NF-κB Is Activated in CD4(+) iNKT Cells by Sickle Cell Disease and Mediates Rapid Induction of Adenosine A(2A) Receptors

Reperfusion injury following tissue ischemia occurs as a consequence of vaso-occlusion that is initiated by activation of invariant natural killer T (iNKT) cells. Sickle cell disease (SDC) results in widely disseminated microvascular ischemia and reperfusion injury as a result of vaso-occlusion by rigid and adhesive sickle red blood cells. In mice, iNKT cell activation requires NF-κB signaling and can be inhibited by the activation of anti-inflammatory adenosine A(2A) receptors (A(2A)Rs). Human iNKT cells are divided into subsets of CD4+ and CD4- cells. In this study we found that human CD4+ iNKT cells, but not CD4- cells undergo rapid NF-κB activation (phosphorylation of NF-κB on p65) and induction of A(2A)Rs (detected with a monoclonal antibody 7F6-G5-A2) during SCD painful vaso-occlusive crises. These findings indicate that SCD primarily activates the CD4+ subset of iNKT cells. Activation of NF-κB and induction of A(2A)Rs is concordant, i.e. only CD4+ iNKT cells with activated NF-κB expressed high levels of A(2A)Rs. iNKT cells that are not activated during pVOC express low levels of A(2A)R immunoreactivity. These finding suggest that A(2A)R transcription may be induced in CD4+ iNKT cells as a result of NF-κB activation in SCD. In order to test this hypothesis further we examined cultured human iNKT cells. In cultured cells, blockade of NF-κB with Bay 11–7082 or IKK inhibitor VII prevented rapid induction of A(2A)R mRNA and protein upon iNKT activation. In conclusion, NF-κB-mediated induction of A(2A)Rs in iNKT cells may serve as a counter-regulatory mechanism to limit the extent and duration of inflammatory immune responses. As activated iNKT cells express high levels of A(2A)Rs following their activation, they may become highly sensitive to inhibition by A(2A)R agonists.