Study On Immunological Effect Of Dendritic Cell Transfected By Recombinant Plasmid Of ΔNp73α On Breast Cancer

ObjectiveTo research the immunological effect of dendritic cell transfected by recombinant plasmid of ΔNp73α on breast cancer.MethodsCultivate dendritic cells by cytokines(rh GM-CSF、rh IL-4、rh TNF-α) from healthy human umbilical cord blood in vitro, CD1 a,CD83 were detected by flow cytometry. Recombinant plasmid ΔNp73α was used for transfecting DCs, the expression of protein was detected by Western blot. Transfected DCs cocultivate with T cell induced specific cytotoxic T lymphocytes(CTL). Changes of lymphocyte subsets before and after cocultivated were detected by FCM. The secretion of IFN-γ level in the culture supematant were detected by ELISA. The killing effect on MDA-MB-231 cell was detected by LDH method. Using SPSS17.0 software for statistical analysis of experimental data.Result1.MNC derived from umbilical cord blood could be induced to mature in the medium containing rh GM-CSF、rh IL-4、rh TNF-α.2.The expression of CD1a(58.78%),CD83(76.61%)in mature DC was higher than CD1a(20.34%) and CD83(15.13 %) in immature DC((P<0.01).3.Western blot was used to detect the expression of ΔNp73α protein in the transfected DC and it had shown a special band.4.CD4+T lymphocyte in T-DC group and T-DC-ΔNp73α group were(29.12±0.61)%、(31.52±0.97)%,CD8+T were(18.82±1.29)%、(45.71±0.74)%, which were higher than T lymphocyte group(18.31±1.81)%、(18.59±2.18)%, and CD4+/CD3+ in T-DC group and T-DC-ΔNp73α had no statistical significant(P>0.05), but CD8+/CD3+ in T-DC group and T-DC-ΔNp73α were significant difference(P<0.01).5.The killing effect of specific CTL induced by ΔNp73α-DC and T cells cocultivation on breast cancer cell MDA-MB-231 was statistic stronger than DC group(P<0.05), the secretion level of IFN-γ compared with DC and pc DNA group were statistically significant(P<0.01).Conclusion1.Cultivated successfully mature dendritic cells by cytokines from MNC. And ΔNp73α obtained expression in DC cell. They both illustrate the feasibility on preparation of ΔNp73α-DC vaccine.2.The CTL induced by ΔNp73α transfected DC vaccine has significant effect on killing breast cancer. Further illustrate the effective on preparation of ΔNp73α-DC vaccine.