A systematic approach to defining the germline gene counterparts of a mutated autoantibody from a patient with rheumatoid arthritis.
OBJECTIVE
To define the germline counterparts of potentially highly mutated autoantibodies in disease states.


METHODS
We developed a systematic approach by first characterizing a rearranged Ig gene and its upstream flank, and then designing suitable primers to amplify specifically the putative germline counterpart.


RESULTS
We identified and characterized the germline counterpart of a rheumatoid factor heavy chain variable region.


CONCLUSIONS
We showed unequivocally that the heavy chain of a rheumatoid factor, derived from synovial tissue, has 4 replacement mutations from the corresponding germline gene. The technique allows quick assessment of the degree of somatic mutation in many autoantibodies, and thus can help to elucidate the underlying mechanisms for the induction and sustained production of such antibodies in patients.
Soto-Gil
R W
RW
Department of Medicine, University of California, San Diego 92093.
Olee
T
T
Klink
B K
BK
Kenny
T P
TP
Robbins
D L
DL
Carson
D A
DA
Chen
P P
PP
D000595:Amino Acid Sequence; D001172:Arthritis, Rheumatoid / Q000276:immunology*; D001323:Autoantibodies / Q000235:genetics*; D001483:Base Sequence; D015342:DNA Probes; D015326:Gene Rearrangement, B-Lymphocyte, Heavy Chain; D005803:Genes, Immunoglobulin; D006801:Humans; D007143:Immunoglobulin Heavy Chains / Q000235:genetics; D008969:Molecular Sequence Data; D009154:Mutation; D016133:Polymerase Chain Reaction; D012217:Rheumatoid Factor / Q000235:genetics
D001323:Autoantibodies; D015342:DNA Probes; D007143:Immunoglobulin Heavy Chains; D012217:Rheumatoid Factor