目的:旨在制备负载Der f 1/IGF-1的聚乳酸-乙醇酸共聚物(poly lactide-co-glycolide,PLGA)纳米颗粒(Der f 1/IGF-1 NPs),探讨其促进Treg细胞生成的作用。方法:通过复乳法制备包裹Der f 1/IGF-1的NPs并分析其理化性质及体外累计释放率;...目的:旨在制备负载Der f 1/IGF-1的聚乳酸-乙醇酸共聚物(poly lactide-co-glycolide,PLGA)纳米颗粒(Der f 1/IGF-1 NPs),探讨其促进Treg细胞生成的作用。方法:通过复乳法制备包裹Der f 1/IGF-1的NPs并分析其理化性质及体外累计释放率;预处理BMDC将细胞分为Saline组、Blank NPs组、Der f 1/IGF-1组以及Der f 1/IGF-1 NPs组;利用ELISA实验测定IL-10及TGF-β的表达;采用流式细胞术检测Treg细胞的数量变化。结果:Der f 1/IGF-1 NPs为球形结构,具有良好的分散性,粒径均<200 nm,Zeta电位为负电荷且具有稳定的缓释效果;BMDC预处理后,相比较Blank NPs组,Der f 1/IGF-1 NPs组,BMDC细胞TGF-β、IL-10的表达水平明显增高,差异有统计学意义(P<0.001);且与CD4^(+)T细胞共培养后,Der f 1/IGF-1 NPs组Treg细胞生成的比例明显提高,差异有统计学意义(P<0.001)。结论:Der f 1/IGF-1 NPs在体外实验中可诱导Treg细胞生成。本研究为重建免疫耐受功能失常提供了一种新的、更有效的方法。展开更多
目的探讨自体DC-CIK细胞治疗晚期肺癌患者前后T淋巴细胞亚群表面分子表达情况。方法抽取63例肺癌晚期患者的外周血分离获得外周血单个核细胞(PBMC),将与患者病理分类相同的肺癌细胞株经反复冻融处理获得可溶性抗原,制备肿瘤相关抗原肽负...目的探讨自体DC-CIK细胞治疗晚期肺癌患者前后T淋巴细胞亚群表面分子表达情况。方法抽取63例肺癌晚期患者的外周血分离获得外周血单个核细胞(PBMC),将与患者病理分类相同的肺癌细胞株经反复冻融处理获得可溶性抗原,制备肿瘤相关抗原肽负载DC-CIK细胞,并进行质量控制后回输。应用流式细胞术检测DC-CIK细胞治疗晚期肺癌患者前后T淋巴细胞表面CD3^+HLA-DR^+、CD3^+CD25^+、CD3^+CD4^+CD25^+、CD3^+CD8^+HLA-DR^+、CD3^+CD8^+CD38^+细胞百分比及对比治疗前后表达情况。结果成功获得成熟DC细胞,流式细胞术检测证实DC细胞成熟的标志分子CD11c、CD83、CD86及HLA-DR的表达均较高,分别为51.6%±10.3%、50.8%±9.7%、48.9%±11.4%、61.2%±6.5%;成功获得CIK细胞细胞,流式细胞术结果显示,CD3^+CD56^+双阳性细胞比例占21.3%±7.6%。与治疗前相比,DC-CIK细胞治疗后CD3^+HLA-DR^+(12.71±1.54 vs 11.44±4.13,P<0.05)、CD3^+CD8^+HLA-DR^+(7.48±1.01 vs 5.20±1.01,P<0.01)、CD3^+CD8^+CD38^+(8.27±1.71 vs 6.47±1.99,P<0.01)表达明显升高,CD3^+CD4^+CD25^+(5.38±1.47 vs 6.12±0.67,P<0.05)表达明显降低。结论采用自体DC-CIK免疫细胞治疗晚期肺癌可以明显提高患者免疫力。展开更多
<strong>Objective:</strong> To analyze various immune cytokines (NKG2D, IL-12, IL-15, IL-18, DC cells, TNF-a, IFN-r) and peripheral blood of patients with non-small cell lung cancer (NSCLC) at different ti...<strong>Objective:</strong> To analyze various immune cytokines (NKG2D, IL-12, IL-15, IL-18, DC cells, TNF-a, IFN-r) and peripheral blood of patients with non-small cell lung cancer (NSCLC) at different times after chemotherapy. Changes in CD4+, CD8+, Th17 and IgG, IgM, and IgA levels. <strong>Methods:</strong> A total of 118 NSCLC patients who attended the Oncology Department of the Affiliated Hospital of Chengde Medical College from September 2018 to September 2021 were selected as the research objects, and the patients were analyzed at different time points (before chemotherapy, after the first chemotherapy, and after the second chemotherapy). The effects of NKG2D, IL-12, IL-15, IL-18, DC cells, TNF-A, IFN-r, CD4+, CD8+ Th17, IgG, IgM and IgA levels in peripheral blood at different time points (before chemotherapy, after the first chemotherapy and after the second chemotherapy) were analyzed. The changes of NKG2D, IL-12, IL-15, IL-18, DC cells, TNF-A, IFN-r and the levels of CD4+, CD8+ Th17, IgG, IgM and IgA in peripheral blood were compared at each time point. <strong>Results:</strong> NKG2D, IL-12, IL-15, IL-18, TNF-a, IFN-r gradually decreased before chemotherapy, one week after chemotherapy, and two weeks after chemotherapy, the difference was statistically significant, but DC cells were not significant Variety. CD4+ and CD8+ both increased significantly, and the levels of Th17, IgG, IgM, and IgA gradually decreased. <strong>Conclusion:</strong> In the course of chemotherapy, all immune factors except DC cells were significantly decreased compared with those before chemotherapy, and the decrease of immune factors except DC cells was positively correlated with the length of chemotherapy cycle. If additional immunotherapy is needed, it should be carried out in the early stage of chemotherapy.展开更多
