| Background Th2 cell and its cytokines play important roles in the process of cancer development.IL-4 and IL-13 are important Th2 cytokines.They can promote Treg,MDSC and TAM tumor infiltration,inhibit anti-tumor immune response mediated by CTL and Thl cell,which accelerate the formation of tumor immunosuppressive microenviroment.Therefore,targeting IL-4 and IL-13 has the potentials to normalize the tumor microenvironment,stimulate the reproduction of anti-tumor effector cells,enhance host anti-tumor cellular immunity.The majority of tumors display heterogeneity.It limits the vaccine development against tumor specific antigen and achievement of remarkable clinical effect.Chemotherapy is a common method for clinical therapy.Chemotherapy drugs have the direct tumoricidal activity and release a large number of tumor antigens.In addition,Chemotherapy drugs could induce immunogenic cell death and excite host tumor immunity targeting multiple tumor-associated antigens.However,this anti-tumor immunological effect is often limited by the effect of tumor immunosuppression mechanism.Objective The purpose of this study intends to investigate the potential influence of targeting IL-4 and IL-13 through an active immunization approach on tumor growth,tumor metastasis and immune responses in mouse 4T1 Breast tumor model.Meanwhile,explore the potential cooperative action of the combination application of chemotherapy drugs and active intervention by targeting IL-4 and IL-13.This study can help us understand and disclose the mechanism of the tumorigenesis of IL-4 and IL-13 and provide innovative tactics for cancer immunotherapy.Methods The recombinant plasmid encoding for a chimeric protein with IL-4 or IL-13 antigen epitope inserted into the immunodominant epitope of HBcAg,was transformed into E.coli DH5 a cells,and the expression of the recombinant proteins were induced by IPTG.The proteins were purified and the presence of VLPs was detected with electron microscopy.Mice were immunized with the VLPs through preventive strategies in the breast cancer metastatic mouse models and in-situ model,in order to study the influence on tumor growth and metastatic and to explore immune response characteristics.Moreover,intratumor injection with chemotherapy drugs mitoxantrone in the in-situ transplanted tumor model were carried out when the tumor reached 5-6mm.Tumor size was measured every three or four days and mice were sacrificed at the end of the experiment.The tumor was weighed and the number of lung nodes were recorded.Flow cytometry was used to analyze the response level and function of immune effector cells and immunosuppressive cells.Results The recombinant protein was expressed efficiently in E.coli,and the purified protein presents mainly in the form of VLPs.Immunization with VLPs induced high titers of specific IgG.The growth and metastasis of tumor were suppressed in the breast cancer metastatic mouse models and in-situ model,an IL-4/IL-13 dual targeting had more remarkable limiting effect as well.Analyzed by flow cytometry,the level of CTL cell and Thl cell in splenocytes were increased,while the number of Th2 cell,MDSC and Treg in splenocytes were decreased.While targeting IL-4 or IL-13 immunization was combined with the chemotherapy drugs,significant inhibitive effects were found,indicating synergistic reaction occurred.Conclusion Targeting IL-4 and IL-13 through an active immunization approach has the potentials to normalize the tumor microenvironment and inhibit tumor growth and metastasis.Meanwhile,when it is combined with the chemotherapy drugs,significant inhibitive effects were found,indicating it has promising potential as an attractive target for immune intervention in cancer,especially combined with other tumor therapy methods. |
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