Objective The aim of this study was to purify effective tumor peptide complexes from human melanoma cell lines to enhance the treatment effects on melanoma.Methods We purified heat shock protein 70(HSP70)-peptide comp...Objective The aim of this study was to purify effective tumor peptide complexes from human melanoma cell lines to enhance the treatment effects on melanoma.Methods We purified heat shock protein 70(HSP70)-peptide complexes(PCs)from human melanoma cell lines A375,A875,M21,M14,WM-35,and SK-HEL-1.We named the purified product as M-HSP70-PCs and determined its immunological activities.Autologous HSP70-PCs purified from primary tumor cells of melanoma patients(9 cases)were used as controls.These two tumor antigenic complexes were loaded into dendritic cells(DCs)and used to stimulate an antitumor response against tumor cells in the corresponding patients.Results Mature DCs pulsed with M-HSP70-PCs stimulated autologous T cells to secrete the same levels of type I cytokines as the autologous HSP70-PCs.Moreover,DCs pulsed with M-HSP70-PCs endued CIK cells with an equal ability as autologous HSP70-PCs to kill melanoma cells in the patients.Conclusion M-HSP70-PCs may be used as an efficient and generalized tumor antigen in the treatment of DC-based malignant melanoma.展开更多
Objective The successful establishment of a tumor cell bank is based on the premise that the target cells can be cultured by a legitimate approach.In this experiment,we used primary culture to select and detect breast...Objective The successful establishment of a tumor cell bank is based on the premise that the target cells can be cultured by a legitimate approach.In this experiment,we used primary culture to select and detect breast cancer cells in vitro,which can provide experimental ideas and methods for the establishment of a living tumor tissue cell bank.Methods Fifty-two specimens were collected over a two-year period from people with breast cancer who needed surgical treatment in our hospital.Cells were isolated and used to establish successful cell culture.Cell activity and cell purity were measured before liquid nitrogen cryopreservation.Results(1)At the initial culture stage,cells grew with adherence.Cell multiplication could be seen after the cell medium was exchanged three times.Cell viability was above 86%,while the viability of the target cells was above 75%,as detected by hematoxylin and eosin(HE)staining.(2)The number of breast cancer cells decreased,while the number of fibroblasts increased after five rounds of passage.(3)The success rate was 73.08%,which did not include polluted cells and those that were not successfully cryopreserved.Conclusion(1)breast cancer cells could be selected from primary culture in vitro through an appropriate method.(2)Exchange of the cell medium and further cell passage improved cell multiplication.(3)The experimental results could be monitored using trypan blue and HE staining.(4)The success of breast cancer cell culture in vitro could be used as a reference for other cell culture,so as to establish a tumor tissue cell bank.展开更多
Objective This study aimed to compare the anti-tumor effects of cytokine-induced killer (CIK) cellsinduced by autologous cytokines in patients with breast cancer and those of allogeneic CIK cells fromhealthy adults.Me...Objective This study aimed to compare the anti-tumor effects of cytokine-induced killer (CIK) cellsinduced by autologous cytokines in patients with breast cancer and those of allogeneic CIK cells fromhealthy adults.Methods We used conventional methods to induce CIK cells originating from two peripheral bloodmononuclear cell types (from patients with breast cancer and healthy adults). Killing activity was detectedusing an LDH assay, immunophenotypic changes were analyzed by flow cytometry, and the IFN-γ level ofculture supernatants was detected by ELISA.Results The results showed that the proliferative capacity of the allogeneic CIK cells was significantlyhigher than that of the autologous CIK cells. Compared with autologous CIK cells, the allogeneic CIK cellshad significantly enhanced anti-tumor activity against SKBR-3 cells (P < 0.01) and IFN-γ secretion (P <0.05);moreover, they increased the ratio of CD3+ CD56+ cells and CD3+ CD8+ cells (P < 0.05).Conclusion Healthy adult-derived induced CIK cells exhibited a stronger anti-tumor effect than inducedCIK cells derived from patients with breast cancer. The results of this study could provide experimentalevidence for the clinical application of CIK cells.展开更多
Objective The aim of this study was to enhance the treatment effect of tumor purified autogenous heat shock protein 70-peptide complexes(HSP70-PCs)on HER-3-overexpressing breast cancer.Methods In this study,we first s...Objective The aim of this study was to enhance the treatment effect of tumor purified autogenous heat shock protein 70-peptide complexes(HSP70-PCs)on HER-3-overexpressing breast cancer.Methods In this study,we first studied the expression of HER-3 in breast cancer tissues and its relationship with patient characteristics.We then purified HSP70-PCs from primary breast cancer cells with different HER-2 and HER-3 expression profiles and determined the cytotoxicity of autogenous dendritic cells(DCs)and CD8+T cells induced by these complexes.Third,recombinant human HSP70-HER-3 protein complexes were used to inhibit the autogenous HSP70-PCs purified from HER-3-overexpressing breast cancer cells,and the resulting immunological response was examined.Results The results show that HSP70-PCs can be combined with recombinant HSP70-HER-3 protein complexes to induce stronger immunological responses than autogenous HSP70-PCs alone and that these treatments induce autogenous CD8+T cell killing of HER-3-positive breast cancer cells.Conclusion These findings provide a new direction for HSP70-DC-based immunotherapy for patients with HER-3-overexpressing breast cancer.展开更多
基金Supported by a grant from the National Natural Sciences Foundation of Inner Mongolia(No.2017MS(LH)0847)。
文摘Objective The aim of this study was to purify effective tumor peptide complexes from human melanoma cell lines to enhance the treatment effects on melanoma.Methods We purified heat shock protein 70(HSP70)-peptide complexes(PCs)from human melanoma cell lines A375,A875,M21,M14,WM-35,and SK-HEL-1.We named the purified product as M-HSP70-PCs and determined its immunological activities.Autologous HSP70-PCs purified from primary tumor cells of melanoma patients(9 cases)were used as controls.These two tumor antigenic complexes were loaded into dendritic cells(DCs)and used to stimulate an antitumor response against tumor cells in the corresponding patients.Results Mature DCs pulsed with M-HSP70-PCs stimulated autologous T cells to secrete the same levels of type I cytokines as the autologous HSP70-PCs.Moreover,DCs pulsed with M-HSP70-PCs endued CIK cells with an equal ability as autologous HSP70-PCs to kill melanoma cells in the patients.Conclusion M-HSP70-PCs may be used as an efficient and generalized tumor antigen in the treatment of DC-based malignant melanoma.
