title: Evaluation of PLGA containing anti-CTLA4 inhibited endometriosis progression by regulating CD4+CD25+Treg cells in peritoneal fluid of mouse endometriosis model.
abstract: Our study investigated 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 particle size, zeta potential of the anti-CTLA-4 loaded nanoparticles was detected by dynamic light scattering. Morphology of nanoparticles was evaluated by transmission electron microscopy (TEM). Confocal laser scanning microscopy (CLSM) indicated distribution of anti-CTLA-4 with PLGA or without in peritoneal fluid. Cumulative anti-CTLA-4 release from nanoparticles was evaluated by Micro BCA assay. The percentage of CD4+CD25+Treg cells in peritoneal fluid was demonstrated by flow cytometer. 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 BrdU ELISA assay and Matrigel invasion assay. 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, 14days. 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. 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.
authors.affiliation: Department of Obstetrics & Gynecology, Graduate School of Tianjin Medical University, 22 Qixiangtai Road, Heping District, Tianjin 300070, China.
authors.forename: Qi
authors.initials: Q
authors.lastname: Liu
authors.affiliation: Department of Gynecology, Tianjin Central Hospital of Gynecology Obstetrics, 156 San Ma Road, Nankai District, Tianjin 300100,China.
authors.forename: Pingchuan
authors.initials: P
authors.lastname: Ma
authors.affiliation: Lab of Bioengineering, Institute of Biomedical Engineering, Chinese Academy of Medical Sciences & Peking Union Medical College, Tianjin Key Laboratory of Biomedical Materials, Tianjin 300192, China.
authors.forename: Lanxia
authors.initials: L
authors.lastname: Liu
authors.affiliation: Lab of Bioengineering, Institute of Biomedical Engineering, Chinese Academy of Medical Sciences & Peking Union Medical College, Tianjin Key Laboratory of Biomedical Materials, Tianjin 300192, China.
authors.forename: Guilei
authors.initials: G
authors.lastname: Ma
authors.affiliation: Lab of Bioengineering, Institute of Biomedical Engineering, Chinese Academy of Medical Sciences & Peking Union Medical College, Tianjin Key Laboratory of Biomedical Materials, Tianjin 300192, China.
authors.forename: Jingjing
authors.initials: J
authors.lastname: Ma
authors.affiliation: Lab of Bioengineering, Institute of Biomedical Engineering, Chinese Academy of Medical Sciences & Peking Union Medical College, Tianjin Key Laboratory of Biomedical Materials, Tianjin 300192, China.
authors.forename: Xiaoxuan
authors.initials: X
authors.lastname: Liu
authors.affiliation: Department of Obstetrics & Gynecology, Graduate School of Tianjin Medical University, 22 Qixiangtai Road, Heping District, Tianjin 300070, China.
authors.forename: Yijin
authors.initials: Y
authors.lastname: Liu
authors.affiliation: Department of Gynecology, Tianjin Central Hospital of Gynecology Obstetrics, 156 San Ma Road, Nankai District, Tianjin 300100,China.
authors.forename: Wanjun
authors.initials: W
authors.lastname: Lin
authors.affiliation: Department of Gynecology, Tianjin Central Hospital of Gynecology Obstetrics, 156 San Ma Road, Nankai District, Tianjin 300100,China. Electronic address: zhuyj8072@sina.com.
authors.forename: Yingjun
authors.initials: Y
authors.lastname: Zhu
mesh_terms: D000818:Animals; D000906:Antibodies / Q000627:therapeutic use*; D001202:Ascitic Fluid / Q000166:cytology; D060908:CTLA-4 Antigen / Q000037:antagonists & inhibitors*; D018929:Cell Culture Techniques; D049109:Cell Proliferation; D004337:Drug Carriers / Q000494:pharmacology; D004715:Endometriosis / Q000188:drug therapy* / Q000276:immunology; D005260:Female; D006801:Humans; D016753:Interleukin-10 / Q000378:metabolism; D019344:Lactic Acid / Q000737:chemistry*; D051379:Mice; D008810:Mice, Inbred C57BL; D046529:Microscopy, Electron, Transmission; D053758:Nanoparticles / Q000737:chemistry; D010316:Particle Size; D011100:Polyglycolic Acid / Q000737:chemistry*; D000077182:Polylactic Acid-Polyglycolic Acid Copolymer; D050378:T-Lymphocytes, Regulatory / Q000276:immunology*; D016212:Transforming Growth Factor beta / Q000378:metabolism
chemical_list: D000906:Antibodies; D060908:CTLA-4 Antigen; D004337:Drug Carriers; D016212:Transforming Growth Factor beta; D016753:Interleukin-10; D000077182:Polylactic Acid-Polyglycolic Acid Copolymer; D011100:Polyglycolic Acid; D019344:Lactic Acid
keywords: Anti-CTLA-4; CD4+CD25+Treg cells; Checkpoint inhibitors (CPI); Endometriosis model; PLGA