Cell type-specific regulation of ITAM-mediated NF-κB activation by the adaptors, CARMA1 and CARD9

Activating NK cell receptors transduce signals through ITAM-containing adaptors, including FcR gamma and DAP12. Although the caspase recruitment domain (CARD)9-Bcl10 complex is essential for FcR gamma/DAP12-mediated NF-kappa B activation in myeloid cells, its involvement in NK cell receptor signaling is unknown. Herein we show that the deficiency of CARMA1 or Bcl10, but not CARD9, resulted in severe impairment of cytokine/chemokine production mediated by activating NK cell receptors due to a selective defect in NF-kappa B activation, whereas cytotoxicity mediated by the same receptors did not require CARMAI-BcI10-mediated signaling. licB kinase (IKK) activation by direct protein kinase C (PKC) stimulation with PMA plus ionomycin (P/I) was abrogated in CARMA1-deficient NK cells, similar to T and B lymphocytes, whereas CARD9-deficient dendritic cells (DCs) exhibited normal P/I-induced IKK activation. Surprisingly, CARMA1 deficiency also abrogated P/I-induced IKK activation in DCs, indicating that CARMA1 is essential for PKC-mediated NF-kappa B activation in all cell types, although the PKC-CARMA1 axis is not used downstream of myeloid ITAM receptors. Consistently, PKC inhibition abrogated ITAM receptor-mediated activation only in NK cells but not in DCs, suggesting PKC-CARMA1-independent, CARD9-dependent ITAM receptor signaling in myeloid cells. Conversely, the overexpression of CARD9 in CARNM1-deficient cells failed to restore the PKC-mediated NF-kappa B activation. Thus, NF-kappa B activation signaling through ITAM receptors is regulated by a cell type-specific mechanism depending on the usage of adaptors CARMA1 and CARD9, which determines the PKC dependence of the signaling.