摘要
Background/Objective: Recently, cellular immunotherapy research of leukemia is being widely explored. However, how to develop a simple, safe and effective way to stimulate immune cells and activate anti-tumor immunity still need further investigating. DC driven cold thermal plasma, which creates a variety of active gas components and almost no injury to tissue cells, has emerged in the biomedical field including food disinfection, wound healing, surgical procedures and cancer treatment. The plasma treatment may become a new cellular immunotherapy treatment other than conventional chemotherapy. Nevertheless, the effect of cold thermal plasma in leukemia immunotherapy and the possible mecha-nism remains unclear. Here, we investigated the mechanism of cold thermal plasma enhances anti-leukemia effects which may improve the research basis for immunotherapy of leukemia. Methods: The co-culture model of macrophage cell line RAW264.7 and leukemia cell lines L1210 was established, cold thermal plasma stimulation time gradient was set by 0s,20 s,40s,60 s,80s and 100 s. Macrophage phagocytosis ability was analyzed by neutral red staining, and enhanced phagocytosis cells were counted by flow cytometry. Cytokines of TNF-α,TGF-β, IL-1β, IL-6 and IL-10 secreted in culture supernatant were measured by ELISA, and real-time PCR and Western-Blot were used for detecting the mRNA and protein level of these cytokines in macrophage. Result: Neutral red staining and flow cytometry showed that phagocytosis of macrophages stimulated by cold thermal plasma were enhanced, compared with the control group(P<0.05), and saturated by 98.6% at 80 s stimulation. Cell coculture experiments showed that the survival rate of leukemia cells was decreased after stimulation of cold thermal plasma. Furthermore, the expression of TGF-β, IL-10 was significantly increased, consistent with the results of real-time quantitative PCR and Western-Blot. Conclusion: Our finding reveals that cold thermal plasma can enhance phagocytosis of macrophage and anti-leukemia effort.
Background/Objective: Recently, cellular immunotherapy research of leukemia is being widely explored. However, how to develop a simple, safe and effective way to stimulate immune cells and activate anti-tumor immunity still need further investigating. DC driven cold thermal plasma, which creates a variety of active gas components and almost no injury to tissue cells, has emerged in the biomedical field including food disinfection, wound healing, surgical procedures and cancer treatment. The plasma treatment may become a new cellular immunotherapy treatment other than conventional chemotherapy. Nevertheless, the effect of cold thermal plasma in leukemia immunotherapy and the possible mecha-nism remains unclear. Here, we investigated the mechanism of cold thermal plasma enhances anti-leukemia effects which may improve the research basis for immunotherapy of leukemia. Methods: The co-culture model of macrophage cell line RAW264.7 and leukemia cell lines L1210 was established, cold thermal plasma stimulation time gradient was set by 0s,20 s,40s,60 s,80s and 100 s. Macrophage phagocytosis ability was analyzed by neutral red staining, and enhanced phagocytosis cells were counted by flow cytometry. Cytokines of TNF-α,TGF-β, IL-1β, IL-6 and IL-10 secreted in culture supernatant were measured by ELISA, and real-time PCR and Western-Blot were used for detecting the mRNA and protein level of these cytokines in macrophage. Result: Neutral red staining and flow cytometry showed that phagocytosis of macrophages stimulated by cold thermal plasma were enhanced, compared with the control group(P<0.05), and saturated by 98.6% at 80 s stimulation. Cell coculture experiments showed that the survival rate of leukemia cells was decreased after stimulation of cold thermal plasma. Furthermore, the expression of TGF-β, IL-10 was significantly increased, consistent with the results of real-time quantitative PCR and Western-Blot. Conclusion: Our finding reveals that cold thermal plasma can enhance phagocytosis of macrophage and anti-leukemia effort.
引文