Comprehensive HLA‐DP typing using polymerase chain reaction with sequence‐specific primers and 95 sequence‐specific primer mixes

Abstract

HLA-DP is the third of the class II molecules. Its role is antigen presentation, and it has been suggested to play a part in the susceptibility to certain diseases such as berylliosis, sarcoidosis and juvenile chronic arthritis. The standard typing method is SSO typing, although other methods have been used. Probably the best is sequence-based typing, but this is time-consuming and requires expensive equipment. We describe a method for comprehensive HLA-DPB1 and HLA-DPA1 typing using sequence-specific primers. This method has the advantages that it is rapid - typing a single DNA sample takes under 3 hours - and does not require any special equipment or reagents. The method has been shown to be highly accurate by typing 60 cell line DNA samples in which there was 100% agreement between the types obtained and the published information. Similarly typing of 20 DNA samples previously typed by sequence-based typing gave 100% concordance. We used the method to type DNA samples from 102 UK Caucasoid kidney donors. The allele frequencies agree with previously published data. Linkage disequilibria between HLA-DPB1, HLA-DPA1 and the other class II antigens have been investigated. Strong linkage disequilibria exist between certain HLA-DPB1 and HLA-DPA1 alleles. This is unsurprising in view of their proximity on the chromosome. More unexpectedly, the data also suggest that genes further away along the chromosome are in linkage disequilibrium with HLA-DP, forming extended haplotypes.

Analysis of MHC Encoded Antigen-processing Genes TAP1 and TAP2 Polymorphisms in Sarcoidosis

“…This allowed associations between different alleles to be analyzed-a strong association giving good evidence that these alleles form a haplotype (16).…”

“…To characterize the HLA-DPB1 alleles present in the UK control and patient samples a Sequence Specific Primer (SSP-PCR) method was used as described previously (16).…”

Beryllium presentation to CD4+T cells underlies disease-susceptibility HLA-DP alleles in chronic beryllium disease

“…Our data suggest that the polymorphism at positions 55-56 may be more important for presentation and possibly susceptibility because every presenting allele showed this difference. Importantly, among frequently inherited HLA-DP alleles, the glutamate at position 69 and the negatively charged residues at positions 55-56 frequently coexist (31), and both may contribute to disease susceptibility. Position 57 in DQ␤ (corresponding to position 55 in DP␤; ref.…”

Beryllium Presentation to CD4+ T Cells Is Dependent on a Single Amino Acid Residue of the MHC Class II β-Chain

“…In addition to exposure, genetic susceptibility plays a major role in disease development, with Glu 69 -containing HLA-DP alleles being strongly associated with the development of beryllium sensitization (13)(14)(15)25). However, in the most commonly inherited HLA-DP alleles, the Glu 69 and the negatively charged residues at positions 55-56 frequently coexist (26), raising the possibility that both polymorphisms contribute to disease susceptibility. Our previous work also suggested the possibility that the 55-56 polymorphism could present beryllium to T cells and stimulate T cell proliferation (19).…”