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Objective:To investigate effects of tumor specific TCR gene Vα12.2-Vβ7.1 modification on recognition of tumor antigen and activation of anti-tumor reactivity of T cells.Methods: T cells were transduced using recombinant Ad 5F35-TRAV-TRBV adenovirus ,and multiplicity of infection was optimized.Specific lysis of T cells was evaluated by calcein release assay.The frequency of apoptotic cells in target cells was detected by Annexin V /PI double-labeled FACS.The expression of FasL on T cells was analyzed by FACS.The secretion of cytokine IFN-γand IL-2 of T cells was determined by ELISA assays.Results: The highest tranduce efficiency was obtained at MOI 100 by recombinant Ad5F35-TRAV-TRBV adenovirus.The frequency of TCRVα12+Vβ7+cells reached above 25%3 days after transduction.TCR gene modification enhanced the ability of T cells to lyse HLA-A2+AFP+target cells(P<0.001), the ability of T cells to induce HepG-2 apoptosis(P<0.001),and expression of FasL on T cells(P<0.001).TCR gene modification also enhanced T cells to secret IFN-γafter coculture with antigen positive tumor cells ( P<0.001 ).Conclusion: Specific TCR gene modification by recombinant adeno virus effectively promotes T cells to recognize antigen positive tumor cell and exert anti -tumor reac-tivity.
RESUMO
Aim By establishing mouse acute myocardial infarction model,to observe the protection of isoflavones on ischemic myocardium and research the mechanism.Methods Mouse acute myocardial infarction model was established by ligating the left anterior descending(LAD)coronary artery.Danshen was used as the positive control.The effect of isoflavones on myocardial infarct area,serum myocardium creatase and serum levels of SOD and MDA was observed.By Real Time PCR,it was found that isoflavones could affect the expression of β-adrenergic receptor kinase(β-ARK_1).Results Isflovones could obviously reduce the myocardial infarct area and lower the levels of serum myocardial creatase and MDA.It could downregulate the expression of β-ARK_1 as the doses are increased.Conclusions Isoflavones can protect the myocardium of acute myocardial infarction mouse.The mechanism is related to the reduction of the oxidative damage,and the downregulation of the expression of β-ARK_1.
