Evaluation Of PLGA/anti-CTLA-4 Suppressive Effect On Mouse Endometriosis Model

Objective: Our study in order to investigated whether poly(lactic-co-glycolic acid)(PLGA)as protein delivery vehicles encapsulate CTLA-4-antibody(anti-CTLA-4)which is essential for CD4+CD25+Treg cells suppressive function exposing superior potential for inhibiting endometriosis progress in mouse model than single anti-CTLA-4.Anti-CTLA-4 loaded PLGA combined to ligands CTLA-4 in surface of CD4+CD25+Treg cells which distributed in peritoneal fluid of mouse endometriosis model.The effect of PLGA/anti-CTLA-4 on endometriosis mouse model.Method:1.The particle size,zeta potential of the anti-CTLA-4 loaded nanoparticles was detected by dynamic light scattering.2 Morphology of nanoparticles was evaluated by transmission electron microscopy(TEM).3.Confocal laser scanning microscopy(CLSM)indicated distribution of anti-CTLA-4 with PLGA or without in peritoneal fluid 4.Cumulative anti-CTLA-4 release from nanoparticles was evaluated by Micro BCA assay.5 The percentage of CD4+CD25+Treg cells in peritoneal fluid was demonstrated by flow cytometer.6 In vitro experiment we co-culture ectopic endometrial cells(EEC)with isolated CD4+CD25+Treg cells in peritoneal fluid(PF),proliferation and invasion of ectopic endometrial cells(EEC)was measured by Brd U ELISA assay and Matrigel invasion assay.7.In comparison with anti-CTLA-4 without nanoparticles,the bioconjugates PLGA/anti-CTLA-4 were tolerated in peritoneal fluid with a controlled release of anti-CTLA-4 in 3,7,14 days.8 Moreover,PLGA/anti-CTLA-4 had superior protective regulation ability to reduce level of CD4+CD25+Treg cells in peritoneal fluid.Most strikingly,in vitro experiment,PLGA/anti-CTLA-4 exhibited better ability in inhibiting proliferation and invasion of ectopic endometrial cells in co-culture system compared with anti-CTLA-4.9.IL-10 and TGF-βproduction secreted by CD4+CD25+Treg cells in peritoneal fluid is assessed by ELISA.Conclusion:Progressively,PLGA/anti-CTLA-4 had better suppressive activity to inhibited IL-10 and TGF-beta secreted by CD4+CD25+Treg cells which indicating that PLGA/anti-CTLA-4 suppressed cells proliferation and invasion through reduced IL-10 and TGF-beta production.Thus,PLGA/anti-CTLA-4 may be a potential strategy for endometriosis therapy.