title: Aberrant expression of HLA-B*3565Q is associated with a disrupted disulfide bond.
abstract: The identification of expression variants is a challenge in HLA diagnostics. We here describe the identification of the novel allele HLA-B*3565Q. The serological HLA class I type, as determined by a lymphocytotoxicity test, was A11,24; B38; Bw4; Cw-; whereas PCR-sequence-specific primers resulted in A*11,*24, B*35,*38; Cw*12, thus suggesting the presence of a nonexpressed B*35 allele. To clarify the lack of serological HLA-B35 reactivity, exons 2 and 3 were sequenced following haplotype-specific amplification. At position 564 from the beginning of the coding region (exon 3), a transversion (C-->G) was observed, which, at the amino acid level, results in a substitution from cysteine to tryptophane at position 164 of the mature polypeptide. Because this position is essential for the formation of a disulfide bond linking the cysteine residues at positions 101 and 164, which is strongly conserved in functional class I molecules of vertebrates, the disruption of this bond is very likely to be the reason for the lack of serological detectability. We later found the same novel allele in a second unrelated individual, of whom we were able to establish a lymphoblastoid cell line (B-LCL). Serological testing of this B-LCL indicated a very low aberrant expression of HLA-B*3565Q, which cannot be expected to be detected by standard serology techniques.
authors.affiliation: Labor Dres. Fenner & Partner, Bergstr. 14, 20095 Hamburg, Germany.
authors.forename: Holger-Andreas
authors.initials: HA
authors.lastname: Elsner
authors.forename: Peter A
authors.initials: PA
authors.lastname: Horn
authors.forename: Constanze
authors.initials: C
authors.lastname: Schoenemann
authors.forename: Wolfgang W
authors.initials: WW
authors.lastname: Altermann
authors.forename: Rainer
authors.initials: R
authors.lastname: Blasczyk
mesh_terms: D000483:Alleles; D000595:Amino Acid Sequence; D019943:Amino Acid Substitution; D001483:Base Sequence; D003545:Cysteine / Q000737:chemistry / Q000235:genetics*; D004220:Disulfides / Q000737:chemistry*; D005091:Exons / Q000235:genetics; D015235:HLA-B Antigens / Q000737:chemistry / Q000235:genetics* / Q000276:immunology*; D015797:HLA-B35 Antigen; D006801:Humans; D008969:Molecular Sequence Data; D014364:Tryptophan / Q000737:chemistry / Q000235:genetics
chemical_list: D004220:Disulfides; D015235:HLA-B Antigens; C516140:HLA-B*35:65Q antigen; D015797:HLA-B35 Antigen; D014364:Tryptophan; D003545:Cysteine