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Molecular and Genetic Basis for Strain-Dependent NK1.1 Alloreactivity of Mouse NK Cells¹

James R. Carlyle^2,*, Aruz Mesci^*, Belma Ljutic^3,*, Simon Belanger, Lee-Hwa Tai, Etienne Rousselle, Angela D. Troke, Marie-France Proteau and Andrew P. Makrigiannis^2,

^* Department of Immunology, University of Toronto, Sunnybrook and Women’s Research Institute, Toronto, ON, Canada; Laboratory of Molecular Immunology, Institut de Recherches Cliniques de Montréal, Montréal, QC, Canada

NK1.1 alloantigen expression can be used to define NK cells in certain mouse strains, such as B6 (NKR-P1C) and SJL (NKR-P1B). However, BALB/c NK cells do not react with the anti-NK1.1 mAb, PK136. To investigate the NK1.1^– phenotype of BALB/c NK cells, we have undertaken NK1.1 epitope mapping and genomic analysis of the BALB/c Nkrp1 region. Bacterial artificial chromosome library analysis reveals that, unlike the Ly49 region, the Nkrp1-Ocil/Clr region displays limited genetic divergence between B6 and BALB/c mice. In fact, significant divergence is confined to the Nkrp1b and Nkrp1c genes. Strikingly, the B6 Nkrp1d gene appears to represent a divergent allele of the Nkrp1b gene in BALB/c mice and other strains. Importantly, BALB/c NK cells express abundant and functional Nkrp1 transcripts, and the BALB/c NKR-P1B receptor functionally binds Ocil/Clr-b ligand. However, the BALB/c NKR-P1B/C sequences differ from those of the known NK1.1 alloantigens, and epitope mapping demonstrates that directed mutation of a single amino acid in the NKR-P1B^BALB protein confers NK1.1 reactivity. Thus, PK136 mAb recognizes, in part, a distal C-terminal epitope present in NKR-P1B^Sw/SJL and NKR-P1C^B6, but absent in NKR-P1A/D/F^B6 and NKR-P1B/C^BALB. Allelic divergence of the Nkrp1b/c gene products and limited divergence of the BALB/c Nkrp1-Ocil/Clr region explain a longstanding confusion regarding the strain-specific NK1.1 alloantigen reactivity of mouse NK cells.

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