Impact of Commonly Used Transplant Immunosuppressive Drugs on Human NK Cell Function Is Dependent upon Stimulation Condition

Abstract

Lung transplantation is a recognised treatment for patients with end stage pulmonary disease. Transplant recipients receive life-long administration of immunosuppressive drugs that target T cell mediated graft rejection. However little is known of the impact on NK cells, which have the potential to be alloreactive in response to HLA-mismatched ligands on the lung allograft and in doing so, may impact negatively on allograft survival. NK cells from 20 healthy controls were assessed in response to Cyclosporine A, Mycophenolic acid (MPA; active form of Mycophenolate mofetil) and Prednisolone at a range of concentrations. The impact of these clinically used immunosuppressive drugs on cytotoxicity (measured by CD107a expression), IFN-γ production and CFSE proliferation was assessed in response to various stimuli including MHC class-I negative cell lines, IL-2/IL-12 cytokines and PMA/Ionomycin. Treatment with MPA and Prednisolone revealed significantly reduced CD107a expression in response to cell line stimulation. In comparison, addition of MPA and Cyclosporine A displayed reduced CD107a expression and IFN-γ production following PMA/Ionomycin stimulation. Diminished proliferation was observed in response to treatment with each drug. Additional functional inhibitors (LY294002, PD98059, Rottlerin, Rapamycin) were used to elucidate intracellular pathways of NK cell activation in response to stimulation with K562 or PMA-I. CD107a expression was significantly decreased with the addition of PD98059 following K562 stimulation. Similarly, CD107a expression significantly decreased following PMA-I stimulation with the addition of LY294002, PD98059 and Rottlerin. Ten lung transplant patients, not receiving immunosuppressive drugs pre-transplant, were assessed for longitudinal changes post-transplant in relation to the administration of immunosuppressive drugs. Individual patient dynamics revealed different longitudinal patterns of NK cell function post-transplantation. These results provide mechanistic insights into pathways of NK cell activation and show commonly administered transplant immunosuppression agents and clinical rejection/infection events have differential effects on NK cell function that may impact the immune response following lung transplantation.

Improved outcome of children transplanted for high-risk leukemia by using a new strategy of cyclosporine-based GVHD prophylaxis

“…Indeed, CsA pharmacodynamic effect on NK cells is distinct and allows NK cell cytotoxicity, 41 whereas it is impaired by MMF [42][43][44] or MTX. [45][46][47] Moreover, calcineurin inhibitors are the only immunosuppressive agents exerting their action by inhibiting interleukin-2 production.…”

A phase I/II trial of interleukin-15–stimulated natural killer cell infusion after haplo-identical stem cell transplantation for pediatric refractory solid tumors

“…However, there are contradictory reports in the literature [45e47]. It can be suggested that NK cell stimulation occurs through multiple pathways of intracellular activation (including protein kinase C and calcineurin pathways) [48]. The potent inhibitory effects of mycophenolate mofetil (MMF) on NK cell proliferation and cytotoxic activity are apparently more evident than those of other immunosuppressants, including CsA, tacrolimus and methotrexate [49] The NK-activating receptor expression of NK cells is downregulated by MMF, and the inhibitory effect is partially abrogated.…”

PMA and Ionomycin Induce Glioblastoma Cell Death: Activation-Induced Cell-Death-Like Phenomena Occur in Glioma Cells

“…PMA and Io are routinely used to establish a T cell receptor-independent model to study T cell activation and proliferation [14]. By mimicking the phospholipase C-driven activation of PKC and the increase of cytosolic Ca 2+ , PMA and Io activate the transcription factors NFAT1, NF-κB and activator protein-1 (AP-1), and subsequently regulate downstream gene expression [38].…”

“…Therefore, we speculate that factors activating NFAT1, such as phorbol myristate acetate (PMA) and ionomycin (Io), will further influence GBM cell growth. The combination of PMA and Io has been widely used in the study of T cell activation [11], [12], [13], [14]. Through activation of protein kinase C (PKC) and calcineurin, PMA/Io can activate many transcription factors, including NF-κB and members of the NFAT family, and subsequently regulate the expression of numerous genes [13].…”