The rapid iNKT cell response to sensitization is at least partially because of rapidly changing characteristics of stimulatory hepatic lipids. An alternate, but not mutually exclusive, hypothesis is that the expression level of CD1d increases, thereby enabling enhanced iNKT cell activation. Interaction with hepatocytes via CD1d is thought to promote IL-4 secretion by iNKT cells [28]. Using flow cytometry, we examined whether the expression level of CD1d by

hepatocytes in wild-type BALB/c mice changes 1 h after sensitization. (Examination at 30 min was not possible for technical reasons.) A non-significant increase in the hepatocyte CD1d expression level was observed following skin sensitization (Fig. 3). The actual CD1d increase may be even less when considering that small numbers of contaminating APC (Kupffer cells or dendritic cells) may remain attached to the hepatocytes. Furthermore, given that hepatocytes constitute approximately DNA Synthesis inhibitor 10% of CD1d-expressing LMNC, it is difficult

to draw mechanistic conclusions. Still, a role of hepatocytes in iNKT cell activation cannot BMS-907351 mw be excluded entirely. The non-significant increase in CD1d expression by hepatic APCs may contribute to iNKT cell activation. In addition, the levels seen here may represent one point along a down-trending slope that peaked earlier. CD1d appears essential to iNKT cell activation, but whether the critical molecular interaction in our model occurs in vitro (during incubation of iNKT cells with lipids) or in vivo (following adoptive cell transfer) remained unclear. To investigate the importance of endogenous CD1d expression, we compared CS reactions between Jα18−/− and CD1d−/− mice after adoptive transfer of activated wild-type iNKT cells into each group. Both knockout strains are deficient in iNKT cells: Jα18 is a key component of the invariant TCR, while CD1d is essential for the development and activation of iNKT cells [29]. In addition, CD1d−/− mice are universally deficient in the CD1d molecule and therefore unable to mediate iNKT cell interactions even after adoptive cell transfer [30]. We incubated

naïve wild-type iNKT cells with stimulatory lipids Selleck Cisplatin isolated from contact-sensitized mice, as shown earlier. We then transferred these activated iNKT cells into sensitized Jα18−/− and CD1d−/− mice and subsequently challenged their ears. CS was reconstituted in both strains, and the degree of reconstitution was nearly equivalent (Groups C and F, Fig. 4A). If endogenous iNKT cell interactions had been essential, then CS would have been seen in the Jα18−/− group but not in the CD1d−/− group. Rather, it appears that endogenous cellular interactions including hepatocyte–iNKT interactions are not essential. In our model, iNKT cell activation occurs at the in vitro stage in the context of other LMNC. Alternatively, in vivo cellular interactions involving different receptors may be at play.