From the mouse hybridoma cell line secreting an anti-CD4 monoclonal antibody (McAb), total RNA was prepared. The VH and VL genes were amplified by RT-PCR with family specific primer pairs. The PCR products were cloned...From the mouse hybridoma cell line secreting an anti-CD4 monoclonal antibody (McAb), total RNA was prepared. The VH and VL genes were amplified by RT-PCR with family specific primer pairs. The PCR products were cloned into pGEM-T vectors, then tranfected into JM109. The VH and VL genes were snalyzed by automatic DNA sequencer. According to Kabat classification, the VH and VL genes belong to the mouse ig heavy subgroup Ⅱ(A) and x chain subgroupⅢ, respectively. The VH and VL genes were subcloned into pr1-Expr and Pk Expr respectively, then transfected into XL2-Blue. The VH- Pr1 and VL- pk were trans feeted by electroporation into mouse myeloma cell X63Ag8. 653. The transfectoma cells were selected by G418 screening, and then supernatant of cultured transfectoma were analyzed by ELISA and immunofluorescence techniques.We have acquired transfectoma cells secreting anti-CD4 chimeric antibodies.These chimeric antibodies are able to kill tumor cells specifically in vitro.展开更多
Adoptive cell therapy(ACT)is an emerging powerful cancer immunotherapy,which includes a complex process of genetic modification,stimulation and expansion.During these in vitro or ex vivo manipulation,sensitive cells a...Adoptive cell therapy(ACT)is an emerging powerful cancer immunotherapy,which includes a complex process of genetic modification,stimulation and expansion.During these in vitro or ex vivo manipulation,sensitive cells are inescapability subjected to harmful external stimuli.Although a variety of cytoprotection strategies have been developed,their application on ACT remains challenging.Herein,a DNA network is constructed on cell surface by rolling circle amplification(RCA),and T cell-targeted trivalent tetrahedral DNA nanostructure is used as a rigid scaffold to achieve high-efficient and selective coating for T cells.The cytoprotective DNA network on T-cell surface makes them aggregate over time to form cell clusters,which exhibit more resistance to external stimuli and enhanced activities in human peripheral blood mononuclear cells and liver cancer organoid killing model.Overall,this work provides a novel strategy for in vitro T cell-selective protection,which has a great potential for application in ACT.展开更多
目的考察Fe_(3)O_(4)纳米颗粒在低频振动磁场(low-frequency vibrating magnetic field,VMF)驱动下通过磁场机械力杀伤肿瘤细胞的效果。方法通过共沉淀法合成一种磁性强、具有不规则形貌的立方相Fe_(3)O_(4)纳米颗粒。将其置于本课题组...目的考察Fe_(3)O_(4)纳米颗粒在低频振动磁场(low-frequency vibrating magnetic field,VMF)驱动下通过磁场机械力杀伤肿瘤细胞的效果。方法通过共沉淀法合成一种磁性强、具有不规则形貌的立方相Fe_(3)O_(4)纳米颗粒。将其置于本课题组自制的VMF中,研究其介导的磁场机械力对肿瘤细胞的杀伤效果。结果单纯施加VMF对细胞活力无影响;加入Fe_(3)O_(4)纳米颗粒后,细胞活力随VMF处理时间和Fe_(3)O_(4)纳米颗粒浓度的增加而降低,受损细胞释放的乳酸脱氢酶也随磁场处理时间延长而增加。结论不规则形貌Fe_(3)O_(4)纳米颗粒在VMF下可将机械力转移到肿瘤细胞,破坏细胞结构,导致细胞死亡;所采用的VMF装置结构简单、使用安全、操作方便。所采用的磁性粒子及其杀伤肿瘤细胞的方法,有临床转化潜力。展开更多
A novel phenanthrene imine was synthesized from 9,10-phenanthrenequinone, 1,4-diazabicyclo octane （Dabco）, 2,6-dimethylaniline, 2,6-diisopropylaniline and TIC14 via standard Schlenk and vacuum-line or glovebox techn...A novel phenanthrene imine was synthesized from 9,10-phenanthrenequinone, 1,4-diazabicyclo octane （Dabco）, 2,6-dimethylaniline, 2,6-diisopropylaniline and TIC14 via standard Schlenk and vacuum-line or glovebox techniques. 3-（4,5-Dimethylthiazolyl-2）-2,5-diphenyltetrazolium bromide（MTT） assay and flow cytometry（FCM） were used to detect the activities of the compound to kill tumor cells and the mechanisms of action were investigated in the study. The results show that the compound is active in killing tumor cells. Mechanistic investigations found that the compound could induce apoptosis of tumor cells.展开更多
Objective: To study the enhancement of the immune functions and autologous tumor killing (ATK) activity by kappa selenocarrageenan (KSC) in mice bearing sarcoma 180. Methods: To measure the effects of KSC and/or Cy...Objective: To study the enhancement of the immune functions and autologous tumor killing (ATK) activity by kappa selenocarrageenan (KSC) in mice bearing sarcoma 180. Methods: To measure the effects of KSC and/or Cyclophosphamide (Cy) on natural killer (NK) activity, lymphokine activated killer (LAK) activity, the produc tion of interleukin 2 (IL 2), ATK activity and the growth of sarcoma 180 (S 180 ). Results: KSC promoted NK activity, LAK activity and ATK activity in vivo , increased IL 2 production at 40 mg/kg/d×9d. It also enhanced the antitumor action of Cy (20 mg/kg/d×9d) and offset the inhibition of Cy on immunocopetent cells. The ATK activity in splenocytes of S 180 bearing mice could be induced and increased by recombinant interleukin 2 (rIL 2) in vitro . Conclusion: KSC has an up regulating effect on the immune functions and ATK activity in tumor bearing mice. It can be used as a biological response modifier (BRM) in cancer biotherapy.展开更多
文摘From the mouse hybridoma cell line secreting an anti-CD4 monoclonal antibody (McAb), total RNA was prepared. The VH and VL genes were amplified by RT-PCR with family specific primer pairs. The PCR products were cloned into pGEM-T vectors, then tranfected into JM109. The VH and VL genes were snalyzed by automatic DNA sequencer. According to Kabat classification, the VH and VL genes belong to the mouse ig heavy subgroup Ⅱ(A) and x chain subgroupⅢ, respectively. The VH and VL genes were subcloned into pr1-Expr and Pk Expr respectively, then transfected into XL2-Blue. The VH- Pr1 and VL- pk were trans feeted by electroporation into mouse myeloma cell X63Ag8. 653. The transfectoma cells were selected by G418 screening, and then supernatant of cultured transfectoma were analyzed by ELISA and immunofluorescence techniques.We have acquired transfectoma cells secreting anti-CD4 chimeric antibodies.These chimeric antibodies are able to kill tumor cells specifically in vitro.
