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Genetically linked C-type lectin-related ligands for the NKRP1 family of natural killer cell receptors

Nature Immunology volume 4, pages 801–807 (2003)Cite this article

Abstract

The natural killer (NK) gene complex (NKC) encodes orphan lectin-like NK cell receptors that may explain uncharacterized NK cell specificities. Unlike other NKC-encoded receptors that recognize molecules with major histocompatibility complex (MHC) class I folds, here we show that mouse Nkrp1d and Nkrp1f bind specific C-type lectin-related (Clr) molecules. Nkrp1d mediated inhibition when recognizing Clrb, a molecule expressed in dendritic cells and macrophages. Nkrp1 (official gene name, Klrb1) and Clr are intertwined in a genetically conserved NKC region showing recombination suppression, reminiscent of plant self-incompatibility loci. Thus, these findings broaden the 'missing-self' hypothesis from solely involving MHC class I to including related NK cell receptors for lectin-like ligands, and reflect genetic strategies for biological self-recognition processes in other species.

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Figure 1: Reaction of Nkrp1f reporter cells and tetramers with tumor cell lines.
Figure 2: Evolutionary, genomic and functional relationships between Nkrp1 and Clr gene families.
Figure 3: Clrb and Clrg are ligands for Nkrp1d and Nkrp1f, respectively.
Figure 4: Expression and inhibitory function of Nkrp1d in primary NK cells.
Figure 5: Clr expression in primary cells.

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Acknowledgements

We thank J. Laurent, E. Blattenberger and L. Yang for technical support, J. Nasrallah (Cornell University) for discussions, and E. Unanue, M. Colonna, T. Hansen and A. French for comments on the manuscript. Work in the Yokoyama laboratory is supported by the Howard Hughes Medical Institute, the Barnes-Jewish Hospital Foundation and grants from the National Institutes of Health. The Burroughs-Wellcome Fund, Kilo Foundation and National Institutes of Health support work in the Fremont laboratory. O.V.N. is supported by a Cancer Research Institute postdoctoral fellowship.

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Authors and Affiliations

  1. Rheumatology Division, Department of Medicine, Howard Hughes Medical Institute, St. Louis, 63110, Missouri, USA

    Koho Iizuka, Beatrice F M Plougastel & Wayne M Yokoyama

  2. Departments of Pathology and Immunology, Washington University School of Medicine, St. Louis, 63110, Missouri, USA

    Olga V Naidenko, Daved H Fremont & Wayne M Yokoyama

  3. Barnes-Jewish Hospital, St. Louis, 63110, Missouri, USA

    Wayne M Yokoyama

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  1. Koho Iizuka
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  2. Olga V Naidenko
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Correspondence to Wayne M Yokoyama.

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Supplementary information

Supplementary Fig. 1.

Minimal allelic polymorphism of Clr and Nkrp1 between C57BL/6 and 129 mouse strains. (a) Identity of Clr sequences from C57BL/6 (B6) and 129 mice. Amino acid alignments of the B6 (Clrb: NP_444339; Clrg: NP_064653) and 129 alleles from Genbank and Celera databases, respectively, were performed using the BLAST algorithm. The contact residues (specificity determinants) as ascertained by the crystal structure of the Nkrp1f-Clrg complex are indicated by the asterisks. The “V” indicates the start of the Clrg molecule used in the crystallization studies. Note that the Clrb and Clrg sequences are identical. (b) Minimal polymorphism of Nkrp1 molecules from B6 and 129 mice. Amino acid alignment of Nkrp1d and Nkrp1f from B6 and 129 strain mice. Amino acid alignments of the B6 (Nkrp1d, GenBank accession number NP_0750599; Nkrp1f, Genbank NP_694734) and 129 alleles from Genbank and Celera databases, respectively, were performed using the BLAST algorithm. The contact residues (specificity determinants) as ascertained by the crystal structure of the Nkrp1f-Clrg complex are indicated by the asterisks. The “V” indicates the start of the Nkrp1f molecule used in the crystallization studies. Note that the Nkrp1f sequences are identical and that none of the predicted contact sites for Nkrp1d is a polymorphic residue. (PDF 40 kb)

Supplementary Fig. 2.

Specificity of mAb 2D12 for Nkrp1d. Reporter cells for Nkrp1a, Nkrp1c, Nkrp1d or Nkrp1f were produced by retroviral transduction of chimeric molecules containing Nkrp1 ectodomains, Ly49a transmembrane, and CD3z cytoplasmic domains, as indicated. The retroviral expression vectors contained GFP in the second cistron. GFP-positive cells were sorted to uniformity, then were mixed with untransduced BWZ.36 parental cells for staining with 2D12 mAb followed by PE-conjugated goat anti-mouse IgG antibody. Cells were then analyzed with FACS. (PDF 42 kb)

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Iizuka, K., Naidenko, O., Plougastel, B. et al. Genetically linked C-type lectin-related ligands for the NKRP1 family of natural killer cell receptors. Nat Immunol 4, 801–807 (2003). https://doi.org/10.1038/ni954

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