Limited efficacy of immunosuppressive drugs on CD8+ T cell-mediated and natural killer cell-mediated lysis of human renal tubular epithelial cells.
BACKGROUND
Although CD8+ T cell-mediated and natural killer (NK) cell-mediated cytotoxicity against renal tubular epithelial cells (TECs) plays a crucial role during rejection, the degree of inhibition of these lytic immune responses by immunosuppressive drugs is unknown. We investigated the CD8 T-cell and NK cell responses induced by TECs in vitro and questioned how these processes are affected by immunosuppressive drugs.


METHODS
Donor-derived TECs were co-cultured with recipient peripheral blood monocyte cells. Proliferation of CD8+ T cells and NK cell subsets was assessed using PKH dilution assay. CD107a degranulation and europium release assay were performed to explore CD8+-mediated and NK cell-mediated TEC lysis. Experiments were conducted in the absence or presence of tacrolimus (10 ng/mL), everolimus (10 ng/mL), and prednisolone (200 ng/mL).


RESULTS
Tubular epithelial cells induce significant CD8+ T-cell and NK cell proliferation. All immunosuppressive drugs significantly inhibited TEC-induced CD8+ T-cell proliferation. Interestingly, prednisolone was the most powerful inhibitor of NK cell proliferation. CD8-mediated and NK cell-mediated early lytic responses were marked by strong degranulation after an encounter of unstimulated TECs, represented by a high cell surface expression of CD107a. However, with the use of interferon-γ-activated and tumor necrosis factor-α-activated TECs, the NK degranulation response was significantly reduced and CD8 degranulation response was even more enhanced (P<0.05). Tubular epithelial cell-induced CD8 degranulation and CD8-mediated TEC lysis were preferentially inhibited by tacrolimus and prednisolone, and not by everolimus. Although tacrolimus showed the most inhibitory effect on the degranulation of NK cells, NK cell-mediated TEC lysis was efficiently inhibited by prednisolone (P<0.05).


CONCLUSIONS
Overall, our data point to a limited efficacy of immunosuppressive drugs on CD8+ T cell-mediated and NK cell-mediated lysis of human renal TECs.
1 Department of Internal Medicine, Division of Nephrology and Transplantation, Erasmus MC, Division of Nephrology and Transplantation, University Medical Center Rotterdam, the Netherlands. 2 Address correspondence to: Martijn Demmers, M.S.c., Division of Nephrology and Transplantation, Department of Internal Medicine, Division of Nephrology and Transplantation, Erasmus MC, University Medical Center Rotterdam, PO Box 2040, 3000 CA Rotterdam, the Netherlands; Ajda T. Rowshani, M.D., Ph.D., Department of Internal Medicine, Division of Nephrology and Transplantation, Erasmus MC, University Medical Center Rotterdam, PO Box 2040, 3000 CA Rotterdam, the Netherlands.
Martijn W H J
MW
Demmers
Sander S
SS
Korevaar
Michiel G H
MG
Betjes
Willem
W
Weimar
Ajda T
AT
Rowshani
Carla C
CC
Baan
D018414:CD8-Positive T-Lymphocytes / Q000187:drug effects*; D049109:Cell Proliferation; D016207:Cytokines / Q000378:metabolism; D004847:Epithelial Cells / Q000187:drug effects*; D005063:Europium / Q000737:chemistry; D000068338:Everolimus; D006801:Humans; D007166:Immunosuppressive Agents / Q000494:pharmacology*; D007249:Inflammation; D007684:Kidney Tubules / Q000187:drug effects*; D007694:Killer Cells, Natural / Q000187:drug effects*; D052118:Lysosomal-Associated Membrane Protein 1 / Q000378:metabolism; D009000:Monocytes / Q000166:cytology / Q000187:drug effects; D011239:Prednisolone / Q000494:pharmacology; D020123:Sirolimus / Q000031:analogs & derivatives / Q000494:pharmacology; D016559:Tacrolimus / Q000494:pharmacology
D016207:Cytokines; D007166:Immunosuppressive Agents; D052118:Lysosomal-Associated Membrane Protein 1; D005063:Europium; D000068338:Everolimus; D011239:Prednisolone; D020123:Sirolimus; D016559:Tacrolimus