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The Journal of Immunology, 2001, 167: 4180-4186.
Copyright © 2001 by The American Association of Immunologists

Regulation of NKT Cells by Ly49: Analysis of Primary NKT Cells and Generation of NKT Cell Line1

Motoi Maeda*, Stefan Lohwasser*, Takashi Yamamura{dagger} and Fumio Takei2,*,{ddagger}

* Terry Fox Laboratory, British Columbia Cancer Agency, Vancouver, British Columbia, Canada; {dagger} Department of Immunology, National Institute of Neuroscience, Kodaira, Tokyo, Japan; and {ddagger} Department of Pathology and Laboratory Medicine, University of British Columbia, Vancouver, British Columbia, Canada

TCR{alpha}{beta}+NK1.1+ (NKT) cells are known to express various NK cell-associated molecules including the Ly49 family of receptors for MHC class I, but its functional significance has been unclear. Here, we examined the expression of Ly49A, C/I and G2 on various NKT cell populations from normal and MHC class I-deficient C57BL/6 mice as well as their responsiveness to {alpha}-galactosylceramide ({alpha}-GalCer), a potent stimulator of CD1d-restricted NKT cells. The frequency and the level of Ly49 expression varied among NKT cells from different tissues, and were regulated by the expression of MHC class I and CD1d in the host. Stimulation of various NKT cells with {alpha}-GalCer suggested that Ly49 expression inversely correlates with the responsiveness of NKT cells to {alpha}-GalCer. Moreover, {alpha}-GalCer presented by normal dendritic cells stimulated purified Ly49-, but not Ly49+, splenic NKT cells, whereas MHC class I-deficient dendritic cells presented {alpha}-GalCer to both Ly49+ and Ly49- NKT cells equally well. Therefore, MHC class I on APCs seems to inhibit activation of NKT cells expressing Ly49. To further characterize CD1d-restricted NKT cells, we generated an {alpha}-GalCer-responsive NKT cell line from thymocytes. The line could only be generated from Ly49-NK1.1+CD4+ thymocytes but not from other NKT cell subsets, and it lost expression of NK1.1 and CD4 during culture. Together, these results indicate the functional significance of Ly49 expression on NKT cells.




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