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Dramatically reduced surface expression of NK cell receptor KIR2DS3 is attributed to multiple residues throughout the molecule

Genes & Immunity volume 10, pages 162–173 (2009)Cite this article

Abstract

Using flow cytometry, fluorescent microscopy and examination of receptor glycosylation status, we demonstrate that an entire killer cell immunoglobulin-like receptor (KIR) locus (KIR2DS3)—assumed earlier to be surface expressed—appears to have little appreciable surface expression in transfected cells. This phenotype was noted for receptors encoded by three allelic variants including the common KIR2DS3*001 allele. Comparing the surface expression of KIR2DS3 with that of the better-studied KIR2DS1 molecule in two different cell lines, mutational analysis identified multiple polymorphic amino-acid residues that significantly alter the proportion of molecules present on the cell surface. A simultaneous substitution of five residues localized to the leader peptide (residues −18 and −7), second domain (residues 123 and 150) and transmembrane region (residue 234) was required to restore KIR2DS3 to the expression level of KIR2DS1. Corresponding simultaneous substitutions of KIR2DS1 to the KIR2DS3 residues resulted in a dramatically decreased surface expression. Molecular modeling was used to predict how these substitutions contribute to this phenotype. Alterations in receptor surface expression are likely to affect the balance of immune cell signaling impacting the characteristics of the response to pathogens or malignancy.

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Acknowledgements

Funding from the Office of Naval Research N00014-05-1-0784 and N00014-06-1-0726 supported this research. The views expressed in this paper are those of the authors and do not reflect the official policy of the Department of Navy, the Department of Defense or the United States government. These studies were conducted using the Tissue Culture, Flow Cytometry and Cell Sorting, Macromolecular Analysis, and Microscopy and Imaging Shared Resources of Lombardi Comprehensive Cancer Center. Computing time and support were in part provided by the Advanced Biomedical Computing Center at the National Cancer Institute (Frederick, MD, USA). Molecular graphics images were produced using the UCSF Chimera package from the Resource for Biocomputing, Visualization, and Informatics at the University of California, San Francisco. We thank Thanemozhi Govindan, Michelle Makiya and Karen Creswell for providing excellent technical assistance.

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Authors and Affiliations

  1. Department of Oncology, Lombardi Cancer Center, Georgetown University Medical Center, Washington, DC, USA

    C J VandenBussche, T J Mulrooney, W R Frazier, S Dakshanamurthy & C K Hurley

  2. Tumor Biology Training Program, Lombardi Cancer Center, Georgetown University Medical Center, Washington, DC, USA

    C J VandenBussche, T J Mulrooney & W R Frazier

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  1. C J VandenBussche
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  2. T J Mulrooney
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  4. S Dakshanamurthy
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  5. C K Hurley
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Correspondence to C K Hurley.

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VandenBussche, C., Mulrooney, T., Frazier, W. et al. Dramatically reduced surface expression of NK cell receptor KIR2DS3 is attributed to multiple residues throughout the molecule. Genes Immun 10, 162–173 (2009). https://doi.org/10.1038/gene.2008.91

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