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Efficient isolation of cDNA clones encoding rheumatoid arthritis autoantigens by lambda phage surface display
Mikio Niwa1, Kazuhito Fukuoka, Takashi Fujimoto
1Tsukuba Research Laboratory, Toagosei Co. Ltd., 2 Ohkubo, Tsukuba, Ibaraki 300-2611, Japan.
Journal of Biotechnology
|October 7, 2004
Summary
Bacteriophage lambda display identified 13 autoantigens in rheumatoid arthritis (RA) patients. This method aids in discovering novel autoantigens for RA and other autoimmune diseases.
Area of Science:
- Immunology
- Molecular Biology
- Biotechnology
Background:
- Rheumatoid arthritis (RA) is an autoimmune disease characterized by the presence of autoantibodies.
- Identifying specific autoantigens is crucial for understanding RA pathogenesis and developing diagnostic tools.
Purpose of the Study:
- To isolate cDNA clones encoding autoantigens recognized by synovial fluid (SF) and sera from RA patients.
- To evaluate the potential of lambda phage surface display for autoantigen discovery in autoimmune diseases.
Main Methods:
- Construction of cDNA libraries from human synovial sarcoma cells and synovial tissue using the lambda phage surface display vector lambdafoo.
- Screening of cDNA libraries by affinity selection using SF and sera from RA patients as probes.
- Analysis of isolated phage clones for autoantigen identification and characterization.
Main Results:
- Isolation of 13 distinct autoantigens, including novel and known RA autoantigens.
- Newly identified autoantigens showed varying frequencies of recognition by RA sera (6.8-20.5%) and minimal recognition by sera from other autoimmune diseases (0-13.6%).
Conclusions:
- Lambda phage surface display is a powerful technique for isolating autoantigens in RA.
- This method holds promise for the discovery of autoantigens in other autoimmune conditions.