Research On Immunologic Adjuvant Effects Of Cinobufagin And Its Enhancement Function Of Tumor-specific Immune Response

Cinobufagin belongs to the bufadienolide components, is one of the effective activeingredients in Venenum Bufonis. As a single component, cinobufagin has a clear molecularstructure and become a hotspot in the field of pharmaceutical research at home and abroad.Extensive literature has confirmed cinobufagin to have anti-tumor, immunological enhancement,regulating the cardiovascular system, analgesics, anti-virus and other effects. But the effects ofcinobufagin in tumor immunotherapy have not been reported. This research aimed to explore theimmunologic adjuvant effects of Cinobufagin and its enhancement function of tumor-specificimmune response.In this study, we first immunized DBA/2J mice with P815AB peptide or Cinobufagin. Toobserve the splenic index and thymus index, MTT assay was used to detect the proliferation of thesplenic lymphocytes. Flow cytometry (FCM) was used to observe T lymphocytes subsets inperipheral blood. ELISA was used to detect the secretion of IFN-γ in peripheral blood. At the sametime, the activity of CTL and the phagotrophic activity of mice macrophage were alsoobserved.On the other side, we also detected the expression of MHC-Ι and genes LMP2, LMP7,TAP1and TAP2in B16cells and RMA-S cells by FCM and real-time fluorescent quantitativePCR.The results showed that the proliferation of the splenic lymphocytes. The activity of CTL, thephagotrophic activity of mice macrophage, the percent of CD3+CD4+CD8+T lymphocytes andCD3+CD69+T lymphocytes in peripheral blood were increased remarkably in the DBA/2J miceimmunized by P815AB peptide and Cinobufagin together, yet the viscera index and the secretionof IFN-γ in peripheral blood were increased slightly. Additioally, the mRNA expression of genesLMP2, LMP7, TAP1, TAP2were increased remarkably by60μg/mL Cinobufagin while theexpression of MHC-Ⅰ was not affected significantly by Cinobufagin neither in B16cells norRMA-S cells,. The results suggest that Cinobufagin has immunologic adjuvant effects and the potential to enhance the immunogenicity of tumor by means of increasing the expression of LMPand TAP.