<strong>Objective:</strong> To analyze the effects of chemotherapy on peripheral blood DC cells and related immune cytokines (NKG2D, DC cells, TNF-a, IFN-r, HMGB-1) in patients with non-small cell lung can...<strong>Objective:</strong> To analyze the effects of chemotherapy on peripheral blood DC cells and related immune cytokines (NKG2D, DC cells, TNF-a, IFN-r, HMGB-1) in patients with non-small cell lung cancer (NSCLC). <strong>Methods:</strong> Ninety-five NSCLC patients who attended the Oncology Department of the Affiliated Hospital of Chengde Medical College from September 2018 to February 2021 were selected as the research objects, and the changes in the expression levels of DC cells, NKG2D, TNF-a, IFN-r, HMGB-1 in the peripheral blood of patients at different time points (before chemotherapy, after the first chemotherapy, and after the second chemotherapy) were analyzed, and the correlation between DC cells in blood and NKG2D, TNF-a, IFN-r, HMGB-1 at each time point was explored. <strong>Results:</strong> The expression levels of NKG2D, TNF-a, IFN-r, and HMGB-1 in the peripheral blood of the patient before chemotherapy, after the first chemotherapy, and after the second chemotherapy gradually decreased, and there was no significant change in DC cells, except for DC cells at different times. The difference between each factor of each point was statistically significant (all P < 0.05). Pearson correlation analysis showed that there was no correlation between peripheral blood DC cells of patients at different time points and other factors. <strong>Conclusion:</strong> The decrease of other immune cytokines except DC cells in peripheral blood of patients with NSCLC after chemotherapy may be one of the mechanisms by which the patient’s immune function is suppressed. There is no correlation between DC cells and other factors.展开更多
Objective： To investigate the antitumor effect of endostatin combined with tumor antigen-pulsed dendritic cell （DC）-T cell therapy on lung cancer. Methods： Transplanted Lewis lung cancer （LLC） models of C57BL/6 ...Objective： To investigate the antitumor effect of endostatin combined with tumor antigen-pulsed dendritic cell （DC）-T cell therapy on lung cancer. Methods： Transplanted Lewis lung cancer （LLC） models of C57BL/6 mice were established by subcutaneous injection of LLC cells in left extremity axillary. Tumor antigen-pulsed DC-T cells from spleen cells and bone of mice were cultured in vitro. Tumor-bearing mice were randomly divided into three groups, including DC- T＋endostatin group, DC-T group, and phosphate-buffered saline （PBS） control group. Microvessel density （MVD） of tumor tissue in tumor-bearing mice was determined by immunohistochemistry （IHC）. The expressions of vascular endothelial growth factor （VEGF） and hypoxia-inducible factor-1α （HIF-1α） were determined by Western blotting and IHC staining. The proportions of CD8＋ T cells, mature dendritic cells （mDC）, tumor-associated macrophages [TAM （M1/M2）], and myeloid-derived suppressor cells （MDSC） in suspended cells of tumor tissue were determined by flow cytometry. The expressions of inter｜eukin （IL）-6, IL-10, IL-17, transforming growth factor-β（TGF-β） and interferon-γ （IFN-γ） in suspended cells of tumor tissue were detected by enzyme-linked immune sorbent assay （ELISA）. Results： DC-T cells combined with endostatin remarkably suppressed tumor growth. MVD of mice in DC- T＋endostatin group was significantly lower than that of the control group and DC-T monotherapy group. The expressions of VEGF, IL-6 and IL-17 in tumors were markedly decreased, but IFN-γ, and HIF-1α increased after treating with DC-T cells combined with endostatin, compared to control group and DC-T group. In the DC- T＋endostatin group, the proportions of MDSC and TAM （M2 type） were significantly decreased, mDC and TAM （Nil type） were up-regulated, and CD8＋ T cells were recruited to infiltrate tumors, in contrast to PBS control and DC-T monotherapy. DC-T cells combined with endostatin potently reduced the 展开更多
