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Nephrol Dial Transplant (1994) 9: 1456-1461
© 1994 European Renal Association-European Dialysis and Transplant Association

research-article

In-vitro effects of cyclosporin A, FK506, 6-mercaptopurine, and prednisolone on lymphokine-activated killer cells

E. Alamartine1,, O. Sabido2 and F. Berthoux2,1

1Laboratoire de Néphrologie, Groupe de Recherche sur les Glomérulonéphrites et la Transplantation Rénale 2Center Commun de Cytométrie en Flux, Faculté de Médecine Jacques Lisfranc Saint Etienne, France

Correspondence and offprint requests to: Correspondence and offprint requests to: E. Alamartine, Service de Néphrologie Dialyse Transplantation Rénale, Hopital Nord, CHRU de Saint-Etienne, 42055 Saint Etienne Cedex 2, France

In transplant recipients, immunosuppressive regimens are deleterious on natural killer (NK) and lymphokine-activated killer (LAK) cells, which beyond their well-known antitumoral activity display many important biological functions. In order to find which regimens could preserve NK and LAK functions we tested the influence of CsA, FK506, 6-mercaptopurine and prednisolone on HLA-unrestricted cytotoxicity during an in-vitro IL-2 activation.

For each drug we obtained periphal blood samples from 11 healthy volunteers. Non-adherent PBMC were incubated 2 days with either CsA, FK506, 6-mercaptopurine or prednisolone, whose concentrations ranged from 0 to 10 µg/ml, in order to screen infratherapeutic, therapeutic, and supratherapeutic doses. Thereafter, 100 IU of IL-2 were added for a further 3-day culture. Before and after the culture, we analysed (1) the cell subsets by direct immunofluores cence staining with anti-CD3/CD16/CD56 antibodies, (2) the LAK activity with the lysis of Daudi cells, (3) the cell proliferation with a 24-h incorporation of thymidine.

Cyclosporin and FK506 did not impair the LAK cytotoxicity nor the number of LAK cells, whereas both prednisolone and 6-mercaptopurine decreased the LAK cytotoxicity, the number of CD3– CD16+ CD56+ cells, and the thymidine uptake. As a whole, the LAK cytotoxicity was correlated with the number of CD3– CD16+ CD56+ cells but not with the number of CD3– CD16– CD56– cells, and it also increased with the incorporation of thymidine. This latter was correlated with the number of CD3– CDl6+ CD56+ cells, but not with the number of CD3+ CD16– CD56– cells.

We conclude that in vitro the LAK activity is impaired by prednisolone and 6-mercaptopurine, but not by CsA or FK506, and that the deficiency of the LAK cytotoxicity seems to be related to the decreased number of IL-2-activated NK cells.

Keywords: CsA; FK506; 6-mercaptopurine; predisolone


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