Assessment of the inhibitory effect of immunosuppressive agents on rat T cell interferon-γ production using an ELISPOT assay
article
The immunospot (ELISPOT) assay has proven to be an efficient and sensitive method for the enumeration of single cells secreting antibodies or cytokines. Here we show that the generation of interferon-γ producing cells (IFN-γ pc) in rat spleen cell cultures stimulated with concanavalin A (ConA) is dose-dependently inhibited by a wide variety of immunosuppressants such as cyclosporin A, FK506, hydrocortisone, dexamethasone, azathioprine and ART-18, a monoclonal antibody (mAb) with established immunosuppressive activity in organ transplantation and autoimmunity. The minimal inhibitory concentration (m.i.c.) correlated well with the reported m.i.c. in the mixed lymphocyte reaction (MLR) assay and the therapeutically effective plasma levels in vivo. Our data indicate that the IFN-γ-specific immunospot assay is a powerful tool for determining the potency of immunosuppressive agents in vitro and provides a simple and accurate method for screening large numbers of agents with suspected immunosuppressive properties. The assay may additionally prove to be of value for determining the therapeutically effective doses of immunosuppressants that should be administered in vivo. Chemicals/CAS: Antibodies, Monoclonal; Histocompatibility Antigens Class II; Immunosuppressive Agents; Interferon Type II, 82115-62-6; Interleukin-2; Methotrexate, 59-05-2
Topics
concanavalin AELISPOT assayimmunosupressantsinterferon-γsplenocyteazathioprinecyclosporin Agamma interferonhydrocortisoneimmunosuppressive agentinterleukin 2methotrexatetacrolimusanimal cellanimal experimentarticlefemaleimmunoassaynonhumanpriority journalratspleen cellT lymphocyteAnimalAntibodies, MonoclonalCells, CulturedFemaleHistocompatibility Antigens Class IIImmunosuppressive AgentsInterferon Type IIInterleukin-2MethotrexateRatsRats, Inbred StrainsSupport, Non-U.S. Gov'tT-LymphocytesAnimalia
TNO Identifier
231628
DOI
https://dx.doi.org/10.1016/0022-1759(91)90087-V
ISSN
00221759
Source
Journal of Immunological Methods, 144(2), pp. 203-213.
Pages
203-213
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