A calcium optimum for cytotoxic T lymphocyte and natural killer cell cytotoxicity

Key points: Cytotoxic T lymphocytes (CTLs) and natural killer (NK) cells are required to eliminate cancer cells. We analysed the Ca²⁺ dependence of CTL and NK cell cytotoxicity and found that in particular CTLs have a very low optimum of [Ca²⁺ ]_i (between 122 and 334 nm) and [Ca²⁺ ]_o (between 23 and 625 μm) for efficient cancer cell elimination, well below blood plasma Ca²⁺ levels. As predicted from these results, partial down-regulation of the Ca²⁺ channel Orai1 in CTLs paradoxically increases perforin-dependent cancer cell killing. Lytic granule release at the immune synapse between CTLs and cancer cells has a Ca²⁺ optimum compatible with this low Ca²⁺ optimum for efficient cancer cell killing, whereas the Ca²⁺ optimum for CTL migration is slightly higher and proliferation increases monotonously with increasing [Ca²⁺ ]_o . We propose that a partial inhibition of Ca²⁺ signals by specific Orai1 blockers at submaximal concentrations could contribute to tumour elimination.

Abstract: Cytotoxic T lymphocytes (CTLs) and natural killer (NK) cells are required to protect the human body against cancer. Ca²⁺ is a key metabolic factor for lymphocyte function and cancer homeostasis. We analysed the Ca²⁺ dependence of CTL and NK cell cytotoxicity against cancer cells and found that CTLs have a bell-shaped Ca²⁺ dependence with an optimum for cancer cell elimination at rather low [Ca²⁺ ]_o (23-625 μm) and [Ca²⁺ ]_i (122-334 nm). This finding predicts that a partial inhibition of Orai1 should increase (rather than decrease) cytotoxicity of CTLs at [Ca²⁺ ]_o higher than 625 μm. We tested this hypothesis in CTLs and indeed found that partial down-regulation of Orai1 by siRNA increases the efficiency of cancer cell killing. We found two mechanisms that may account for the Ca²⁺ optimum of cancer cell killing: (1) migration velocity and persistence have a moderate optimum between 500 and 1000 μm [Ca²⁺ ]_o in CTLs, and (2) lytic granule release at the immune synapse between CTLs and cancer cells is increased at 146 μm compared to 3 or 800 μm, compatible with the Ca²⁺ optimum for cancer cell killing. It has been demonstrated in many cancer cell types that Orai1-dependent Ca²⁺ signals enhance proliferation. We propose that a decrease of [Ca²⁺ ]_o or partial inhibition of Orai1 activity by selective blockers in the tumour microenvironment could efficiently reduce cancer growth by simultaneously increasing CTL and NK cell cytotoxicity and decreasing cancer cell proliferation.