人树突状细胞与肺癌细胞A-549融合疫苗生物学特性研究

Vaccine research on biological characteristics of human dendritic cells and A-549 lung cancer cell fusion

目的探讨人树突状细胞(DC)与人肺癌细胞A-549融合所得疫苗在制备过程中的生物学特性,总结高效制备融合疫苗的方法。方法应用GM-CSF和IL-4优化的方法制备肺癌患者人外周血单核细胞以获得DC,寻找DC制备率最高时间段;同时应用PKH672GL(绿色荧光)和PKH262GL(红荧光)分别标记DC和肺癌细胞A-549细胞,筛查最佳的融合比例。结果应用GM-CSF和IL4优化法进行DC制备第7天所得百分率为(66.26±5.13)%,高于其他时间(P<0.05);通过对比不同融合比例DC与人肺癌细胞A-549,显示1:1时所取得的融合百分率为(35.15±2.16)%,高于其他比例(P<0.05)。结论在DC制备过程中制备第7天所得DC百分率最高,应选取此时作为提取DC的最佳时间;同时DC与人肺癌细胞A-549以1:1比例相融合所得疫苗百分率最高。

Objective To explore the human dendritic cells （DC） and A-549 in human lung cancer cell fusion vac -cine in the biological characteristics of the process of preparation , summarize the methods of efficient preparation of fusion vac -cine.Methods By using of GM-CSF and IL-4 optimization method for preparing patients with lung cancer in human peripher -al blood mononuclear cells for DC, DC looking for the highest rate of preparation time ; while applying PKH672GL （green flu-orescence） and PKH262GL （red fluorescence） and lung cancer cells were labeled DC and A -549 cells, screening the best blend ratio.Results Application of GM-CSF and IL-4 optimization method for the first 7 days resulting percentage was （66.26 ＆#177;5.13）%, higher than at other times （ P ＆lt;0.05） DC preparation; By comparing different fusion the proportion of DC with human lung cancer cell A -549, showing the percentage of fusion was obtained （35.15 ＆#177;2.16）%, higher than the other ratios （ P ＆lt;0.05）.Conclusion The seventh days ＆#39;percentage of DC is the best in the preparation process of DC , should be selected as optimum extraction time DC ; while DC and human lung cancer cell line A -549 with a ratio of 1∶1 shows the highest percentage combined vaccine .