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

doi:10.1016/j.clim.2007.01.011

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 journal › Article › peer-review

11 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 language	English (US)
Pages (from-to)	272-280
Number of pages	9
Journal	Clinical Immunology
Volume	123
Issue number	3
DOIs	https://doi.org/10.1016/j.clim.2007.01.011
State	Published - 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).

Keywords

Alloreactivity
HLA-typing
Human
Killer immunoglobulin receptor
NK cells
Pyrosequencing

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10.1016/j.clim.2007.01.011

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title = "A novel method for KIR-ligand typing by pyrosequencing to predict NK cell alloreactivity",

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.",

keywords = "Alloreactivity, HLA-typing, Human, Killer immunoglobulin receptor, NK cells, Pyrosequencing",

author = "Gong Yun and Jakub Tolar and Yerich, {Anton K.} and Marsh, {Steven G E} and James Robinson and Harriet Noreen and Blazar, {Bruce R.} and Miller, {Jeffrey S.}",

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).",

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doi = "10.1016/j.clim.2007.01.011",

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AU - Yun, Gong

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AU - Marsh, Steven G E

AU - Robinson, James

AU - Noreen, Harriet

AU - Blazar, Bruce R.

AU - Miller, Jeffrey S.

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PY - 2007/6

Y1 - 2007/6

N2 - 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.

AB - 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.

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KW - NK cells

KW - Pyrosequencing

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

Abstract

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Keywords

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