AIM: Cytotoxic T lymphocytes (CTLs) play an important role in resolving HBV infection. In the present study, we attempted to evaluate the efficiency of bone marrow-derived dendritic cells (DCs) transduced with recombi...AIM: Cytotoxic T lymphocytes (CTLs) play an important role in resolving HBV infection. In the present study, we attempted to evaluate the efficiency of bone marrow-derived dendritic cells (DCs) transduced with recombinant retroviral vector bearing hepatitis B virus (HBV) core gene and the capability of generating CTLs against HBcAg by genetically modified DCs in vivo.METHODS: A retroviral vector containing HBV core gene was constructed. Replicating DC progenitor of C57BL/6 mice was transduced by retroviral vector and continually cultured in the presence of recombinant mouse granulocytemacrophage colony-stimulating factor (rmGM-CSF) and interleukin-4(IL-4) for 6 days. LPS was added and cultured for additional two days. The efficiency of gene transfer was determined by PCR, Western blot and FACS. Transduced DCs immunized C57BL/6 mice subcutaneously 2 times at an oneweek interval. Intracellular IFN-γ and IL-4 of immunized mice lymphocytes were analyzed. Generation of CTLs in lymphooytes stimulated with mitomycin C-treated EL4-C cell which stably expresses HBcAg was determined by LDH release assays.RESULTS: Recombinant retroviral expression vector (pLCSN) was positively detected by PCR as well as enzyme digestion with EcoRI and BamH I. Retroviruses were generated by pLCSN transfection packing cell and the virus titer was 3x10s CFU/ml. Indirect immunofluorescence and FACS showed that HBV core gene was expressed in murine fibroblasts. Transduced bone marrow cells had capability of differentiating into DCsc in vitro in the presence of rmGMCSF and rmIL-4. The result of PCR showed that HBV core gene was integrated into the genome of transduced DCs.Western blot analysis showed that HBV core gene was expressed in DCs. The transduction rate was 28 % determined by FACS. Retroviral transduction had no influence on DCs expressions of CD80 and MHC class Ⅱ. HBcAg specific CTLs and Th1 type immune responses could be generated in the mice by using transduced DCs as antigen presenting cells (APCs).CONCLUSION: Retroviral transduct展开更多
Immunology-based therapy is rapidly developing into an effective treatment option for a surprising range of cancers. We have learned over the last decade that powerful immunologic effector cells may be blocked by inhi...Immunology-based therapy is rapidly developing into an effective treatment option for a surprising range of cancers. We have learned over the last decade that powerful immunologic effector cells may be blocked by inhibitory regulatory pathways controlled by specific molecules often called "immune checkpoints." These checkpoints serve to control or turn off the immune response when it is no longer needed to prevent tissue injury and autoimmunity. Cancer cells have learned or evolved to use these mechanisms to evade immune control and elimination. The development of a new therapeutic class of drugs that inhibit these inhibitory pathways has recently emerged as a potent strategy in oncology. Three sets of agents have emerged in clinical trials exploiting this strategy. These agents are antibodybased therapies targeting cytotoxic T-lymphocyte antigen 4(CTLA4), programmed cell death 1(PD-1), and programmed cell death ligand 1(PD-L1). These inhibitors of immune inhibition have demonstrated extensive activity as single agents and in combinations. Clinical responses have been seen in melanoma, renal cell carcinoma, non-small cell lung cancer, and several other tumor types. Despite the autoimmune or inflammatory immune-mediated adverse effects which have been seen, the responses and overall survival benefits exhibited thus far warrant further clinical development.展开更多
