Efficient isolation of cDNA clones encoding rheumatoid arthritis autoantigens by lambda phage surface display.
Bacteriophage lambda surface display was used to isolate cDNA clones encoding autoantigens recognized by synovial fluid (SF) or sera from patients with rheumatoid arthritis (RA). We constructed cDNA libraries from human synovial sarcoma cells and synovial tissue, using the surface display vector lambdafoo. The cDNA libraries were screened by affinity selection using 40 SF and 44 sera as probes separately immobilized in microtiter wells. Phage clones isolated encode 13 different autoantigens; an unknown protein, two proteins previously unanalyzed as autoimmune antigens, three proteins previously unknown to be recognized by RA sera, and seven known RA antigens. When analyzed their sensitivity and specificity for RA by phage enzyme-linked immunosorbent assay, frequencies of sera that recognize the newly-isolated autoantigens ranged from 20.5 to 6.8% of a panel of RA sera, and 13.6-0% of other autoimmune disease sera. These results indicate that the lambda phage surface display may be powerful for the isolation of cDNA clones encoding autoantigens recognized by SF or sera from patients with not only RA but also other autoimmune diseases.
Tsukuba Research Laboratory, Toagosei Co. Ltd., 2 Ohkubo, Tsukuba, Ibaraki 300-2611, Japan.
Mikio
M
Niwa
Kazuhito
K
Fukuoka
Takashi
T
Fujimoto
Ichiro N
IN
Maruyama
D001172:Arthritis, Rheumatoid / Q000276:immunology*; D001324:Autoantigens / Q000235:genetics* / Q000276:immunology*; D010582:Bacteriophage lambda / Q000235:genetics*; D002460:Cell Line; D003001:Cloning, Molecular / Q000379:methods; D018076:DNA, Complementary / Q000235:genetics* / Q000302:isolation & purification*; D015723:Gene Library; D006801:Humans; D019151:Peptide Library; D013584:Sarcoma, Synovial / Q000235:genetics
D001324:Autoantigens; D018076:DNA, Complementary; D019151:Peptide Library