CAJ | 학술논문

目的：优化大肠杆菌菌蜕装载质粒的效率，并将装载质粒的菌蜕转染抗原提呈细胞，以提高核酸疫苗的递送水平。方法：将质粒pHH43转化大肠杆菌DH5α，制备大肠杆菌菌蜕；优化菌蜕装载质粒时菌蜕、质粒和膜囊的比例，获得更高的装载效率，通过扫描及透射电镜、流式细胞术观察其形态变化及装载效率；将装载质粒的菌蜕与抗原提呈细胞--巨噬细胞RAW264.7和树突状细胞DC2.4共孵育，观察吞噬效果。结果：优化了大肠杆菌菌蜕装载质粒的效率，当菌蜕、质粒、膜囊的比例为7∶10∶4时效率达到最佳，装载DNA效率达98%以上；抗原提呈细胞吞噬装载了质粒的菌蜕，效率达100%。结论：大肠杆菌菌蜕可高效装载核酸疫苗，且高效被抗原提呈细胞捕获，有助于提高核酸疫苗的递送和免疫效果的提高。
목적：우화대장간균균세장재질립적효솔，병장장재질립적균세전염항원제정세포，이제고핵산역묘적체송수평。방법：장질립pHH43전화대장간균DH5α，제비대장간균균세；우화균세장재질립시균세、질립화막낭적비례，획득경고적장재효솔，통과소묘급투사전경、류식세포술관찰기형태변화급장재효솔；장장재질립적균세여항원제정세포--거서세포RAW264.7화수돌상세포DC2.4공부육，관찰탄서효과。결과：우화료대장간균균세장재질립적효솔，당균세、질립、막낭적비례위7∶10∶4시효솔체도최가，장재DNA효솔체98%이상；항원제정세포탄서장재료질립적균세，효솔체100%。결론：대장간균균세가고효장재핵산역묘，차고효피항원제정세포포획，유조우제고핵산역묘적체송화면역효과적제고。
Objective: To optimize plasmid DNA loading procedure of E.coli bacterial ghosts(BG), and to trans-fect the resulting BG into antigen presenting cells(APC) to improve the delivery efficiency of DNA vaccine. Methods: Plasmid pHH43 was transformed into E.coli and the E.coli BG was prepared by lysis E protein in a temperature controlled manner. The plasmid DNA were loaded into E.coli BG and for higher efficiency, the ratios of BG, plasmids,and membrane envelope were optimized during the DNA loading. Then BG with plasmid were ob-served and detected by transmission electron microscope, scanning electron microscope and FACS. BG loaded with plasmids were incubated with APC human macrophage cells RAW264.7 and human dendritic cells DC4.2. Re-sults: The efficiency of E.coli loading plasmid was improved to more than 98% when the ratio of BG, plasmids, and membrane envelope was 7∶10∶4. The BG loaded with plasmid DNA could be observed in almost all APC. Conclusion: The efficiencies of E.coli BG loading DNA plasmid and subsequent transfecting APC were high so that DNA vaccine delivery can be accomplished.