title: Cross-reactive HIV-1 neutralizing monoclonal antibodies selected by screening of an immune human phage library against an envelope glycoprotein (gp140) isolated from a patient (R2) with broadly HIV-1 neutralizing antibodies.
abstract: Elicitation of broadly cross-reactive neutralizing antibodies (bcnAbs) in HIV infections is rare. To test the hypothesis that such antibodies could be elicited by HIV envelope glycoproteins (Envs) with unusual immunogenic properties and to identify novel bcnAbs, we used a soluble Env ectodomain (gp140) from a donor (R2) with high level of bcnAbs as an antigen for panning of an immune phage-displayed antibody library. The panning with the R2 Env resulted in significantly higher number of cross-reactive antibody clones than by using Envs from two other isolates (89.6 and IIIB). Two of the identified human monoclonal antibodies (hmAbs), m22 and m24, had sequences, neutralizing and binding activities similar or identical to those of the gp120-specific bcnAbs m18 and m14. The use of the R2 Env but not other Envs for panning resulted in the identification of a novel gp41-specific hmAb, m46. For several of the tested HIV-1 primary isolates its potency on molar basis was comparable to that of T20. It inhibited entry of primary isolates from different clades with an increased activity for cell lines with low CCR5 surface concentrations. The m46 neutralizing activity against a panel of clade C isolates was significantly higher in an assay based on peripheral blood mononuclear cells (4 out of 5 isolates were neutralized with an IC(50) in the range from 1.5 to 25 microg/ml) than in an assay based on a cell line with relatively high concentration of cell-surface-associated CCR5. In contrast to 2F5 and Z13, this antibody did not bind to denatured gp140 and gp41-derived peptides indicating a conformational nature of its epitope. It bound to a 5-helix bundle but not to N-heptad repeat coiled coils and a 6-helix bundle construct indicating contribution of both gp41 heptad repeats to its epitope and to a possible mechanism of neutralization. These results indicate that the R2 Env may contain unique exposed conserved epitopes that could contribute to its ability to elicit broadly cross-reactive antibodies in animals and humans; the newly identified antibodies may help in the development of novel vaccine immunogens and therapeutics.
authors.affiliation: Protein Interactions Group, CCRNP, NCI-Frederick, NIH, Frederick, MD 21702, USA.
authors.forename: Vidita
authors.initials: V
authors.lastname: Choudhry
authors.forename: Mei-Yun
authors.initials: MY
authors.lastname: Zhang
authors.forename: Igor A
authors.initials: IA
authors.lastname: Sidorov
authors.forename: John M
authors.initials: JM
authors.lastname: Louis
authors.forename: Ilia
authors.initials: I
authors.lastname: Harris
authors.forename: Antony S
authors.initials: AS
authors.lastname: Dimitrov
authors.forename: Peter
authors.initials: P
authors.lastname: Bouma
authors.forename: Fatim
authors.initials: F
authors.lastname: Cham
authors.forename: Anil
authors.initials: A
authors.lastname: Choudhary
authors.forename: Susanna M
authors.initials: SM
authors.lastname: Rybak
authors.forename: Timothy
authors.initials: T
authors.lastname: Fouts
authors.forename: David C
authors.initials: DC
authors.lastname: Montefiori
authors.forename: Christopher C
authors.initials: CC
authors.lastname: Broder
authors.forename: Gerald V
authors.initials: GV
authors.lastname: Quinnan
authors.forename: Dimiter S
authors.initials: DS
authors.lastname: Dimitrov
mesh_terms: D000911:Antibodies, Monoclonal / Q000276:immunology* / Q000302:isolation & purification; D001667:Binding, Competitive; D015153:Blotting, Western; D002460:Cell Line; D003429:Cross Reactions / Q000276:immunology*; D004797:Enzyme-Linked Immunosorbent Assay; D015686:Gene Products, env / Q000276:immunology*; D015483:HIV Antibodies / Q000276:immunology*; D015699:HIV Envelope Protein gp120 / Q000276:immunology; D015497:HIV-1 / Q000145:classification / Q000276:immunology*; D006801:Humans; D007140:Immunoglobulin Fab Fragments / Q000235:genetics / Q000276:immunology; D009500:Neutralization Tests; D019151:Peptide Library; D054299:env Gene Products, Human Immunodeficiency Virus
chemical_list: D000911:Antibodies, Monoclonal; D015686:Gene Products, env; D015483:HIV Antibodies; D015699:HIV Envelope Protein gp120; D007140:Immunoglobulin Fab Fragments; D019151:Peptide Library; D054299:env Gene Products, Human Immunodeficiency Virus; C097729:gp140 envelope protein, Human immunodeficiency virus 1