Effects Of Rapamycin On Immunophenotype Of Dendritic Cells: A Preliminary Study

Objective:Test if rapacymin has the impact on the change of immunosuppressive agents and level of secretion of cytokine. Study the physical change of dentritic cells from immature to mature This study can produce experimental data to support the later study of the tolerance of dentritic cells during organ transplant.Methods:Took the tibia and femur off from 6 to 8 week old male SD mice.In sterile conditions, washed out and cleaned bone marrow cells, then filtered to remove some of the red cells by means of ficoll method, the cells obtained are in 10% fetal bovine. Developed in GM-CSF(10ng/ml)and IL4 (5ng/ml) RPMI1640 cultures, Change half of the liquid every other day.Added GM-CSF、IL-4 in the same concentration, meanwhile observing the change of the appearance of the dentritic cells using optical microscope.On the fourth day of development, added 5mg/ml of rapacymin into Group A, added 30mg/ml of rapacymin into group B,Added 60mg/ml of rapacymin into group C, D kept untreated.24 hours later, collected the supernatant to test the level secrection of IL-2、IL-6、IL-10 and IL-12 produced from 4 groups.Collected the cells by flow cytometry, then tested the differences among the surface of dentritic cells CD80、CD86、CD103 and MH-Ⅱ.Reasult:1. Rapacymin has impact on the level of secretion of dentritic cytokine: collected the supernatart which has been developed for 7 days. Tested the how much cytokine enzyme produced from 4 groups by immunoassays method.However the data shows different amount of rapacymin has different impact on the cykotine production of dentritic cells, it depends on the quantity of rapacymin.In this experiment,5ng/ml and30ng/ml has more obvious impact on it.2. The impact of rapacymin on the immunosuppressive agents of dentritic cells:on the seventh development day, after collecting all groups of dentritic cells, using flow cytometer to test the expression of the surface molecules of cells, results showed that rapacimin has impact on treated dentritic cells expression. Among all of the results, the expression of CD80、CD86、MH-Ⅱhas obviously become higher, but the expression of CD103 has no obvious change. The expression of group of 30ng/ml has the most obvious change.Conclusion:1. Rapacimin has the impact on the level of production of the cytokine and surface molecules immunosuppressive agents.2. rapacymin in different concentrations may have different impact on the production of the cytokine and surface molecules immunosuppressive agents.3. Rapacymin may interfere with the ripening process of dendritic cells.