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R. Mathew, C. Watte, C. Kan, J. Stein-Streilein; Surrogate Acaid Antigen Presenting Cells Induce Ly49c/i Dependent Il-10 Production in INkt Cells. Invest. Ophthalmol. Vis. Sci. 2008;49(13):2520.
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NKT cells express various NK cell-associated molecules including proteins in the Ly49 receptor family. Ly49 C/I is an inhibitory receptor that when ligated inhibits cytolytic activity and cytokine production by NK and NKT cells Recently, in vivo studies in our laboratory established that Ly49 C/I invariant (i)NKT cells were required for induction of peripheral tolerance via the eye (Anterior chamber associated immune deviation, ACAID).The purpose of this study was to analyze the role of Ly49 C/I on iNKT cells in the production of immunosuppressive cytokines (IL-10) post exposure to tolerogenic APC( surrogate ACAID APC).
Antigen presenting cells (APC) were derived from either peritoneal exudate cells (PEC) or bone marrow cell cultures. Incubation of the APC with OVA or TGF β and OVA generated immunogenic or tolerogenic APC, respectively The resulting APC were then incubated with iNKT cells ( 24.8A hybridoma) for varying time periods with or without anti-LY49 (5E6). FACS analysis, ELISA and semi-quantitative RT-PCR were used to measure protein expression and mRNA expression of cytokines (IL-10 and IFNγ) relative to the house keeping genes.
iNKT cells (24.8A hybridoma) showed an increase in IL-10 mRNA expression and decrease in IFNγ after one hour incubation with tolerogenic APC. iNKT cells responded to immunogenic APC with increased IFNγ mRNA and little or no IL-10 mRNA levels.
These results support the postulate that ligation of Ly49 C/I on iNKT cells or their hybridoma counterparts not only inhibits inflammatory cytokine ( IFNγ) production, but at the same time contributes to the production of immunosuppressive cytokines, such as IL-10. Thus the role of Ly49 C/I is enlarged and includes this novel role in the induction of peripheral tolerance. The research was supported in part by NIH grant : EY0111983
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