Injected versus Oral Cyclosporine for CD3 +, CD4 +, and CD8 + T-Cells in Wistar Rats
Objective: Cyclosporin-A (CsA) is a fungus-derived molecule that inhibits T-helper lymphocyte proliferation and has been largely used to downregulate cell-mediated immune responses during transplantation. CsA is widely employed clinically as immunosuppressive therapy in a variety of organ transplantations, as well as in the treatment of diseases of autoimmune origin. Moreover, CsA is frequently used in experimental transplantations in animal models. To test the hypothesis that may Low and high doses of CsA show any difference in lymphocyte (CD3+, CD4+, and CD8+ T-cells) percentage , we used an experimental model for 10, 20 and 40 mg/kg doses of oral cyclosporine regimens in Rat. Materials and Methods: Female rat of the outbred wistar stock, 6-8 weeks old, used in all of the experiments. Rats were randomized to two groups: 1. treatment and 2. control. The treatment group was administered 10 mg/kg per day dose CsA 1-3 weeks, 20 mg/kg 4-6 weeks and 40 mg/kg 6-9 weeks. Rats were weight and sacrificed weekly. The peripheral blood of rat collected in EDTA-tube and RBC cells lysed with Lysis buffer. Cells stained with CD3-FITC/CD4-PE/CD8-Alexa Flour647 antibodies (ab Serotec Company) and incubated in 4°C at 30 min. Samples were detected with FACS AriaII (BD Bioscience) at 488 nm and 633 nm laser extention. Data analyses waa done with FACS diva software. All of the samples collect to 20.000 events. Results: For statistical analyzes un-paired t-test was used to compare treated groups with their controls. At the end of the periods (3 weeks), there was no significant difference in T-cell count between different groups. Conclusion: CsA is an immunosuppressive drug that has been widely used to inhibit immune responses during transplant rejection. Result of this study demonstrates that Flow cytometry was not able to detect effect of CsA on decreasing immune activity when administered at multi doses. We recommend measure cytokine about immunosuppressive with ELISA test.