OBJECTIVE： Few studies have investigated the ef- fects produced by combinations of polysaccharides and chemotherapeutic drugs in cancer treatment. We hypothesized that a combination of polysaccha- rides （COP） from ...OBJECTIVE： Few studies have investigated the ef- fects produced by combinations of polysaccharides and chemotherapeutic drugs in cancer treatment. We hypothesized that a combination of polysaccha- rides （COP） from Lentinus edodes and Tricholoma matsutake would improve the efficacy of 5-fluoro- uracil （5-FU）-mediated inhibition of H22 cell growth. METHODS： Mice were injected H22 cells and then treated with either 5-FU, polysaccharides from Tri- choloma matsutake （PTM）, polysaccharides from Lentinus edodes （PL）, PTM ＋ PL, 5-FU ＋ PTM, 5-FU ＋ PL, or 5-FU ＋ COR The tumor weight and volume, and splenic CD4 ＋ and CD8 ＋ T cell frequencies, were determined. Additionally, splenic natural killer （NK） cell and cytotoxic T lymphocyte （CTL） activities were assessed and the serum levels of tumor necro- sis factor-a （TNF-a）, Interleukin-2 （IL-2）, and Interfer- on-y （IFN-y） were measured. RESULTS： Compared with mice from the control,5-FU, PL, PTM, PTM ＋ PL, 5-FU ＋ PL, and 5-FU ＋ PTM groups, mice treated with 5-FU ＋ COP showed： （a） significantly reduced tumor weight and volume （P〈 0.05）, （b） significantly higher serum levels of TNF-α, IL-2, and IFN-y （P〈0.05）, （c） significantly increased CD4＋ and CD8＋ 1 cell frequencies in the spleen （P〈 0.05）, and （d） significantly increased splenic NK cell and CTL activities （P〈0.05）. The tumor weight and volume in mice treated with 5-FU＋PL or 5-FU＋PTM were significantly reduced compared with mice treated with 5-FU alone （P〈0.05）. Serum levels of TNF-α, 11.-2, and IFN-y, frequencies of CD4 ＋ and CD8＋ T cells in the spleen, and splenic NK and CTL activities were also significantly increased in mice treated with 5-FU＋PL or 5-FU＋PTM compared with mice treated with 5-FU alone （P〈0.05）. CONCLUSION： Polysaccharides from Lentinus edodes and Tricholoma matsutake could enhance the efficacy of 5-FU-mediated H22 cell growth inhi- bition.展开更多
机体的免疫系统可以识别并摧毁肿瘤,但肿瘤为了逃避免疫攻击可以进化。当前以细胞毒性T淋巴细胞相关蛋白4(cytotoxic T-lymphocyte-associated protein 4,CTLA4)或程序性死亡分子受体1(programmed death 1,PD-1)及程序性死亡分子配体1(p...机体的免疫系统可以识别并摧毁肿瘤,但肿瘤为了逃避免疫攻击可以进化。当前以细胞毒性T淋巴细胞相关蛋白4(cytotoxic T-lymphocyte-associated protein 4,CTLA4)或程序性死亡分子受体1(programmed death 1,PD-1)及程序性死亡分子配体1(programmed death 1 ligand,PD-L1)为靶点的免疫治疗成为了强有力的新治疗方案[1]。展开更多
Unprecedented progress has seen made in the last decade in the fi eld of cancer immunotherapy.The recent approval of nivolumab(Opdivo),the fi rst anti-programmed cell death-1(PD-1)antibody,for metastatic melanoma in J...Unprecedented progress has seen made in the last decade in the fi eld of cancer immunotherapy.The recent approval of nivolumab(Opdivo),the fi rst anti-programmed cell death-1(PD-1)antibody,for metastatic melanoma in Japan,marked a milestone in the rapidly advancing fi eld of cancer immunotherapy.Nivolumab together展开更多
基金a grant from Key Lab Programs of Jiangsu Province,No.k2030
文摘AIM: Cytotoxic T lymphocytes (CTLs) play an important role in resolving HBV infection. In the present study, we attempted to evaluate the efficiency of bone marrow-derived dendritic cells (DCs) transduced with recombinant retroviral vector bearing hepatitis B virus (HBV) core gene and the capability of generating CTLs against HBcAg by genetically modified DCs in vivo.METHODS: A retroviral vector containing HBV core gene was constructed. Replicating DC progenitor of C57BL/6 mice was transduced by retroviral vector and continually cultured in the presence of recombinant mouse granulocytemacrophage colony-stimulating factor (rmGM-CSF) and interleukin-4(IL-4) for 6 days. LPS was added and cultured for additional two days. The efficiency of gene transfer was determined by PCR, Western blot and FACS. Transduced DCs immunized C57BL/6 mice subcutaneously 2 times at an oneweek interval. Intracellular IFN-γ and IL-4 of immunized mice lymphocytes were analyzed. Generation of CTLs in lymphooytes stimulated with mitomycin C-treated EL4-C cell which stably expresses HBcAg was determined by LDH release assays.RESULTS: Recombinant retroviral expression vector (pLCSN) was positively detected by PCR as well as enzyme digestion with EcoRI and BamH I. Retroviruses were generated by pLCSN transfection packing cell and the virus titer was 3x10s CFU/ml. Indirect immunofluorescence and FACS showed that HBV core gene was expressed in murine fibroblasts. Transduced bone marrow cells had capability of differentiating into DCsc in vitro in the presence of rmGMCSF and rmIL-4. The result of PCR showed that HBV core gene was integrated into the genome of transduced DCs.Western blot analysis showed that HBV core gene was expressed in DCs. The transduction rate was 28 % determined by FACS. Retroviral transduction had no influence on DCs expressions of CD80 and MHC class Ⅱ. HBcAg specific CTLs and Th1 type immune responses could be generated in the mice by using transduced DCs as antigen presenting cells (APCs).CONCLUSION: Retroviral transduct
