Preparation Method Of Polyphenol-aluminum Coordination Microparticle Vaccine Using Tumor Cells As Template And Tumor Immunotherapy Effect

Aims:To develop a potent cancer vaccine,it is important to study how to prepare highly immunogenic antigens and enhance the effective uptake of tumor antigen by antigen presenting cells.But pure tumor antigen is poorly immunogenic.In order to improve the therapeutic effect of vaccines,the use of a suitable loading system is very important for enhancing antigen uptake and presentation,especially indution of cellular immune response.In this paper,a microparticles system containing tumor cell lysates was constructed by complexation of polyphenol-metal with a single cell as a template,which can effectively stimulate the presentation of tumor antigens by antigen-presenting cells and activate immune response.It provides a universal,simple and effective tumor cell encapsulation strategy that can be used for individualized immunotherapy.Method:The structure of TCL@EGCG/AIn was observed by transmission electron microscopy,scanning electron microscopy and confocal microscopy.The components of the microparticles were detected by X-ray photoelectron spectroscopy(XPS)and energy dispersive spectrometer(EDS).The protein loading capacity and pH-responsive were determined by BCA.The Confocal microscopy and flow cytometry were used to qualitatively and quantitatively analyze the microparticle uptake by DC cells;and migration of TCL@EGCG/AIn in vivo was observed by small animal imaging technology;the effect of microparticles on promoting BMDCs maturation and antigen presentation was tested by FACS for expression of surface marker.Four different animal models were used for testing the anti-tumor effect of this microparticles.Result:The microparticles of TCL@EGCG/AIn were successfully prepared and its morphological structure was determined to be spherical.It has high protein loading capacity and pH responsive and is released faster at pH 5.5.The results of cellular toxicity test show this microparticles has good biocompatibility.And the microparticles can be taken up by BMDCs cells and stimulate DC cell maturation.6 h after subcutaneous injection,the antigen was taken up by DC cell and migrated to the lymph nodes.Animal experiments have shown that it can prevent and inhibit tumor growth and metastasis to some extent.