An Apparent Functional Correlation Between Variations in Amino Acid Residues in Hla‐dr 4.1 and 4.2 Serological Subtypes and Oligonucleotide Characterization*

Abstract

HLA-DR4 can be subdivided serologically into two specificities, DR4.1 and DR4.2, using well-defined monospecific alloantisera used in the 11th International Histocompatibility Workshop. In this study, a total of 1095 random DR4-positive individuals from several ethnic groups were tested first for serotype DR4.1/4.2 and then for DRB1*04 alleles using polymerase chain reaction (PCR) followed by sequence-specific oligonucleotide probe hybridization (SSOPH). An almost 100% correlation between samples testing positive for DR4.1 and the presence of alanine at position 74 was observed, while samples testing positive for DR4.2 correlated with the presence of glutamic acid at position 74. DRB1*04 alleles 0401, 0402, 0404, 0405, 0408, 0409 and 0410 are aligned in functional groups which coincide with the serological subtype of DR4.1. DRB1*04 alleles 0403, 0406, 0407 and 0411 coincide with subtype DR4.2. Amino acid substitutions at positions 57, 71 and 86 indicate other significant variations between alleles within the serological subgroup of DR4.1 and define five minor subgroups. The serologic and oligonucleotide allelic subgroups are in turn correlated with recognized cellular Dw antigens. While sequence data provide evidence of structural differences, data on cellular antigens support a functional association between these designated groups and their significance in transplantation and GVHD. Testing results are categorized by ethnic group in order to establish frequency data for donor selection criteria.

Confirmation of a recombinant allele B*5603 and a hypothetical reciprocal hybrid

“…Peripheral blood lymphocytes (PBL) were isolated from healthy volunteer donors on a randomized basis. Genomic DNA was extracted from PBLs as described previously (Lee et al. , 1996, 1999).…”

HLA‐DRB1 genotyping in patients with juvenile idiopathic arthritis in Taiwan

“…Genomic DNA was extracted from peripheral blood using the salting‐out method (Miller et al , 1988). Polymerase chain reaction (PCR) and sequence‐specific oligonucleotide probes (SSO) typing methods were used, as in previous studies (Lee et al , 1996; Huang et al , 2001b). In the PCR reaction, primers used for DRB1 amplification were 5′‐GAT CCT TCG TGT CCC CAC AGC ACG‐3′ and 5′‐G CCG CTG CAC TGT GAA GCT CTC‐3′.…”

Human leukocyte antigen phenotypes and hepatitis C viral load

“…To study the relationship between HLA phenotypes and HCV viral load, we have determined the phenotypes of HLA-A, -B, and -DRB1 for all anti-HCV positive subjects by sequence-specific oligonucleotide probe hybridization (SSOPH) (Lee et al, 1996(Lee et al, , 1999Wu et al, 2004). Due to small sample size for subjects with HBsAg-positivity, we subsequently restricted further analyses to HBsAg-negative subjects only.…”

“…HLA phenotypes of all subjects were determined by sequence-specific oligonucleotide probe hybridization (SSOPH) (Lee et al, 1996(Lee et al, , 1999Wu et al, 2004).…”