论文部分内容阅读
目的:比较验证重组腺相关病毒负载PSA基因转染方式与LNCaP细胞裂解物诱导方式对产生树突状细胞(DCs)疫苗的的影响.方法:抽取HLA表型为A2的健康志愿者外周血,分离单核细胞,体外培养.通过反复冻融LNCaP细胞、提取肿瘤细胞裂解物以及使用含PSA片段的重组腺相关病毒转染未成熟DCs,诱导特异性细胞毒性T细胞(CTL).检测体外培养的DCs和CTL活性,并使用MTT法检测两组CTL对LNCaP细胞的杀伤作用,同时使用腺相关病毒转染组诱导的CTL对LNCaP和DU145两种前列腺癌细胞进行杀伤实验检测.结果:两种方式均可培养成熟的DCs,诱导激活CTL并分泌IFN-γ,而腺相关病毒转染的DCs成熟度和CTL诱导率均高于细胞裂解物组;腺相关病毒转染组诱导的CTL对LNCaP细胞的杀伤率显著高于细胞裂解物组诱导的CTL;腺相关病毒转染组诱导的CTL对LNCaP细胞有特异性杀伤作用.结论:两种抗原递呈方式均可培养出成熟的DCs,并可诱导自体CTL增殖;而使用重组腺相关病毒转染PSA至DCs的方式明显优于肿瘤细胞裂解物的抗原负载方式,具有高效特异性.
Objective: To compare the effect of recombinant adeno-associated virus (Adenovirus) loaded PSA gene transfection with LNCaP cell lysate on dendritic cell (DCs) vaccine.Methods: The peripheral blood of healthy volunteers with HLA phenotype A2, Monocytes were isolated and cultured in vitro, and specific cytotoxic T cells (CTLs) were induced by repeated freezing and thawing of LNCaP cells, extraction of tumor cell lysates, and transfection of immature DCs with recombinant adeno-associated virus containing PSA fragments. DCs and CTL activities were detected by MTT method and cytotoxicity of CTLs on LNCaP cells were detected by MTT assay and CTLs induced by adeno-associated virus were used to test the cytotoxicity of LNCaP and DU145 on two kinds of prostate cancer cells.Results: Mature DCs could be induced to induce CTL and IFN-γ secretion, while the maturation and CTL induction rate of DCs transfected with adeno-associated virus were higher than that of cell lysate group. CTLs induced by adeno-associated virus transfection group had no effect on LNCaP Cell killing rate was significantly higher than the cell lysate group induced CTL; adeno-associated virus-transfected group CTL induced specific killing of LNCaP cells.Conclusion: Both antigen presentation methods can be cultured into The DCs, can induce proliferation of autologous CTL; the use of a recombinant adeno-associated virus transfection embodiment PSA antigen to DCs was superior to the manner of loading of tumor cell lysate with high specificity.