Detection and quantification of cyclosporine in body fluids using an interleukin-2 reporter-gene assay.
Different assays are employed to monitor the concentration of immunosuppressive drugs in biological fluids. None of these methods gives direct and precise information on the actual level of immunosuppression in the patient. Here we describe the use of an interleukin-2 (IL-2) reporter-gene assay (IL-2 RGA) to monitor the concentrations of immunosuppressants in body fluids. This assay is based on a chimeric gene construct in which the human IL-2 promoter drives the expression of a reporter gene. Upon mitogenic stimulation the reporter gene is expressed and can be easily quantified. The assay is very sensitive and selective for immunosuppressive compounds inhibiting IL-2 gene expression such as cyclosporine (CsA) and FK506, their active metabolites and derivatives, but not for others such as rapamycin. High reproducibility, fast performance time, and high capacity are additional characteristics of the assay. The assay was developed to monitor immunosuppressive drug levels in human volunteers or in animals receiving CsA analogues as the only immunosuppressive drugs. This assay is sensitive to CsA or ascomycin/FK506 analogues and metabolites, for which there are presently no specific monoclonal antibodies available. The IL-2 reporter-gene assay may be more suitable than other in vitro systems such as MLR or mitogen stimulated PBMC which were previously used to study the immunosuppressive activity of drugs in body fluids.
Wenner
P R
PR
Preclinical Research, Sandoz Pharma Ltd., Basel, Switzerland.
Di Padova
F
F
Keown
P A
PA
D000591:Amino Acid Isomerases / Q000378:metabolism; D001681:Biological Assay; D002352:Carrier Proteins / Q000378:metabolism; D002460:Cell Line; D016572:Cyclosporine / Q000097:blood* / Q000378:metabolism; D015870:Gene Expression / Q000187:drug effects; D017930:Genes, Reporter; D006801:Humans; D007165:Immunosuppression Therapy; D007166:Immunosuppressive Agents / Q000032:analysis; D007376:Interleukin-2 / Q000235:genetics; D019696:Peptidylprolyl Isomerase; D011401:Promoter Regions, Genetic; D011485:Protein Binding
D002352:Carrier Proteins; D007166:Immunosuppressive Agents; D007376:Interleukin-2; D016572:Cyclosporine; D000591:Amino Acid Isomerases; D019696:Peptidylprolyl Isomerase