<Abstract>Background and Objective:Cytokine-induced killer(CIK) cells and autologous dendritic cells-CIK(DCCIK) cells co-cultured with autologous dendritic cells(DCs) and CIK cells are commonly used for immunoth...<Abstract>Background and Objective:Cytokine-induced killer(CIK) cells and autologous dendritic cells-CIK(DCCIK) cells co-cultured with autologous dendritic cells(DCs) and CIK cells are commonly used for immunotherapy recently.We compared the anti-tumor immune response of CIK cells,autologous DC-CIK cells,and semi-allogeneic DC-CIK cells to explore a more effective anti-tumor adoptive immunotherapy approach.Met hods:Peripheral monocytes were isolated from patients with renal carcinoma,lung cancer,or maxillary squamous cell carcinoma and their healthy adult children.Isolated cells were cultured and induced as DCs and CIK cells in vitro.CIK cells from patients were co-cultured with autologous DCs and DCs from their children respectively,generating DC-CIK cells and semi-allogeneic DC-CIK cells.The anti-tumor activities of autologous CIK cells,autologous DC-CIK cells,and semi-allogeneic DC-CIK cells were measured by LDH assay.Intracellular staining was used to test the secretion of cytokines.Flow cytometry was applied for detecting the phonotype changes of these three types of cells.Cell proliferation and cell apoptosis were detected by 5,6carboxyfluorescein diacetate succinimidyl ester(CFSE) and Annexin V/PI respectively.Result s:Compared with autologous CIK cells and DC-CIK cells,semi-allogeneic DC-CIK cells significantly enhanced the anti-tumor activity and IFN-γ secretion,reduced IL-4 secretion,increased the ratio of CD3+CD56+ cells and CD3+CD8+ cells,decreased the number of CD4+CD25+ cells,promoted cell proliferation,and lessened cell apoptosis.Conclusions:Semi-allogeneic DC-CIK cells had a stronger anti-tumor effect than did autologous CIK cells and DC-CIK cells.Our results provided experimental evidence for clinical application of DC-CIK cells.展开更多
文摘目的:旨在制备负载Der f 1/IGF-1的聚乳酸-乙醇酸共聚物(poly lactide-co-glycolide,PLGA)纳米颗粒(Der f 1/IGF-1 NPs),探讨其促进Treg细胞生成的作用。方法:通过复乳法制备包裹Der f 1/IGF-1的NPs并分析其理化性质及体外累计释放率;预处理BMDC将细胞分为Saline组、Blank NPs组、Der f 1/IGF-1组以及Der f 1/IGF-1 NPs组;利用ELISA实验测定IL-10及TGF-β的表达;采用流式细胞术检测Treg细胞的数量变化。结果:Der f 1/IGF-1 NPs为球形结构,具有良好的分散性,粒径均<200 nm,Zeta电位为负电荷且具有稳定的缓释效果;BMDC预处理后,相比较Blank NPs组,Der f 1/IGF-1 NPs组,BMDC细胞TGF-β、IL-10的表达水平明显增高,差异有统计学意义(P<0.001);且与CD4^(+)T细胞共培养后,Der f 1/IGF-1 NPs组Treg细胞生成的比例明显提高,差异有统计学意义(P<0.001)。结论:Der f 1/IGF-1 NPs在体外实验中可诱导Treg细胞生成。本研究为重建免疫耐受功能失常提供了一种新的、更有效的方法。
文摘目的探讨自体DC-CIK细胞治疗晚期肺癌患者前后T淋巴细胞亚群表面分子表达情况。方法抽取63例肺癌晚期患者的外周血分离获得外周血单个核细胞(PBMC),将与患者病理分类相同的肺癌细胞株经反复冻融处理获得可溶性抗原,制备肿瘤相关抗原肽负载DC-CIK细胞,并进行质量控制后回输。应用流式细胞术检测DC-CIK细胞治疗晚期肺癌患者前后T淋巴细胞表面CD3^+HLA-DR^+、CD3^+CD25^+、CD3^+CD4^+CD25^+、CD3^+CD8^+HLA-DR^+、CD3^+CD8^+CD38^+细胞百分比及对比治疗前后表达情况。结果成功获得成熟DC细胞,流式细胞术检测证实DC细胞成熟的标志分子CD11c、CD83、CD86及HLA-DR的表达均较高,分别为51.6%±10.3%、50.8%±9.7%、48.9%±11.4%、61.2%±6.5%;成功获得CIK细胞细胞,流式细胞术结果显示,CD3^+CD56^+双阳性细胞比例占21.3%±7.6%。与治疗前相比,DC-CIK细胞治疗后CD3^+HLA-DR^+(12.71±1.54 vs 11.44±4.13,P<0.05)、CD3^+CD8^+HLA-DR^+(7.48±1.01 vs 5.20±1.01,P<0.01)、CD3^+CD8^+CD38^+(8.27±1.71 vs 6.47±1.99,P<0.01)表达明显升高,CD3^+CD4^+CD25^+(5.38±1.47 vs 6.12±0.67,P<0.05)表达明显降低。结论采用自体DC-CIK免疫细胞治疗晚期肺癌可以明显提高患者免疫力。
