Common bile duct(CBD)stones are a frequent problem in Chinese populations,and their incidence is particularly high in certain areas(Wang et al.,2013).In recent years,laparoscopic common bile duct exploration(LCBDE)and...Common bile duct(CBD)stones are a frequent problem in Chinese populations,and their incidence is particularly high in certain areas(Wang et al.,2013).In recent years,laparoscopic common bile duct exploration(LCBDE)and endoscopic retrograde cholangiopancreatography(ERCP)have been the main surgical procedures for CBD stones,although each has different advantages and disadvantages in the treatment of choledocholithiasis(Loor et al.,2017;Zhou et al.,2017).展开更多
Objective： The aim of our study is to observe the impact of cholangiocarcinoma-derived exosomes on the antitumor activities of cytokine-induced killer （CIK） cells and then demonstrate the appropriate mechanism. Met...Objective： The aim of our study is to observe the impact of cholangiocarcinoma-derived exosomes on the antitumor activities of cytokine-induced killer （CIK） cells and then demonstrate the appropriate mechanism. Methods： Tumor-derived exosomes （TEXs）, which are derived from RBE cells （human cholangiocarcinoma line）, were collected by ultracentrifugation. CIK cells induced from peripheral blood were stimulated by TEXs. Fluorescence-activated cell sorting （FACS） was performed to determine the phenotypes of TEX-CIK and N-CIK （normal CIK） cells. The concen- trations of tumor necrosis factor-a （TNF-a） and perforin in the culture medium supematant were examined by using an enzyme-linked immunosorbent assay （ELISA） kit. A CCK-8 kit was used to evaluate the cytotoxic activity of the CIK cells to the RBE cell line. Results： The concentrations of TNF-a and perforin of the group TEX-CIK were 138.61 pg/ml and 2.41 ng/ml, respectively, lower than those of the group N-CIK 194.08 pg/ml （P〈0.01） and 3.39 ng/ml （P〈0.05）. The killing rate of the group TEX-CIK was 33.35%, lower than that of the group N-CIK （47,35% （P〈0.01））. The population of CD3＋, CD8＋, NK （CD56＋）, and CD3＋CD56＋ cells decreased in the TEX-CIK group （63.2±6.8）%, （2.5±1.0）%, （0.53±0.49）%, （0.45±0.42）%） compared with the N-CIK group （（90.3±7.3）%, （65.7±3.3）%, （4.2±1.2）%, （15.2±2.7）%）, P〈0.01. Conclusions： Our results suggest that RBE cells-derived exosomes inhibit the antitumor activity of CIK cells by down-regulating the population of CD3＋, CD8＋, NK （CD56＋）, and CD3＋CD56＋ cells and the secretion of TNF-a and perforin. TEX may play an important role in cholangiocarcinoma immune escape.展开更多
Background： Tumor-derived exosomes were considered to be potential candidates for tumor vaccines because they are abundant in immune-regulating proteins, whereas tumor exosomal miRNAs may induce immune tolerance, the...Background： Tumor-derived exosomes were considered to be potential candidates for tumor vaccines because they are abundant in immune-regulating proteins, whereas tumor exosomal miRNAs may induce immune tolerance, thereby having an opposite immune function. Objective： This study was designed to separate exosomal protein and depleted exosomal microRNAs （miRNAs）, increasing the immune activity of exosomes for activating dendritic cell/cytokine-induced killer cells （DC/CIKs） against pancreatic cancer （PC）. Methods： PC-derived exosomes （PEs） were extracted from cultured PANC-1 cell supernatants and then ruptured; this was followed by ultrafiltered exosome lysates （UELs）. DCs were stimulated with lipopolysaccharide （LPS）, PE, and UEL, followed by co-culture with CIKs. The anti-tumor effects of DC/CIKs against PC were evaluated by proliferation and killing rates, tumor ne- crosis factor-a （TNF-a） and perforin secretion. Exosomal miRNAs were depleted after lysis and ultrafiltration, while 128 proteins were retained, including several immune-activating proteins. Results： UEL-stimulated DC/CIKs showed a higher killing rate than LPS- and PE-stimulated DC/CIKs. Conclusions： miRNA-depleted exosome proteins may be promising agonists for specifically activating DC/CIKs against PC.展开更多
基金Project supported by the Foundation Project for Medical Science and Technology of Zhejiang Province(Nos.2019ZH022 and 2018KY478)the Zhejiang Province Public Welfare Technology Application Research Project(No.LGF19H180021),China
文摘Common bile duct(CBD)stones are a frequent problem in Chinese populations,and their incidence is particularly high in certain areas(Wang et al.,2013).In recent years,laparoscopic common bile duct exploration(LCBDE)and endoscopic retrograde cholangiopancreatography(ERCP)have been the main surgical procedures for CBD stones,although each has different advantages and disadvantages in the treatment of choledocholithiasis(Loor et al.,2017;Zhou et al.,2017).
