title: Allelic Polymorphism Determines Surface Expression or Intracellular Retention of the Human NK Cell Receptor KIR2DL5A (CD158f).
abstract: KIR2DL5 (CD158f) is the most recently identified inhibitory member of human killer-cell Ig-like receptors (KIRs), which enable NK cells to sense self-HLA. Unlike KIR2DL1-3, recognizing HLA-C allotypes through Ig-like domains of the D1-D2 type, KIR2DL5 shares a D0-D2 configuration with KIR2DL4, and its ligands have not been identified. KIR2DL5 is encoded by two paralogous genes displaying copy number variation and allelic polymorphism-KIR2DL5AandKIR2DL5B. UP-R1 mAb, raised against the common alleleKIR2DL5A*001, enables specific KIR2DL5 detection. However, not everyKIR2DL5+individual has NK cells staining with UP-R1, discrepancy explained in part by epigenetically silentKIR2DL5Balleles with a distinctive substitution in a promoter RUNX-binding site. Furthermore, we show here that the transcribed alleleKIR2DL5A*005, second most common of its locus, fails to confer NK cells UP-R1 reactivity, phenotype explained by inefficacious transport of its product to the cell surface. Two amino acid substitutions distinguish theKIR2DL5A*005and*001coding regions. Western blot, flow cytometry, and confocal microscopy analyses of cells transfected with tagged constructs demonstrate that a serine substitution for glycine-174, conserved in most KIR, is mainly responsible for KIR2DL5A*005 intracellular retention, and it also affects mAb recognition. In contrast, substitution of aspartate for asparagine 152 has only a minor effect on surface expression, despite destroying an otherwise conserved N-glycosylation site. Our results help to explain the variable expression profile ofKIR2DL5+subjects and indicate that functional polymorphisms in both its promoter and its coding regions are critical for understanding the KIR2DL5 role in immunity and its importance for human health.
authors.affiliation: Immunogenetics and Histocompatibility, Instituto de Investigación Sanitaria Puerta de Hierro , Madrid , Spain.
authors.forename: Elisa
authors.initials: E
authors.lastname: Cisneros
authors.affiliation: Immunogenetics and Histocompatibility, Instituto de Investigación Sanitaria Puerta de Hierro , Madrid , Spain.
authors.forename: Ernesto
authors.initials: E
authors.lastname: Estefanía
authors.affiliation: Immunogenetics and Histocompatibility, Instituto de Investigación Sanitaria Puerta de Hierro , Madrid , Spain.
authors.forename: Carlos
authors.initials: C
authors.lastname: Vilches
keywords: KIR; NK-cell receptors; allelic polymorphism; natural killer cells; repertoire development