文摘Immunology-based therapy is rapidly developing into an effective treatment option for a surprising range of cancers. We have learned over the last decade that powerful immunologic effector cells may be blocked by inhibitory regulatory pathways controlled by specific molecules often called "immune checkpoints." These checkpoints serve to control or turn off the immune response when it is no longer needed to prevent tissue injury and autoimmunity. Cancer cells have learned or evolved to use these mechanisms to evade immune control and elimination. The development of a new therapeutic class of drugs that inhibit these inhibitory pathways has recently emerged as a potent strategy in oncology. Three sets of agents have emerged in clinical trials exploiting this strategy. These agents are antibodybased therapies targeting cytotoxic T-lymphocyte antigen 4(CTLA4), programmed cell death 1(PD-1), and programmed cell death ligand 1(PD-L1). These inhibitors of immune inhibition have demonstrated extensive activity as single agents and in combinations. Clinical responses have been seen in melanoma, renal cell carcinoma, non-small cell lung cancer, and several other tumor types. Despite the autoimmune or inflammatory immune-mediated adverse effects which have been seen, the responses and overall survival benefits exhibited thus far warrant further clinical development.
基金Supported by Department of Science and Technology of Jilin Province(No.yyzx201123-2No.20130522056JH)
文摘OBJECTIVE： Few studies have investigated the ef- fects produced by combinations of polysaccharides and chemotherapeutic drugs in cancer treatment. We hypothesized that a combination of polysaccha- rides （COP） from Lentinus edodes and Tricholoma matsutake would improve the efficacy of 5-fluoro- uracil （5-FU）-mediated inhibition of H22 cell growth. METHODS： Mice were injected H22 cells and then treated with either 5-FU, polysaccharides from Tri- choloma matsutake （PTM）, polysaccharides from Lentinus edodes （PL）, PTM ＋ PL, 5-FU ＋ PTM, 5-FU ＋ PL, or 5-FU ＋ COR The tumor weight and volume, and splenic CD4 ＋ and CD8 ＋ T cell frequencies, were determined. Additionally, splenic natural killer （NK） cell and cytotoxic T lymphocyte （CTL） activities were assessed and the serum levels of tumor necro- sis factor-a （TNF-a）, Interleukin-2 （IL-2）, and Interfer- on-y （IFN-y） were measured. RESULTS： Compared with mice from the control,5-FU, PL, PTM, PTM ＋ PL, 5-FU ＋ PL, and 5-FU ＋ PTM groups, mice treated with 5-FU ＋ COP showed： （a） significantly reduced tumor weight and volume （P〈 0.05）, （b） significantly higher serum levels of TNF-α, IL-2, and IFN-y （P〈0.05）, （c） significantly increased CD4＋ and CD8＋ 1 cell frequencies in the spleen （P〈 0.05）, and （d） significantly increased splenic NK cell and CTL activities （P〈0.05）. The tumor weight and volume in mice treated with 5-FU＋PL or 5-FU＋PTM were significantly reduced compared with mice treated with 5-FU alone （P〈0.05）. Serum levels of TNF-α, 11.-2, and IFN-y, frequencies of CD4 ＋ and CD8＋ T cells in the spleen, and splenic NK and CTL activities were also significantly increased in mice treated with 5-FU＋PL or 5-FU＋PTM compared with mice treated with 5-FU alone （P〈0.05）. CONCLUSION： Polysaccharides from Lentinus edodes and Tricholoma matsutake could enhance the efficacy of 5-FU-mediated H22 cell growth inhi- bition.
文摘机体的免疫系统可以识别并摧毁肿瘤,但肿瘤为了逃避免疫攻击可以进化。当前以细胞毒性T淋巴细胞相关蛋白4(cytotoxic T-lymphocyte-associated protein 4,CTLA4)或程序性死亡分子受体1(programmed death 1,PD-1)及程序性死亡分子配体1(programmed death 1 ligand,PD-L1)为靶点的免疫治疗成为了强有力的新治疗方案[1]。
文摘Unprecedented progress has seen made in the last decade in the fi eld of cancer immunotherapy.The recent approval of nivolumab(Opdivo),the fi rst anti-programmed cell death-1(PD-1)antibody,for metastatic melanoma in Japan,marked a milestone in the rapidly advancing fi eld of cancer immunotherapy.Nivolumab together
