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 journalArticle

10 Citations (Scopus)

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 1 2007

Fingerprint

Natural Killer Cells
Ligands
Histocompatibility Testing
Single Nucleotide Polymorphism
HLA-C Antigens
HLA-B Antigens
Epitopes
Cohort Studies
Tissue Donors
RNA
Transplants
Recurrence
Survival
DNA

Keywords

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

Cite this

A novel method for KIR-ligand typing by pyrosequencing to predict NK cell alloreactivity. / Yun, Gong; Tolar, Jakub; Yerich, Anton K.; Marsh, Steven G E; Robinson, James; Noreen, Harriet; Blazar, Bruce R.; Miller, Jeffrey S.

In: Clinical Immunology, Vol. 123, No. 3, 01.06.2007, p. 272-280.

Research output: Contribution to journalArticle

Yun, Gong ; Tolar, Jakub ; Yerich, Anton K. ; Marsh, Steven G E ; Robinson, James ; Noreen, Harriet ; Blazar, Bruce R. ; Miller, Jeffrey S. / A novel method for KIR-ligand typing by pyrosequencing to predict NK cell alloreactivity. In: Clinical Immunology. 2007 ; Vol. 123, No. 3. pp. 272-280.
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