ABSTRACT
OBJECTIVE@#To investigate the effects of exosomes from human umbilical cord mesenchymal stem cells on the development of Treg and TH17 cells.@*METHODS@#Exosomes from the serum-free-culture supernatants of hUC-MSC were harvested by ultracentrifugation. The electron microscopy, nanoparticle tracking analysis and western blot were used to identify the hUC-MSC-exosomes, such as the morphology, the paticle chameter, and the protein content. The PBMC stimulated with anti-CD3/CD28 were incubated with the exosomes for five days, and then the percentage changes of Treg and TH17 cells were analyzed by using flow cytometry.@*RESULTS@#The hUC MSC-derived exosomes were saucer-like in morphology the averge diameter was approximately 142 nm. They were identified as positive for CD9 and CD63. Flow cytometry showed that the proportion of CD4CD25Foxp3 Treg cells in the PBMC were significantly higher, but the proportion of CD4IL17A T cells in the hUC-MSC-exosome group was obviously lower than that in the group without the hUC-MSC-exosom (control group) (P<0.05).@*CONCLUSION@#The hUC-MSC-exosomes have an immunomodulatory effect on T cells in vitro by increasing the ratio of Treg and reducing the ratio of TH17 cells, expecting the hUC-MSC-exosom as a novel cell-free target for immunotherapy.
Subject(s)
Humans , Exosomes , Leukocytes, Mononuclear , Mesenchymal Stem Cells , T-Lymphocytes, Regulatory , Th17 Cells , Umbilical CordABSTRACT
Purpose To investigate the effect of DAPPER1 on the malignant biological behavior in esophageal carcinoma cells,and further to analyze the expression and the possible regulated mechanism of DAPPER1 in esophageal squamous cell cancer (ESCC) samples.Methods MTT assay,colony formation assay and wound healing were used to examine the effect of DAPPER1 on malignant biological behavior in esophageal carcinoma cells,RT-PCR and MSP methods were applied respectively to examine the expression and the methylation status of DAP-PER1 in three ESCC cell lines (TE13,T.Tn,Eca109).Immunohistochemistry was used to examine the DAPPER1 protein expression.Results The negative or weak expression of DAP-PER1 was detected in ESCC cell lines.After treated with 5-Aza2'-deoxycytidine (5-Aza-dC,a demethylation agent),the expression level of DAPPER1 was obviously increased.Meanwhile,over expression of DAPPER1 or treated with 5-Aza-Dc could obviously inhibit proliferation and immigration abilities of TEl3 cell line.The level of DAPPER1 expression had no obviously change after treated with trichostatin A (TSA).Decreased protein expression of DAPPER1 was observed in ESCC tumor tissues compared with non-cancerous tissues (P ＜ 0.01),and associated with the methylation status of this gene (P ＜ 0.01).Conclusion DAPPER1 plays an important role of tumor suppressor gene in esophageal carcinoma,and the aberrant hypermethylation may be one of the main mechanisms in abnormal expression of this gene.