title: Improvement in affinity and HIV-1 neutralization by somatic mutation in the heavy chain first complementarity-determining region of antibodies triggered by HIV-1 infection.
abstract: We assessed the impact of somatic hypermutation in the framework region 1 (FR1) and complementarity-determining region 1 (CDR1) of three clonally-related heavy chains from the human monovalent antigen-binding fragments Fab S19, S8 and S20 on gp120 binding and HIV-1 neutralization capacity. Nucleotide changes were introduced in the heavy chains to revert single and multiple amino acid residues, and two Fab libraries were constructed with the same light chain to express equivalent amounts of parental and reverted phage Fab. We studied the contribution of each amino acid replacement to antigen binding by calculating the frequency of phage Fab retrieval after competitive library selection on gp120. Whereas mutations in FR1 had no effect on antigen binding, somatic replacements in the CDR1 of the heavy chain (HCDR1) appeared to produce significant changes. In S19 HCDR1, somatic mutation of residue 32 reduced gp120 binding. In Fab S20, the Arg(30) and Asp(31) somatically replaced residues in HCDR1 improved antigen binding. Both of these residues are necessary to increase Fab binding to gp120; reversion of either residue alone results in a decrease in binding. The impact of these two replacements was confirmed by the greater neutralization capacity of S20 compared to the other Fab. Molecular modeling of S20 HCDR1 suggests that Arg(30) and Asp(31) are the main interaction sites for gp120, increasing antibody affinity and promoting the enhanced neutralization ability of S20. These findings are consistent with a gp120-driven process, supporting a role for affinity maturation and intraclonal evolution of HIV-1 neutralizing antibodies.
authors.affiliation: Department of Immunology and Oncology, UAM Campus de Cantoblanco, Madrid, Spain. jltoran@cnb.uam.es
authors.forename: J L
authors.initials: JL
authors.lastname: Torán
authors.forename: L
authors.initials: L
authors.lastname: Sánchez-Pulido
authors.forename: L
authors.initials: L
authors.lastname: Kremer
authors.forename: G
authors.initials: G
authors.lastname: del Real
authors.forename: A
authors.initials: A
authors.lastname: Valencia
authors.forename: C
authors.initials: C
authors.lastname: Martínez-A
mesh_terms: D000595:Amino Acid Sequence; D000915:Antibody Affinity; D001435:Bacteriophages / Q000235:genetics; D001483:Base Sequence; D022801:Complementarity Determining Regions / Q000276:immunology*; D015483:HIV Antibodies / Q000276:immunology*; D015699:HIV Envelope Protein gp120 / Q000276:immunology*; D015497:HIV-1 / Q000276:immunology*; D006801:Humans; D007140:Immunoglobulin Fab Fragments / Q000276:immunology; D008969:Molecular Sequence Data; D009154:Mutation; D009500:Neutralization Tests; D013329:Structure-Activity Relationship
chemical_list: D022801:Complementarity Determining Regions; D015483:HIV Antibodies; D015699:HIV Envelope Protein gp120; D007140:Immunoglobulin Fab Fragments