ABSTRACT
Objective To investigate the effect of CKLF-like MARVEL transmembrane domain contai-ning member 5(CMTM 5)on EG-VEGF expression in prostate cancer cells , and to detect the potential mechanism of CMTM5 inhibiting prostate cancer .Methods The relative expression of CMTM 5 and EG-VEGF in prostate cells and prostate cancer cells was detected .Prostate cancer cells were given Plasmid transfection to overexpress CMTM5 and EG-VEGF expression was again detected .Then CMTM5 and EG-VEGF were compared between be-fore and after CMTM5 plasmid transfecting prostate cancer cells .Results Compared with the relative expresion of EG-VEGF and CMTM5 in prostate cells , prostate cancer cells showed high expression of EG-VEGF and low ex-pression of CMTM5, which were statistically significant , P<0.05.Compared with prostate cancer cells , the rela-tive expression of CMTM5 were obviously upregulated and EG-VEGF obviously decreased in prostate cancer cell after transfected by CMTM5 plasmid, which were statistically significant , P<0.05.Conclusions Prostate canc-er cells shows higher EG-VEGF expression and lower CMTM 5 campared with normal prostate cells .EG-VEGF is suppressed significantly when the prostate cancer cell is transfected by CMTM 5 plasmid and shows highlevel of CMTM5 expression , suggesting high expression of CMTM 5 may inhibit the development of prostate cancer by downregulating EG-VEGF expression .
ABSTRACT
OBJECTIVE@#To establish a method for in vitro expansion of human natural CD4⁺CD25⁺ T regulatory cell (Treg) cells for clinical study and immunotherapy.@*METHODS@#Human natural CD4⁺CD25⁺ Treg were isolated from peripheral blood monocyte cells (PBMCs) by magnetic activated cell sorting (MACS) and expanded by CD3/CD28 expansion beads, IL-2 and rapamycin. The number and the viability of the freshly isolated and expanded Treg were detemined by trypan blue staining. The phenotype and the purity of the freshly isolated and expanded Treg were analyzed by FACS. Treg suppression activity was assessed by mixed lymphocyte reaction (MLR) assay.@*RESULTS@#Human natural Treg were expanded up to 2 000 folds after 3 weeks in culture, and the activity was more than 97%. The expanded Treg retained Treg phenotype as shown by their freshly isolated counterparts, and the purity of CD4⁺CD25⁺FoxP3⁺ Treg was (94.22 ± 2.12)%. The expanded Treg demonstrated a similar potent suppression of both proliferating auto- and allo- CD4⁺CD25⁻ effector T cells in vitro in a cell number-dependent manner.@*CONCLUSION@#An in vitro expansion of human natural Treg was established to obtain large numbers of human Treg with highly suppressive phenotype and function, thereby providing a solution to the availability of sufficient human natural Treg in clinical study and immunotherapy.
Subject(s)
Humans , Cell Culture Techniques , Cell Separation , Cells, Cultured , Interleukin-2 , Leukocytes, Mononuclear , Lymphocyte Culture Test, Mixed , T-Lymphocytes, Regulatory , Cell BiologyABSTRACT
Objective To study the role of glucagon-like peptide-1 (GLP-1) analogue liraglutide played in the proliferation of CD4+CD25 T cells in normal people and newly-onset type 1 diabetic patients,and to evaluate the possible immune regulatory role of liraglutide in the therapy of type 1 diabetes.Methods CD4+ CD25-T cells of 10 normal people and 10 newly-onset type 1 diabetic patients were separated from peripheral blood by MACS immunomagnetic beads and stimulated by Human T-Activator CD3/CD28 Dynabeads to proliferate.CFSE labeling technique was used to evaluate the proliferation of CD4+ CD25-T cells by flow cytometry.Liraglutide of different concentrations(0,25,50,and 100 nmol/ml) was added to the proliferation system,then the proliferation of CD4+CD25-T cell was measured.Results (1) Liraglutide suppressed the proliferation of CD4+ CD25-T cells from either normal people or type 1 diahetic patients with dose-dependent manner (P ＜ 0.05).(2) Under the different concentrationsofliraglutide,the proliferation ofCD4+CD25 T cells from diabetic patients was mueh more robust than that of normal people (P＜0.01).(3) The inhibitory effects of liraglutide on CD4+ CD25-T cells proliferation in normal people and diabetic patients were similar (P＞0.05).Conclusion The proliferation of CD4+ CD25 T cells in type 1 diabetic patients was more robust than normal people,which indicated cellular immune dysfunction in type 1diabetes.Liraglutide inhibits the proliferation of CD4+ CD25-T cells of type 1 diabetic patients in vitro.The immunosuppression effect of liraglutide may have potential value in the treatment of type 1 diabetes.