基金supported by the National Natural Science Foundation of China,China(82072087,31670880 and 31970893)Guangdong Natural Science Fund for Distinguished Young Scholars,China(2017A030306016 and 2016A030306004)Fundamental Research Funds for the Central Universities,China(19ykzd39)
文摘Adoptive cell therapy(ACT)is an emerging powerful cancer immunotherapy,which includes a complex process of genetic modification,stimulation and expansion.During these in vitro or ex vivo manipulation,sensitive cells are inescapability subjected to harmful external stimuli.Although a variety of cytoprotection strategies have been developed,their application on ACT remains challenging.Herein,a DNA network is constructed on cell surface by rolling circle amplification(RCA),and T cell-targeted trivalent tetrahedral DNA nanostructure is used as a rigid scaffold to achieve high-efficient and selective coating for T cells.The cytoprotective DNA network on T-cell surface makes them aggregate over time to form cell clusters,which exhibit more resistance to external stimuli and enhanced activities in human peripheral blood mononuclear cells and liver cancer organoid killing model.Overall,this work provides a novel strategy for in vitro T cell-selective protection,which has a great potential for application in ACT.
文摘目的考察Fe_(3)O_(4)纳米颗粒在低频振动磁场(low-frequency vibrating magnetic field,VMF)驱动下通过磁场机械力杀伤肿瘤细胞的效果。方法通过共沉淀法合成一种磁性强、具有不规则形貌的立方相Fe_(3)O_(4)纳米颗粒。将其置于本课题组自制的VMF中,研究其介导的磁场机械力对肿瘤细胞的杀伤效果。结果单纯施加VMF对细胞活力无影响;加入Fe_(3)O_(4)纳米颗粒后,细胞活力随VMF处理时间和Fe_(3)O_(4)纳米颗粒浓度的增加而降低,受损细胞释放的乳酸脱氢酶也随磁场处理时间延长而增加。结论不规则形貌Fe_(3)O_(4)纳米颗粒在VMF下可将机械力转移到肿瘤细胞,破坏细胞结构,导致细胞死亡;所采用的VMF装置结构简单、使用安全、操作方便。所采用的磁性粒子及其杀伤肿瘤细胞的方法,有临床转化潜力。
文摘A novel phenanthrene imine was synthesized from 9,10-phenanthrenequinone, 1,4-diazabicyclo octane （Dabco）, 2,6-dimethylaniline, 2,6-diisopropylaniline and TIC14 via standard Schlenk and vacuum-line or glovebox techniques. 3-（4,5-Dimethylthiazolyl-2）-2,5-diphenyltetrazolium bromide（MTT） assay and flow cytometry（FCM） were used to detect the activities of the compound to kill tumor cells and the mechanisms of action were investigated in the study. The results show that the compound is active in killing tumor cells. Mechanistic investigations found that the compound could induce apoptosis of tumor cells.
文摘Objective: To study the enhancement of the immune functions and autologous tumor killing (ATK) activity by kappa selenocarrageenan (KSC) in mice bearing sarcoma 180. Methods: To measure the effects of KSC and/or Cyclophosphamide (Cy) on natural killer (NK) activity, lymphokine activated killer (LAK) activity, the produc tion of interleukin 2 (IL 2), ATK activity and the growth of sarcoma 180 (S 180 ). Results: KSC promoted NK activity, LAK activity and ATK activity in vivo , increased IL 2 production at 40 mg/kg/d×9d. It also enhanced the antitumor action of Cy (20 mg/kg/d×9d) and offset the inhibition of Cy on immunocopetent cells. The ATK activity in splenocytes of S 180 bearing mice could be induced and increased by recombinant interleukin 2 (rIL 2) in vitro . Conclusion: KSC has an up regulating effect on the immune functions and ATK activity in tumor bearing mice. It can be used as a biological response modifier (BRM) in cancer biotherapy.
