微颗粒介导人乳头瘤病毒感染巨噬细胞的实验研究

发布时间：2018-08-24 14:33

【摘要】：目的：探讨人乳头瘤病毒感染巨噬细胞的途径。方法：1.从尖锐湿疣组织（CA）中提取的微颗粒和从健康人外周血中分离的巨噬细胞共培养，收集培养后的巨噬细胞并提取上清液中的微颗粒，上清液中提取的微颗粒再与新分离的巨噬细胞共培养，如此进行四次共培养，通过RT-PCR的方法检测每次共培养的巨噬细胞基因组中是否存在HPV-DNA；2.免疫细胞化学方法观察：与微颗粒共培养的巨噬细胞是否被染成棕黄色；3.电镜观察：与微颗粒共培养后的巨噬细胞其核内是否含有HPV病毒颗粒；4.双光子荧光显微镜观察：与微颗粒共培养后，，巨噬细胞可否吞噬尖锐湿疣组织来源的微颗粒及微颗粒中是否含有HPV病毒颗粒，巨噬细胞吞噬微颗粒后是否与溶酶体结合。结果：1. RT-PCR方法检测结果：每次（共四次）巨噬细胞与微颗粒共培养后，其基因组中HPV6/11-DNA表达均为阳性。2.免疫细胞化学方法观察：与微颗粒共培养的巨噬细胞中HPV染色呈棕黄色阳性反应。3.电子显微镜观察：与微颗粒共培养的巨噬细胞其核中存在HPV病毒颗粒。4.双光子荧光显微镜观察：经荧光染色后，绿色的巨噬细胞中含有红色的微颗粒，红色的微颗粒中则含有绿色荧光染色的HPV；巨噬细胞中红色溶酶体与绿色微颗粒未见结合。结论：1. HPV可通过微颗粒的方式感染巨噬细胞。2.巨噬细胞感染HPV后还能产生并释放新的含HPV的微颗粒。
[Abstract]:Objective: to explore the pathway of human papillomavirus (HPV) infection in macrophages. Method 1: 1. The microparticles extracted from (CA) of condyloma acuminatum tissue and macrophages isolated from the peripheral blood of healthy people were co-cultured, the cultured macrophages were collected and the microparticles in the supernatant were extracted. The microparticles extracted from the supernatant were co-cultured with the newly isolated macrophages, and then co-cultured for four times. RT-PCR was used to detect the presence of HPV-DNA;2. in the genome of each co-cultured macrophage. Immunocytochemical observation: macrophages co-cultured with microparticles were stained brown. Electron microscope observation: the macrophages co-cultured with microparticles contained HPV virus particles in their nuclei. Two-photon fluorescence microscope observation: after co-culture with microparticles, can macrophage phagocytosis of condyloma acuminatum tissue microparticles and whether there are HPV virus particles, macrophages phagocytosis of micro-particles and lysosome binding. The result is 1: 1. The results of RT-PCR assay showed that the HPV6/11-DNA expression was positive in the genome of macrophages cultured with microparticles each time (four times). Observation of immunocytochemistry: HPV staining in macrophages co-cultured with microparticles was brown. 3. Electron microscope observation: macrophages co-cultured with microparticles have HPV virus particles. 4. 4 in their nuclei. Two-photon fluorescence microscope observation showed that green macrophages contained red microparticles in green macrophages, while red lysosomes in green fluorescent stained HPV; macrophages did not bind with green microparticles. Conclusion 1. HPV can infect macrophages by microparticles. Macrophages infected with HPV also produce and release new HPV-containing microparticles.
【学位授予单位】：华中科技大学
【学位级别】：硕士
【学位授予年份】：2012
【分类号】：R752.53

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