基金Project supported by the National Natural Science Foundation of China(No.81272671)the Foundation of Health and Family Planning Commission of Zhejiang Province(No.2015KYB218)China
文摘Objective： The aim of our study is to observe the impact of cholangiocarcinoma-derived exosomes on the antitumor activities of cytokine-induced killer （CIK） cells and then demonstrate the appropriate mechanism. Methods： Tumor-derived exosomes （TEXs）, which are derived from RBE cells （human cholangiocarcinoma line）, were collected by ultracentrifugation. CIK cells induced from peripheral blood were stimulated by TEXs. Fluorescence-activated cell sorting （FACS） was performed to determine the phenotypes of TEX-CIK and N-CIK （normal CIK） cells. The concen- trations of tumor necrosis factor-a （TNF-a） and perforin in the culture medium supematant were examined by using an enzyme-linked immunosorbent assay （ELISA） kit. A CCK-8 kit was used to evaluate the cytotoxic activity of the CIK cells to the RBE cell line. Results： The concentrations of TNF-a and perforin of the group TEX-CIK were 138.61 pg/ml and 2.41 ng/ml, respectively, lower than those of the group N-CIK 194.08 pg/ml （P〈0.01） and 3.39 ng/ml （P〈0.05）. The killing rate of the group TEX-CIK was 33.35%, lower than that of the group N-CIK （47,35% （P〈0.01））. The population of CD3＋, CD8＋, NK （CD56＋）, and CD3＋CD56＋ cells decreased in the TEX-CIK group （63.2±6.8）%, （2.5±1.0）%, （0.53±0.49）%, （0.45±0.42）%） compared with the N-CIK group （（90.3±7.3）%, （65.7±3.3）%, （4.2±1.2）%, （15.2±2.7）%）, P〈0.01. Conclusions： Our results suggest that RBE cells-derived exosomes inhibit the antitumor activity of CIK cells by down-regulating the population of CD3＋, CD8＋, NK （CD56＋）, and CD3＋CD56＋ cells and the secretion of TNF-a and perforin. TEX may play an important role in cholangiocarcinoma immune escape.
基金Project supported by the National Natural Science Foundation of China(No.81272671)the Health Bureau of Zhejiang Province(No.WKJ2013-2-018),China
文摘Background： Tumor-derived exosomes were considered to be potential candidates for tumor vaccines because they are abundant in immune-regulating proteins, whereas tumor exosomal miRNAs may induce immune tolerance, thereby having an opposite immune function. Objective： This study was designed to separate exosomal protein and depleted exosomal microRNAs （miRNAs）, increasing the immune activity of exosomes for activating dendritic cell/cytokine-induced killer cells （DC/CIKs） against pancreatic cancer （PC）. Methods： PC-derived exosomes （PEs） were extracted from cultured PANC-1 cell supernatants and then ruptured; this was followed by ultrafiltered exosome lysates （UELs）. DCs were stimulated with lipopolysaccharide （LPS）, PE, and UEL, followed by co-culture with CIKs. The anti-tumor effects of DC/CIKs against PC were evaluated by proliferation and killing rates, tumor ne- crosis factor-a （TNF-a） and perforin secretion. Exosomal miRNAs were depleted after lysis and ultrafiltration, while 128 proteins were retained, including several immune-activating proteins. Results： UEL-stimulated DC/CIKs showed a higher killing rate than LPS- and PE-stimulated DC/CIKs. Conclusions： miRNA-depleted exosome proteins may be promising agonists for specifically activating DC/CIKs against PC.
