Abstract
Objective To observe the immune effect of dendric cell transduced with breast cancer specific gene 1 (Bcsg1) and to conduct safety evaluation. Methods Dendritic cells originated from the peripheral blood mononuclear cells were transfected with recombinant adeno-associated virus (rAAV) containing Bcsg1 full length cDNA to generate Bcsg1 gene modified DC vaccine T lymphocytes. Those genetically modified DC vaccine repeatedly generate Bcsg1 specific cytotoxicity T lymphocytes (CTL). CTL-mediated cell lysis to mammary cancer cells line Bcsg1 (+ )-SKBR-3. The surface molecular of mature DC cells and CTL activation markers were detected by Flow cytometry. The secretion levels of interferon (IFN)-γ and interleukin (IL)-4 in CTL activated by rAAV/Bcsg1-DC, Lysate-DC and n-rAAV/Bcsg1-DC were detected by flow fluorescence. rAAV gene residues were detected in mature CTL cells and culture fluid of those cells. Results After transfection, the expression of CD80 (90.34±1.57), CD83 (85.14±1.54), CD86 (82.36±1.38), human leukocyte antigen-DR (HLA-DR) (78.16±1.13) molecular markers of DC were significantly different from the control group (P=0.000) and induced CTL activation was significantly increased. In the course of CTL induction, IFN-γ [(2 616.53±163.81) pg/ml] and IL-4 [(437.36±46.33) pg/ml] levels were significantly different from the control group (P=0.000). And rAAV/Bcsg1-DC-CTL has a stronger cytotoxic activity and higher targeting property. When the target ratio was 40∶1, the killing rate of target cells was (85.40±8.48)% and on non-target cell killing rate is only (24.50±4.15)%. rAAV gene residues were not detected in mature CTL cells and culture fluid. Conclusion rAAV/Bcsg1 transfected DC activate mammary cancer specific CTL. It is a safe and effective method for the treatment of tumor immune gene therapy. Key words: Adenoviridae infections; T-lymphocytes; Cytotoxic; Cancer vaccines; Breast neoplasms; Immunotherapy
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