基金a grant from the Science Foundation of Inner Mongolia Autonomous Region People’s Hospital(No.2016015).
文摘Objective The successful establishment of a tumor cell bank is based on the premise that the target cells can be cultured by a legitimate approach.In this experiment,we used primary culture to select and detect breast cancer cells in vitro,which can provide experimental ideas and methods for the establishment of a living tumor tissue cell bank.Methods Fifty-two specimens were collected over a two-year period from people with breast cancer who needed surgical treatment in our hospital.Cells were isolated and used to establish successful cell culture.Cell activity and cell purity were measured before liquid nitrogen cryopreservation.Results(1)At the initial culture stage,cells grew with adherence.Cell multiplication could be seen after the cell medium was exchanged three times.Cell viability was above 86%,while the viability of the target cells was above 75%,as detected by hematoxylin and eosin(HE)staining.(2)The number of breast cancer cells decreased,while the number of fibroblasts increased after five rounds of passage.(3)The success rate was 73.08%,which did not include polluted cells and those that were not successfully cryopreserved.Conclusion(1)breast cancer cells could be selected from primary culture in vitro through an appropriate method.(2)Exchange of the cell medium and further cell passage improved cell multiplication.(3)The experimental results could be monitored using trypan blue and HE staining.(4)The success of breast cancer cell culture in vitro could be used as a reference for other cell culture,so as to establish a tumor tissue cell bank.
基金Supported by a grant from the National Natural Sciences Foundation of Inner Mongolia(No.2012MS1102).
文摘Objective This study aimed to compare the anti-tumor effects of cytokine-induced killer (CIK) cellsinduced by autologous cytokines in patients with breast cancer and those of allogeneic CIK cells fromhealthy adults.Methods We used conventional methods to induce CIK cells originating from two peripheral bloodmononuclear cell types (from patients with breast cancer and healthy adults). Killing activity was detectedusing an LDH assay, immunophenotypic changes were analyzed by flow cytometry, and the IFN-γ level ofculture supernatants was detected by ELISA.Results The results showed that the proliferative capacity of the allogeneic CIK cells was significantlyhigher than that of the autologous CIK cells. Compared with autologous CIK cells, the allogeneic CIK cellshad significantly enhanced anti-tumor activity against SKBR-3 cells (P < 0.01) and IFN-γ secretion (P <0.05);moreover, they increased the ratio of CD3+ CD56+ cells and CD3+ CD8+ cells (P < 0.05).Conclusion Healthy adult-derived induced CIK cells exhibited a stronger anti-tumor effect than inducedCIK cells derived from patients with breast cancer. The results of this study could provide experimentalevidence for the clinical application of CIK cells.
基金Supported by a grant from the National Natural Science Foundation of China(No.81260392).
文摘Objective The aim of this study was to enhance the treatment effect of tumor purified autogenous heat shock protein 70-peptide complexes(HSP70-PCs)on HER-3-overexpressing breast cancer.Methods In this study,we first studied the expression of HER-3 in breast cancer tissues and its relationship with patient characteristics.We then purified HSP70-PCs from primary breast cancer cells with different HER-2 and HER-3 expression profiles and determined the cytotoxicity of autogenous dendritic cells(DCs)and CD8+T cells induced by these complexes.Third,recombinant human HSP70-HER-3 protein complexes were used to inhibit the autogenous HSP70-PCs purified from HER-3-overexpressing breast cancer cells,and the resulting immunological response was examined.Results The results show that HSP70-PCs can be combined with recombinant HSP70-HER-3 protein complexes to induce stronger immunological responses than autogenous HSP70-PCs alone and that these treatments induce autogenous CD8+T cell killing of HER-3-positive breast cancer cells.Conclusion These findings provide a new direction for HSP70-DC-based immunotherapy for patients with HER-3-overexpressing breast cancer.
