Yoshiho Nagata

HLA-DP typing by analysis of DNA restriction fragment length polymorphisms in the HLA-DPβ subregion

HLA class II antigens are encoded in the HLA-D region and are highly polymorphic. Southern hybridization technique was used to analyze restriction fragment length polymorphisms (RFLPs) in the DP beta gene and an attempt was made to correlate these with DP haplotypes derived from primed lymphocyte typing (PLT) analysis. Digestion of DNA from 32 Epstein-Barr virus (EBV)-transformed cell lines (of haplotypes DPw2, DPw3, DPw4, DPw5, and Cp63) with three different restriction endonucleases. Southern transfer, and hybridization to the DP beta cDNA probe revealed multiple fragments in all cell lines tested. The polymorphic patterns of these fragments were found to correlate with DP haplotypes, suggesting the possibility that the analysis of DNA RFLPs (DNA typing) in the HLA-DP beta subregion can distinguish and identify HLA-DP haplotypes.

No difference in the nucleotide sequence of the DQß ß1 domain between narcoleptic and healthy individuals with DR2,Dw2

Abstract Narcolepsy is a sleep disorder completely associated with HLA-DR2,Dw2. We demonstrated the 100% presence of three DQB fragments (EcoRI 2.4-kb, BamHI 2.9-kb, and PstI 12-kb) in narcoleptic patients that were detected in healthy DR2 controls only at decreased frequencies (1). In this paper, we have cloned the DQB gene from three Japanese narcoleptic patients and sequenced their s1 domain in order to study the sequence polymorphisms that might exist in the DQs genes of patients, but no difference in sequence could be found between narcoleptic and healthy individuals, suggesting that narcolepsy is not due to mutation in the DQs gene. In this context, a possible role of the HLA class II antigens in narcolepsy is discussed.