title: IL-21 and IL-12 inhibit differentiation of Treg and TH17 cells and enhance cytotoxicity of peripheral blood mononuclear cells in patients with cervical cancer.
abstract: OBJECTIVE
  Interleukin 21 (IL-21) and IL-12 have been known to be effective antitumor agents. In this study, we evaluated whether IL-21 in combination with IL-12 could enhance the cytotoxicity of peripheral blood mononuclear cells (PBMCs) in patients with cervical intraepithelial neoplasia III and cervical cancer.
  
  
  METHODS
  Peripheral blood mononuclear cells were isolated from peripheral blood of cervical intraepithelial neoplasia III patients (n = 17) and cervical cancer patients (n = 18). Peripheral blood mononuclear cells were cultured with IL-2 in low concentration as control group. Interleukin 2-stimulated PBMCs were cocultured with anti-human IL-21 neutralizing antibody, IL-21 alone, IL-12 alone, and IL-21 plus IL-12, respectively, as test groups. The cytotoxicity of PBMCs against SiHa tumor cells was examined by lactate dehydrogenase released assay. CD4CD25FOXP3 T regulatory (Treg) cells and CD4IL-17A T helper 17 (TH17) cells were analyzed by flow cytometry. Proliferation and apoptosis were detected by CCK-8 (cell counting kit 8) assay and flow cytometry, respectively.
  
  
  RESULTS
  Compared with controls, IL-21 and IL-12 significantly elevated PBMC cytotoxicity against SiHa cells. Moreover, IL-21 plus IL-12 significantly elevated PBMC cytotoxicity in comparison to IL-21 alone and IL-12 alone. We also found that IL-21 plus IL-12 significantly decreased Treg and TH17 cell proportion in comparison to controls. Notably, IL-21 plus IL-12 significantly decreased TH17 cell proportion in comparison to IL-21 alone. Both IL-21 and IL-12 significantly decreased the apoptosis rate of PBMCs, whereas neither IL-21 nor IL-12 had significant effect on PBMC proliferation.
  
  
  CONCLUSIONS
  The combination of IL-21 and IL-12 could efficiently stimulate PBMCs with cytotoxicity against SiHa cells, and the possible mechanisms may be due to down-regulated Treg and TH17 cell differentiation.
authors.affiliation: Department of Obstetrics and Gynecology; Qilu Hospital, Shandong University, Shandong Cancer Hospital & Institute, Ji'nan, People's Republic of China.
authors.forename: Yongju
authors.initials: Y
authors.lastname: Tian
authors.forename: Cunzhong
authors.initials: C
authors.lastname: Yuan
authors.forename: Daoxin
authors.initials: D
authors.lastname: Ma
authors.forename: Yan
authors.initials: Y
authors.lastname: Zhang
authors.forename: Yi
authors.initials: Y
authors.lastname: Liu
authors.forename: Wenjing
authors.initials: W
authors.lastname: Zhang
authors.forename: Fei
authors.initials: F
authors.lastname: Hou
authors.forename: Baoxia
authors.initials: B
authors.lastname: Cui
mesh_terms: D000328:Adult; D016022:Case-Control Studies; D002454:Cell Differentiation / Q000187:drug effects / Q000276:immunology; D045744:Cell Line, Tumor; D005260:Female; D006801:Humans; D018664:Interleukin-12 / Q000276:immunology / Q000494:pharmacology*; D007378:Interleukins / Q000276:immunology / Q000494:pharmacology*; D007963:Leukocytes, Mononuclear / Q000187:drug effects* / Q000276:immunology; D050378:T-Lymphocytes, Regulatory / Q000187:drug effects* / Q000276:immunology; D058504:Th17 Cells / Q000187:drug effects* / Q000276:immunology; D002583:Uterine Cervical Neoplasms / Q000097:blood / Q000188:drug therapy / Q000276:immunology*; D002578:Uterine Cervical Dysplasia / Q000097:blood / Q000276:immunology; D000098546:Interleukin-21
chemical_list: D007378:Interleukins; D018664:Interleukin-12; D000098546:Interleukin-21