Silenced SOCS1in DC Loaded With Specific Antigen Enhances The Effect Of Anti-multiple Myeloma By T Cell Immunity

Objective:To improve the dendritic cell abilities of antigen presentation and maturation and to enhance anti-tumor immunity in vivo by Silencing of SOCS1with miRNA.Methods:1MiRNA Oligo was pulged in eukaryotic expression vector pcDNA6.2-GW/EmGFP-miR by BLOCK-iTTM Pol miR RNAi Expression Vector Kit with EmGFP2Plasmid DNA was amplified and extracted by Fastfilter endofree Plasmid Maxi Kit.3Bone marrow progenitor cells were prepared from BALB/c mice and cultured with GM-CSF and IL-4to generate DC. DC was detected by Cytomorphology, MLR and Flow Cytometry.4Vetors were transfected into DC with LipofectamineTM LTX Transfection Reagent. Expression level of SOCS1in DC was detected by western blot.5Pyrolysis product was prepared from the cultured P3X63Ag8.653multiple myeloma cellls6Animal models of tumor had been established by injection.7The animals in the models of tumor were immunized with multiple myeloma cells which were the pulsed, socsl-silenced mature DCs; Levels of IFN-r of Spleen lymphocytes were deteted by ELIS A after immunization; The cytotoxic T lymphocyte activity was detected by the method of LDH; Sizes of tumors were measured and the inhibitition of tumor groth was observed.Results:1The recombinant plamid vctors pcDNA6.2-GW/EmGFP-miR were constructed successfully.2DC can be measured by Cytomorphology, MLR and Flow Cytometry.3The downregulated level of socsl detected in DC by Western blot4Antigen-pulsed, socsl silenced mature DC Cells can induce more strongy response of CTL killing of multiple myeloma cellls and inhibit groth of animal tumor.Conclusions:Antigen-pulsed, socsl silenced mature DC Cells may enhance anti-tumor immunity and inhibit groth of animal tumor...