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Study On Therapeutic Effect Of Whole Cell Vaccine On Melanoma

Posted on:2023-09-27Degree:MasterType:Thesis
Country:ChinaCandidate:Z ZengFull Text:PDF
GTID:2544306833953199Subject:Special medicine
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Objective:Melanoma is a fatal skin malignancy,and its incidence rate continues to rise worldwide.The traditional methods for the treatment of malignant melanoma,such as surgery,radiotherapy and chemotherapy,have the problems of great trauma and obvious adverse reactions,and can not significantly improve the survival rate of patients with advanced malignant melanoma.In recent years,biological immunotherapy has made a breakthrough in the treatment of malignant melanoma.Among them,tumor cell vaccine has attracted much attention as a new way of tumor treatment.As a new way of tumor treatment,tumor cell vaccine has attracted much attention.However,the immunogenicity of tumor cell vaccine alone is poor,so we use laminarin sulfate and UV inactivated tumor cell vaccine to treat mouse melanoma,and evaluate its therapeutic effect through in vivo and in vitro experiments,so as to provide a possible choice and idea for clinical tumor immunotherapy from the perspective of basic experiments.Methods:The cell vaccine was prepared by inactivating mouse melanoma cells by UV irradiation.The death of melanoma cells was detected by trypan blue staining,and the proliferation of melanoma cells was detected by CCK8 method.In order to verify the therapeutic effect of cell vaccine combined with laminarin sulfate on melanoma,42C57BL/6 mice were randomly divided into 7 groups with 6 mice in each group,and1*10~5melanoma cells were injected subcutaneously into the back.PBS,whole cell vaccine,aluminum adjuvant,laminarin sulfate,whole cell vaccine combined with aluminum adjuvant and whole cell vaccine combined with laminarin sulfate were injected into the thigh muscle of tumor bearing mice on day 3 and day 10 respectively.Another group of mice injected whole cell vaccine combined with laminarin sulfate into the thigh muscle of tumor bearing mice on day-7 and day 10 respectively.The tumor length and width of tumor bearing mice was measured and recorded every other day from the 9th day.The survival status of mice in each group,such as hair color change,weight,diet,water intake and activity,were observed.The changes of T cells and cytokines in each group were detected by flow cytometry.Results:1.Trypan blue staining and CCK8 results showed that UV irradiation for 10 min caused melanoma cells to die and maintain intact morphology.2.The whole cell vaccine combined with Laminaria polysaccharide sulfate group (prevention+treatment)significantly delayed the growth rate of melanoma,and the tumor volume of this group was significantly smaller than that of the control group (P<0.05).3.Compared with the control group,the whole cell vaccine combined with Laminaria polysaccharide sulfate group(prevention+treatment)significantly increased the TNF-αand IFN-γsecreted by CD4~+T and CD8~+T cells in the spleen of mice(P<0.05).4.Compared with the control group,the whole cell vaccine combined with Laminaria polysaccharide sulfate group(prevention+treatment)significantly increased the proportion of CD8~+T cells in tumor tissue and peripheral blood of mice(P<0.05).Conclusions:1.UV inactivated melanoma cell vaccine has good safety and immunogenicity.2.The combination of UV inactivated melanoma cell vaccine and Laminaria polysaccharide sulfate can inhibit the growth rate of melanoma.3.UV inactivated melanoma cell vaccine combined with Laminaria polysaccharide sulfate can activate the activation of T cells and cytokine secretion in mouse spleen,and up regulate its anti-tumor immune function.
Keywords/Search Tags:whole cell vaccine, adjuvant, melanoma, Immune cells, cytokine
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