Generation of anti-HLA-DR4 specific antibodies by immunization of the recombinantly expressed allelic subtype-specific region of the HLA-DRB1 0405 molecules

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Title
Generation of anti-HLA-DR4 specific antibodies by immunization of the recombinantly expressed allelic subtype-specific region of the HLA-DRB1 0405 molecules
Author(s)
Jung Hyun Park; Eun Wie Cho; Yun Jung Lee; Jin Chung; Kyung Soo Hahm; Kil Lyong Kim
Bibliographic Citation
BMB Reports, vol. 31, no. 2, pp. 111-116
Publication Year
1998
Abstract
HLA-DR4 is the dominant allele of MHC class II genes in Koreans. In particular, the DRB1*0405 subtype has been reported to be almost exclusively expressed in Far East Asians, and has also been observed to be strongly associated with rheumatoid arthritis in Koreans and the Japanese. Identification of this specific allele has been mainly performed by PCR-based methods, which is often time consuming, costly, and involves tedious procedures such as the isolation of genomic DNA, PCR, and gel electrophoresis. To develop a more convenient tool for screening vast amounts of samples as well as to generate reagents which might also be used in other applications, in this study, antibodies were produced against this specific HLA subtype. By PCR, an allele-specific region covering the β1 domain of DRB1*0405 was amplified and recombinantly expressed in E.coli. Immunization of Lewis rats with the purified protein yielded an allele specific antiserum. Western blot analysis showed the selective detection of the HLA-DR β-chain. Using this antiserum, established cell lines and peripheral blood lymphocytes were analyzed on their HLA haplotype by fluorescence activated flow cytometry. These novel antibodies will provide a powerful tool in the detection and investigation of DR4 alleles.
Keyword
Allelic typingAntibodiesHLA-DRMHC Class IIRecombinant protein
ISSN
1225-8687
Publisher
Korea Soc-Assoc-Inst
Type
Article
Appears in Collections:
Division of Biomedical Research > Rare Disease Research Center > 1. Journal Articles
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