The Antitumor Effect And Mechanism Research Of Fusion Hybrid Of Dendritic Cells And Myeloma Cell SP₂/0 Expressing CD44v6

Objective:To study the antitumor effect and mechanism of fusion hybrid of dendritic cells and myeloma cell SP2/0 expressing CD44v6, the dendritic cells(DCs) and Myeloma Cell SP2/0 derived from mice were fused, and modified with CD44v6, it will provide the experimental and theoretical foundation on the immunotherapy for tumor patients.Methods : (1)To constructed for recombinant expression vector expressing CD44v6, at first the gene encoding CD44v6 was amplified from human cancer cells by RT-PCR, and inverted in to the eukaryon expression vector–pBud, the recombinant vector pBud-CD44v6 was transformed into E.coli DH5αand identified by Enzyme digestion and DNA sequencing analysis; (2)Bone marrow-derived mice DCs were largely expanded with cytokine of GM-CSF, IL-4 and TNF-2; (3)DCs and SP2/0 cells were fused by polyethylene glycal(PEG) and the completely fused ones were screened by HAT/HT system; (4)The recombinant pBud-CD44v6 vector was transfected into the fused cells (hybridoma cell) by Lipofectamine 2000, and identified by RT-PCR (5)The anticancer effect and cytotoxicty of fused–cell vaccine were studied in vitro; (6)Fused cells were implaned to bearing-cancer mice, and the volume of tumors and life span of mice were studied.Results:DC/SP2/0 fused cell vaccine modified with CD44v6 was successfully constructed; The cytotoxicity T lymphocyte(CTL) of immune group was larger than that of the control group, the volume of tumors were smaller than that of control ones, and the life span of immune group was extended.Conclusion:Mice Bone-marrow derived DC cells can be fused with SP2/0 cells by PEG, and the fused cells can induce strong antitumor immune effect of T lymphocyte.