文摘<strong>Objective:</strong> To analyze various immune cytokines (NKG2D, IL-12, IL-15, IL-18, DC cells, TNF-a, IFN-r) and peripheral blood of patients with non-small cell lung cancer (NSCLC) at different times after chemotherapy. Changes in CD4+, CD8+, Th17 and IgG, IgM, and IgA levels. <strong>Methods:</strong> A total of 118 NSCLC patients who attended the Oncology Department of the Affiliated Hospital of Chengde Medical College from September 2018 to September 2021 were selected as the research objects, and the patients were analyzed at different time points (before chemotherapy, after the first chemotherapy, and after the second chemotherapy). The effects of NKG2D, IL-12, IL-15, IL-18, DC cells, TNF-A, IFN-r, CD4+, CD8+ Th17, IgG, IgM and IgA levels in peripheral blood at different time points (before chemotherapy, after the first chemotherapy and after the second chemotherapy) were analyzed. The changes of NKG2D, IL-12, IL-15, IL-18, DC cells, TNF-A, IFN-r and the levels of CD4+, CD8+ Th17, IgG, IgM and IgA in peripheral blood were compared at each time point. <strong>Results:</strong> NKG2D, IL-12, IL-15, IL-18, TNF-a, IFN-r gradually decreased before chemotherapy, one week after chemotherapy, and two weeks after chemotherapy, the difference was statistically significant, but DC cells were not significant Variety. CD4+ and CD8+ both increased significantly, and the levels of Th17, IgG, IgM, and IgA gradually decreased. <strong>Conclusion:</strong> In the course of chemotherapy, all immune factors except DC cells were significantly decreased compared with those before chemotherapy, and the decrease of immune factors except DC cells was positively correlated with the length of chemotherapy cycle. If additional immunotherapy is needed, it should be carried out in the early stage of chemotherapy.
文摘<strong>Objective:</strong> To analyze the effects of chemotherapy on peripheral blood DC cells and related immune cytokines (NKG2D, DC cells, TNF-a, IFN-r, HMGB-1) in patients with non-small cell lung cancer (NSCLC). <strong>Methods:</strong> Ninety-five NSCLC patients who attended the Oncology Department of the Affiliated Hospital of Chengde Medical College from September 2018 to February 2021 were selected as the research objects, and the changes in the expression levels of DC cells, NKG2D, TNF-a, IFN-r, HMGB-1 in the peripheral blood of patients at different time points (before chemotherapy, after the first chemotherapy, and after the second chemotherapy) were analyzed, and the correlation between DC cells in blood and NKG2D, TNF-a, IFN-r, HMGB-1 at each time point was explored. <strong>Results:</strong> The expression levels of NKG2D, TNF-a, IFN-r, and HMGB-1 in the peripheral blood of the patient before chemotherapy, after the first chemotherapy, and after the second chemotherapy gradually decreased, and there was no significant change in DC cells, except for DC cells at different times. The difference between each factor of each point was statistically significant (all P < 0.05). Pearson correlation analysis showed that there was no correlation between peripheral blood DC cells of patients at different time points and other factors. <strong>Conclusion:</strong> The decrease of other immune cytokines except DC cells in peripheral blood of patients with NSCLC after chemotherapy may be one of the mechanisms by which the patient’s immune function is suppressed. There is no correlation between DC cells and other factors.
