The Investigation Of Liposomal Medicine And Biological Adjuvants In Cancer Therapy

Objective To explore anti-tumor effects and toxicity responses of pegylated liposomal daunorubicin(PL-DNR) for curing leukemia in mouse model.Methods PL-DNR was prepared using methods of dry lipid hydration and remote loading,then physiochemical index was analysed.The in vitro cytotoxicity experiment and the in vivo therapeutic trial using animal model were performed.The apoptotic cardiomyocytes were detected using the terminal deoxynucleotidyl transferase mediated dUTP nick end labeling method(TUNEL).Results Average diameter of PL-DNR was(110±10)nm.The encapsulation efficiency was 94.21%.The growth of tumors in PL-DNR and DNR therapy group was obviously suppressed,which reached 77.59%and 57.56%respectively with a significant difference between them (P＜0.05).Compared with PL-DNR group,DNR group had significantly higher index of apoptotic cardiomyocytes(P＜0.05).The same results can be obtained in the therapeutic models of P388 and L615.Conclusion In treatment of the leukemia three models in mice,pegylated liposomal daunorubicin can improve therapeutic effects and reduce cardiomyocyte apoptosis in daunorubicin-induced cardiomyopathy. Objective poly(I:C) and CpG ODN can activate immunological reactions respectively.In the experiment,we combinate CpG ODN and poly(I:C) as the adjuvant of tumor associated antigen MUC-1 to oberserve whether enhanced stimulation of innate immunological response and adaptive immunological response can be induced.Methods In vitro,murine splenocytes were incubated with poly(I:C) alone,CpG ODN alone or the combination of the two agents.Then the levels of cytokine production of splenocytes were examined including Interleukin(IL)-12p40, tumor necrosis factor-α(TNF-α),IFN-γand IL-4 by the method of ELISA.In vivo,to detect if there is synergetic effect of the combination of two immunological stimulator as adjuvant of MUC-1 peptide,C57BL/6 were immunized with MUC1 plus poly(I:C) and/or CpG ODN.Then the serum levels of cytokines were detected including.We use the CTL test to evaluate the cellular immune response,and use the serum level of MUC-1 antibody to evaluate the humoral immune response.At last we ultilize several mice to produce the monoclonal antibody of MUC-1.Results In vitro,the combination of CpG ODN and poly(I:C) demonstrated synergetic effect for the increased levels of IL-12p40,TNF-αproduction.The same result can be observed in vivo experiment,that is,immunization of mice with the combination of poly(I:C) and CpG ODN demonstrated synergy for the increased serum levels of IL-12p40,TNF-α. Then we obtained a strain of monoclonal anti-MUC-1 antibody with high titer. Conclusions The combination of poly(I:C) and CpG ODN as the adjuvant of peptide MUC1,resulted in an enhanced innate and adaptive immune response that could be used for tumor vaccination or immunotherapy.