Elsevier

Human Immunology

Volume 64, Issue 5, May 2003, Pages 567-571
Human Immunology

A Multi-Laboratory characterization of the KIR genotypes of 10th International Histocompatibility Workshop cell lines

https://doi.org/10.1016/S0198-8859(03)00042-9Get rights and content

Abstract

Killer immunoglobulinlike receptors (KIRs) are expressed on natural killer and T cells. Both inhibitory and noninhibitory forms have been described, leading to inhibition or continuation of cellular killing activity. The natural ligands identified so far of KIRs are class I human leukocyte antigens (HLA). In particular, the interaction of some KIRs with HLA-Cw has been well characterized. Recent work has implicated KIRs in affecting the outcome of hematopoietic stem-cell transplant (HSCT). This may well lead to a requirement for prospective KIR typing of donor and recipient. We have utilized different typing systems (two using polymerase chain reaction–sequence-specific primers, and one using polymerase chain reaction–sequence-specific oligonucleotide probes) in three separate laboratories to characterize the KIR gene complement of 25 cell lines from the 10th International Histocompatibility Workshop. There were consistent results in 22, and minor differences in 3. When compared with previous results for some of these cell lines, no further differences were found. The differences are due to typing of KIRs KIR2DL1 and KIR2DS5, and may be explained by technical differences or the inability to type new variants. Further improvements in typing may be required if population and clinical studies are to produce accurate results.

Section snippets

Abbreviations

    GVHD

    graft-versus-host disease

    HLA

    human leukocyte antigen

    HSCT

    hematopoietic stem-cell transplant

    IHW

    International Histocompatibility Workshop

    KIR

    killer immunoglobulin-like receptor

    PCR-SSP

    polymerase chain reaction–sequence-specific primers

    PCR-SSOP

    polymerase chain reaction–sequence-specific oligonucleotide probes

Samples

DNA was extracted from 25 cell lines from the 10th IHW (illustrated in Figure 1) and distributed to all three laboratories. The results for all three laboratories were collated in Laboratory 3.

Laboratory 1 (Guy’s Hospital)

The PCR-SSP method as described by Uhrberg et al. [6] was used, with additional primers for 2DL4, 2DL5, 2DS3, 2DS5, and 2DL1v as previously described 8, 9.

Laboratory 2 (Belfast)

The PCR-SSOP method as previously described was used [2].

Laboratory 3 (Birmingham)

The PCR-SSP method using modified primers as described by Uhrberg et al. [6] was used. In

Results

The results for the KIR genotypes are given in Figure 1. Using the nomenclature originated by Uhrberg et al. [6] then Witt et al. [13] and expanded by Norman et al. 8, 9, the most common KIR profile is that originally designated AA1. There is complete agreement between the three laboratories for 22 of the cell lines tested. There are minor differences between the three laboratories for the following three of cell lines: BOLETH (10th IHW number 9031), HOR (9053), and LBUF (9048). The genes of

Discussion

We have genotyped 25 cell lines from the 10th IHW and obtained consistent results by different methods in three laboratories for 22 of them, with minor differences between the remaining three. We have chosen these cell lines because we believe them to be widely available among the HLA typing community. There was some overlap between the cell lines we tested and those tested by Gómez-Lozano [7], but no further differences in KIR profile were introduced when all four sets of results were compared.

Acknowledgements

Many thanks to Alasdair Heads and Robert Collins for cell culture and DNA extraction.

References (15)

There are more references available in the full text version of this article.

Cited by (0)

View full text
Copyright © 2003 American Society for Histocompatibility and Immunogenetics. Published by Elsevier Inc. All rights reserved.