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14i NKT Cells1
* Ludwig Institute for Cancer Research, Epalinges, Switzerland; and
Experimental Immunology Unit, Cancer Immunotherapy and Gene Therapy Program, Department of Oncology, Dipartimento di Biotecnologie, H. San Raffaele Scientific Institute, Milan, Italy
CD1d-dependent invariant V
14 (V14i) NKT cells are innate T lymphocytes expressing a conserved semi-invariant TCR, consisting, in mice, of the invariant V14-J18 TCR -chain paired mostly with V
8.2 and V7. The cellular requirements for thymic positive and negative selection of V14i NKT cells are only partially understood. Therefore, we generated transgenic mice expressing human CD1d (hCD1d) either on thymocytes, mainly CD4+ CD8+ double positive, or on APCs, the cells implicated in the selection of V14i NKT cells. In the absence of the endogenous mouse CD1d (mCD1d), the expression of hCD1d on thymocytes, but not on APCs, was sufficient to select V14i NKT cells that proved functional when activated ex vivo with the Ag -galactosyl ceramide. V14i NKT cells selected by hCD1d on thymocytes, however, attained lower numbers than in control mice and expressed essentially V8.2. The low number of V8.2+ V14i NKT cells selected by hCD1d on thymocytes was not reversed by the concomitant expression of mCD1d, which, instead, restored the development of V7+ V14i NKT cells. V8.2+, but not V7+, NKT cell development was impaired in mice expressing both hCD1d on APCs and mCD1d. Taken together, our data reveal that selective CD1d expression by thymocytes is sufficient for positive selection of functional V14i NKT cells and that both thymocytes and APCs may independently mediate negative selection.
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