The antigen-binding characteristics of mAbs derived from in vivo priming of avian B cells.
In most vertebrates, a primary antibody repertoire is created through the recombination of a diverse set of Ig variable (V), diversity (D), and joining (J) gene segments. In contrast, an avian immune repertoire is generated by gene conversion of rearranged Ig genes during B cell development within the bursa of Fabricius, a lymphoid organ unique to birds. To investigate the properties of antigen-specific Igs created through the process of gene conversion, we have developed a system for the production of avian-derived mAbs. This system was used to produce multiple antibodies after a single immunization with a conserved peptide from the human cystic fibrosis transmembrane conductance regulator gene. Each antibody isolated was found to have arisen independently through a distinct series of gene conversion events. These primary antibodies displayed evidence of diversity in all of the complementarity determining regions of both heavy and light chains, and both the heavy and the light chains contributed to antigen specificity. In the light chains, diversity could be attributed to gene conversion events. The measured affinity constants of two of the antibodies were between 10(8) and 10(9) M-1, and the antibodies were functional in quantitative ELISA as well as immunohistochemical studies of cystic fibrosis transmembrane conductance regulator expression. These data demonstrate that antigen-specific antibodies produced by Ig gene conversion display both high affinity and specificity. In addition, the methods developed here provide the description of a system for the production of mAbs derived from a nonmammalian species.
Michael
N
N
Gwen Knapp Center for Lupus and Immunology Research, The University of Chicago, Chicago, IL 60637, USA.
Accavitti
M A
MA
Masteller
E
E
Thompson
C B
CB
D000595:Amino Acid Sequence; D000818:Animals; D000911:Antibodies, Monoclonal / Q000737:chemistry / Q000276:immunology*; D000916:Antibody Diversity / Q000235:genetics / Q000276:immunology; D000918:Antibody Specificity; D001402:B-Lymphocytes / Q000276:immunology*; D002645:Chickens; D019005:Cystic Fibrosis Transmembrane Conductance Regulator / Q000737:chemistry / Q000235:genetics / Q000276:immunology; D005091:Exons; D015322:Gene Rearrangement, B-Lymphocyte; D006801:Humans; D007143:Immunoglobulin Heavy Chains / Q000235:genetics / Q000276:immunology; D007144:Immunoglobulin J-Chains / Q000235:genetics / Q000276:immunology; D007147:Immunoglobulin Light Chains / Q000235:genetics / Q000276:immunology; D007135:Immunoglobulin Variable Region / Q000235:genetics / Q000276:immunology; D008969:Molecular Sequence Data; D011993:Recombinant Fusion Proteins / Q000737:chemistry / Q000276:immunology
D000911:Antibodies, Monoclonal; C505032:CFTR protein, human; D007143:Immunoglobulin Heavy Chains; D007144:Immunoglobulin J-Chains; D007147:Immunoglobulin Light Chains; D007135:Immunoglobulin Variable Region; D011993:Recombinant Fusion Proteins; D019005:Cystic Fibrosis Transmembrane Conductance Regulator