Eliciting Regression By A Combination Vaccine In A Glioma Mouse Model

Glioma is the most common primary malignant brain tumor.Although advances in radiotherapy and chemotherapy have brought modest improvements over the past century,the invasive nature of the disease continue to limit the 5-year survival of glioblastoma and its variants to only 4.7%.Glioma is still an ideal target for immune-based therapy.In order to develop an efficient immunotherapy for glioma,we explored a vaccine,which combined with dendritic cells,lysate of GSC,and immunologic adjuvant CpG-ODN.An intracranial glioma mouse model was established.Afterwards,the anti-tumor effect was evaluated in glioma mouse models.Collectively,our study provided a potential GSCs therapy,and devoted to glioma immunotherapy developments.Such efforts will provide the therapeutic scheme for overcoming resistance to current treatments and preventing recurrence.Methods:1.To establish the glioma mouse model,GL261-GFP-Luc cells were implanted stereotactically into the brain using stereotaxic apparatus;2.The preparation of novel vaccineTo obtain GSCs,GL261 cells were resuspended in serum-free medium for 20 days.Then these GSCs were prepared respectively by three freeze-thaw cycles.DCs were obtained from bone marrow of C57BL/6 mice,and then cultured and extracted.CpG-ODN was dissolved in water.At last,the lysate was mixed with DCs and CpG-ODN.3.The glioma mouse model was subcutaneously vaccinated with the novel vaccine.Tumor growth and survival time were recorded.Lymphocytes were detected by flow cytometry.Conclusion1.The glioma mouse model was established successfully after GL261-GFP-Luc cells implanted into the brain.The tumor could be observed by fluorescence imaging technique;2.The vaccine treated group showed remarkable tumor growth inhibition,prolonged survival and elevated lymphocyte infiltration in glioma microenvironment.3.The vaccine including GSC lysate was more efficient than GL261 lysate in tumor eradication;4.Vaccination reduced the ratio of Treg and MDSC in tumor microenvironment,and improved the ratio of CD8+T cell.5.The results of experiments in vitro showed that the anti-tumor specific T cells were induced and cytotoxic to glioma cells.