基金supported by Natural Science Foundation of Shandong province,China(No.ZR2010HL015)Natural Science Youth Foundation of Shandong province,China(No.ZR2013HQ017)
文摘Objective： To investigate the antitumor effect of endostatin combined with tumor antigen-pulsed dendritic cell （DC）-T cell therapy on lung cancer. Methods： Transplanted Lewis lung cancer （LLC） models of C57BL/6 mice were established by subcutaneous injection of LLC cells in left extremity axillary. Tumor antigen-pulsed DC-T cells from spleen cells and bone of mice were cultured in vitro. Tumor-bearing mice were randomly divided into three groups, including DC- T＋endostatin group, DC-T group, and phosphate-buffered saline （PBS） control group. Microvessel density （MVD） of tumor tissue in tumor-bearing mice was determined by immunohistochemistry （IHC）. The expressions of vascular endothelial growth factor （VEGF） and hypoxia-inducible factor-1α （HIF-1α） were determined by Western blotting and IHC staining. The proportions of CD8＋ T cells, mature dendritic cells （mDC）, tumor-associated macrophages [TAM （M1/M2）], and myeloid-derived suppressor cells （MDSC） in suspended cells of tumor tissue were determined by flow cytometry. The expressions of inter｜eukin （IL）-6, IL-10, IL-17, transforming growth factor-β（TGF-β） and interferon-γ （IFN-γ） in suspended cells of tumor tissue were detected by enzyme-linked immune sorbent assay （ELISA）. Results： DC-T cells combined with endostatin remarkably suppressed tumor growth. MVD of mice in DC- T＋endostatin group was significantly lower than that of the control group and DC-T monotherapy group. The expressions of VEGF, IL-6 and IL-17 in tumors were markedly decreased, but IFN-γ, and HIF-1α increased after treating with DC-T cells combined with endostatin, compared to control group and DC-T group. In the DC- T＋endostatin group, the proportions of MDSC and TAM （M2 type） were significantly decreased, mDC and TAM （Nil type） were up-regulated, and CD8＋ T cells were recruited to infiltrate tumors, in contrast to PBS control and DC-T monotherapy. DC-T cells combined with endostatin potently reduced the
基金supported by Poten Biomedical Technology Development Co.,Ltd.
文摘<Abstract>Background and Objective:Cytokine-induced killer(CIK) cells and autologous dendritic cells-CIK(DCCIK) cells co-cultured with autologous dendritic cells(DCs) and CIK cells are commonly used for immunotherapy recently.We compared the anti-tumor immune response of CIK cells,autologous DC-CIK cells,and semi-allogeneic DC-CIK cells to explore a more effective anti-tumor adoptive immunotherapy approach.Met hods:Peripheral monocytes were isolated from patients with renal carcinoma,lung cancer,or maxillary squamous cell carcinoma and their healthy adult children.Isolated cells were cultured and induced as DCs and CIK cells in vitro.CIK cells from patients were co-cultured with autologous DCs and DCs from their children respectively,generating DC-CIK cells and semi-allogeneic DC-CIK cells.The anti-tumor activities of autologous CIK cells,autologous DC-CIK cells,and semi-allogeneic DC-CIK cells were measured by LDH assay.Intracellular staining was used to test the secretion of cytokines.Flow cytometry was applied for detecting the phonotype changes of these three types of cells.Cell proliferation and cell apoptosis were detected by 5,6carboxyfluorescein diacetate succinimidyl ester(CFSE) and Annexin V/PI respectively.Result s:Compared with autologous CIK cells and DC-CIK cells,semi-allogeneic DC-CIK cells significantly enhanced the anti-tumor activity and IFN-γ secretion,reduced IL-4 secretion,increased the ratio of CD3+CD56+ cells and CD3+CD8+ cells,decreased the number of CD4+CD25+ cells,promoted cell proliferation,and lessened cell apoptosis.Conclusions:Semi-allogeneic DC-CIK cells had a stronger anti-tumor effect than did autologous CIK cells and DC-CIK cells.Our results provided experimental evidence for clinical application of DC-CIK cells.
