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【目的】探索基于pH值敏感的荧光染料分析腺病毒裂解T淋巴细胞胞内体膜的实验方法。【方法】本文以Jurkat细胞(T淋巴瘤细胞)为靶细胞,将pH值敏感的荧光染料pHrodo dextran与5型腺病毒(Ad5)共同孵育Jurkat细胞,对pHrodo dextran孵育的浓度与时间进行了优化,利用激光共聚焦显微镜分析胞内相对平均荧光强度百分比随时间的变化情况,反映Ad5诱导胞内体膜裂解情况。【结果】研究结果表明,在pHrodo dextran终浓度为80μg/m L,孵育时间为10 min条件下,在病毒感染后的30 min,相对平均荧光强度百分比出现显著下降;利用巴佛洛酶素A1抑制胞内体膜质子泵活性后,相对平均荧光强度百分比出现轻微下降。【结论】建立了基于pHrodo dextran分析腺病毒诱导T细胞胞内体膜裂解的新方法。
【Objective】 The objective of this study was to explore an experimental method for the analysis of adenovirus lysed T lymphocytes intracellular membranes based on pH-sensitive fluorochromes. 【Method】 Jurkat cells (T lymphoma cells) were used as target cells to incubate Jurkat cells with pHrodo dextran and adenovirus type 5 (Ad5). The concentration and time of pHrodo dextran incubation were optimized , Using confocal laser scanning microscope intracellular relative fluorescence intensity percentage changes over time, reflecting the endometrial membrane Ad5-induced cleavage. 【Result】 The results showed that the relative average fluorescence intensity decreased significantly after 30 min of the infection with pHrodo dextran at a final concentration of 80 μg / mL and incubation time of 10 min. The inhibition of endosomal membrane proton pump activity, the relative average fluorescence intensity percentage decreased slightly. 【Conclusion】 A new method of adenovirus-induced endosomal membrane lysis by T cells was established based on pHrodo dextran assay.