by standard methods of EBV transformation of B cells or phage display of Ig libraries is inefficient and limited by the inability to control or define the original immunogen. An alternative approach for the isolation of hu-mAbs has been provided by the development of transgenic mice that produce fully hu-mAbs. In this report, we show that immunizing the XenoMouse G2 strain with native recombinant gp120 derived from HIV(SF162) resulted in robust humoral Ab responses against gp120 and allowed the efficient isolation of hybridomas producing specific hu-mAbs directed against multiple regions and epitopes of gp120. hu-mAbs possessing strong neutralizing activity against the autologous HIV(SF162) strain were obtained. The epitopes recognized were located in three previously described neutralization domains, the V2-, V3- and CD4-binding domains, and in a novel neutralization domain, the highly variable C-terminal region of the V1 loop. This is the first report of neutralizing mAbs directed at targets in the V1 region. Furthermore, the V2 and V3 epitopes recognized by neutralizing hu-mAbs were distinct from those of previously described human and rodent mAbs and included an epitope requiring a full length V3 loop peptide for effective presentation. These results further our understanding of neutralization targets for primary, R5 HIV-1 viruses and demonstrate the utility of the XenoMouse system for identifying new and interesting epitopes on HIV-1.
authors.affiliation: Laboratory of Retroviral Biology, Public Health Research Institute, Newark, NJ 07103-3535, USA.
authors.forename: Yuxian
authors.initials: Y
authors.lastname: He
authors.forename: William J
authors.initials: WJ
authors.lastname: Honnen
authors.forename: Chavdar P
authors.initials: CP
authors.lastname: Krachmarov
authors.forename: Michael
authors.initials: M
authors.lastname: Burkhart
authors.forename: Samuel C
authors.initials: SC
authors.lastname: Kayman
authors.forename: Jose
authors.initials: J
authors.lastname: Corvalan
authors.forename: Abraham
authors.initials: A
authors.lastname: Pinter
mesh_terms: D000595:Amino Acid Sequence; D000818:Animals; D019380:Anti-HIV Agents / Q000737:chemistry / Q000302:isolation & purification* / Q000494:pharmacology*; D000910:Antibodies, Heterophile / Q000737:chemistry / Q000235:genetics / Q000302:isolation & purification / Q000494:pharmacology; D000911:Antibodies, Monoclonal / Q000096:biosynthesis / Q000235:genetics / Q000302:isolation & purification* / Q000494:pharmacology*; D000918:Antibody Specificity / Q000235:genetics; D001667:Binding, Competitive / Q000235:genetics / Q000276:immunology; D017124:Conserved Sequence / Q000276:immunology; D018604:Epitope Mapping; D000939:Epitopes / Q000737:chemistry / Q000276:immunology; D005786:Gene Expression Regulation / Q000276:immunology; D005803:Genes, Immunoglobulin; D005819:Genetic Markers / Q000276:immunology; D015483:HIV Antibodies / Q000096:biosynthesis / Q000235:genetics / Q000302:isolation & purification* / Q000494:pharmacology*; D015699:HIV Envelope Protein gp120 / Q000276:immunology; D015497:HIV-1 / Q000276:immunology*; D006801:Humans; D006825:Hybridomas; D007135:Immunoglobulin Variable Region / Q000737:chemistry / Q000235:genetics; D051379:Mice; D008822:Mice, Transgenic; D008969:Molecular Sequence Data; D009500:Neutralization Tests / Q000379:methods; D017434:Protein Structure, Tertiary / Q000235:genetics
chemical_list: D019380:Anti-HIV Agents; D000910:Antibodies, Heterophile; D000911:Antibodies, Monoclonal; D000939:Epitopes; D005819:Genetic Markers; D015483:HIV Antibodies; D015699:HIV Envelope Protein gp120; D007135:Immunoglobulin Variable Region