Human leukocyte antigen-B (-Bw6/-Bw4 I 80, T 80) and human leukocyte antigen-C (-C1/-C2) subgrouping using pyrosequence analysis

Elisabetta Ugolotti, Irene Vanni, Alessandro Raso, Fabio Benzi, Mauro Malnati, Roberto Biassoni

Research output: Contribution to journalArticlepeer-review

Abstract

Specific combinations of killer immunoglobulin-like receptors (KIR) and human leukocyte antigen (HLA) class I molecules characterized by a particular residue 80 are significantly associated with outcomes in different pathologic conditions, such as autoimmunity, pathogenic infection, cancer, and reproductive failure. Thus, a simplified method for HLA typing used in association with the analysis of KIR genotype (Kirotype) is of particular interest to extend the analysis of larger series. Here, we describe a quick and inexpensive method that allows use of pyrosequencing, a helpful subtyping of HLA class I molecules, into HLA-Bw6, -Bw4 I 80 or -Bw4 T 80, HLA-C1, or -C2 groups and HLA-A allotypes sharing Bw4+ epitope or the rare HLA-B allotypes displaying the C1 motif. In particular, this analysis is focused on the amino acids around residue 80, known to be relevant in defining the affinity of KIR/HLA interaction and in the functional effects. This method was demonstrated to have good sensitivity, specificity, and reproducibility of detection and it was validated using a panel of HLA-typed International Histocompatibility Workshop (IHW) cell lines and clinical isolates. Using an allele quantitative acquisition mode, the method permitted us to obtain an accurate sequencing as required in heterozygous and/or homozygous sample definition.

Original languageEnglish
Pages (from-to)859-868
Number of pages10
JournalHuman Immunology
Volume72
Issue number10
DOIs
Publication statusPublished - Oct 2011

Keywords

  • Genotyping
  • HLA
  • KIR
  • NK cells
  • Pyrosequencing

ASJC Scopus subject areas

  • Immunology
  • Immunology and Allergy

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