A novel method for KIR-ligand typing by pyrosequencing to predict NK cell alloreactivity

Gong Yun, Jakub Tolar, Anton K. Yerich, Steven G E Marsh, James Robinson, Harriet Noreen, Bruce R. Blazar, Jeffrey S. Miller

Research output: Contribution to journalArticlepeer-review

10 Scopus citations

Abstract

Studies have shown that KIR-ligand mismatching to predict NK cell alloreactivity may result in less relapse and better survival in patients with AML. KIR-ligands are distinguished by single nucleotide polymorphisms (SNPs) from HLA-B and HLA-C sequences. We hypothesized that pyrosequencing to determine KIR-ligand status by direct sequencing of the ligand epitope can be done as an alternative to high-resolution HLA-typing. Pyrosequencing is rapid and would be particularly useful in analysis of retrospective cohorts where high-resolution HLA-typing is unavailable or too expensive. To validate this assay, RNA and DNA from 70 clinical samples were tested for KIR-ligand by pyrosequencing. Primer binding to invariant regions without known SNPs was critical for KIR-ligand assignment by pyrosequencing to be in full concordance with high-resolution HLA-typing. Pyrosequencing is sensitive, specific, high-throughput, inexpensive, and can rapidly screen KIR-ligand status to evaluate potential alloreactive NK cell or transplant donors.

Original languageEnglish (US)
Pages (from-to)272-280
Number of pages9
JournalClinical Immunology
Volume123
Issue number3
DOIs
StatePublished - Jun 2007

Bibliographical note

Funding Information:
This work was supported by National Institutes of Health Grant P01-CA-65493 (JT, BRB, JSM), P01-CA-111412 (JSM, SGEM), R01-HL-55417 (JSM), R01-CA-72669 (BRB) and supported in part by Grant M01-RR00400 from the National Center for Research Resources and the Cancer Center Translational Cell Therapy Core (P30-CA-77598).

Copyright:
Copyright 2009 Elsevier B.V., All rights reserved.

Keywords

  • Alloreactivity
  • HLA-typing
  • Human
  • Killer immunoglobulin receptor
  • NK cells
  • Pyrosequencing

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