Inhibition of alloantigen presentation by cyclosporine.
Cyclosporine (CsA) was examined for its ability to inhibit alloantigen presentation by spleen cells in a primary mixed lymphocyte reaction; by gamma interferon-induced P388.D1 macrophages to an alloreactive T cell clone; and by a B cell lymphoma, B1D.beta to an alloreactive T cell hybridoma. Alloantigen-presenting cells were treated with CsA or its inactive analogs for 2 hr, washed extensively (four times), and added to the T cells. Using this protocol, CsA maximally inhibited allorecognition by the T cells at 1000 ng/ml in all three systems. An HPLC assay for CsA cell failed to detect any significant CsA carryover into the T cell assays. Supernatant transfer experiments also failed to demonstrate CsA carryover in the more sensitive T cell hybridoma assay. These transfer experiments also failed to demonstrate the generation of inhibitory factors during the assay. Northern blot analysis and a cell-surface ELISA failed to observe any decreases in MHC class II induction in/on P388.D1 cells with CsA present during the induction. Due to the lack of detectable (less than 10 ng/ml) CsA carryover, we hypothesize that CsA has a direct effect on the formation of stimulatory MHC class II in our alloreactive systems.
Little
R G
RG
Department of Immunology, Mayo Clinic, Rochester, Minnesota 55905.
Ebertowski
L A
LA
David
C S
CS
D000818:Animals; D000938:Antigen-Presenting Cells / Q000276:immunology*; D015152:Blotting, Northern; D002460:Cell Line; D003524:Cyclosporins / Q000494:pharmacology*; D004305:Dose-Response Relationship, Drug; D000949:Histocompatibility Antigens Class II / Q000235:genetics / Q000276:immunology; D066298:In Vitro Techniques; D008213:Lymphocyte Activation / Q000187:drug effects; D007959:Lymphocyte Culture Test, Mixed; D008264:Macrophages / Q000502:physiology; D051379:Mice; D008815:Mice, Inbred Strains; D012333:RNA, Messenger / Q000235:genetics
D003524:Cyclosporins; D000949:Histocompatibility Antigens Class II; D012333:RNA, Messenger