人巨噬细胞感染人乳头瘤病毒的实验研究
发布时间：2018-11-11 09:19
【摘要】：目的:探讨巨噬细胞能否被人乳头瘤病毒感染。方法:我们取门诊病人的尖锐湿疣组织制备成含HPV的混悬液,以及取健康人的外周血制备巨噬细胞,将两者混合培养,通过RT-PCR检测巨噬细胞基因组中是否有HPV-DNA;电镜观察上述培养的巨噬细胞胞核内是否含有HPV颗粒;免疫组化方法观察在尖锐湿疣组织真皮乳头层内是否有CD68阳性细胞以及含HPV颗粒的阳性细胞;免疫荧光方法观察真皮乳头内是否有对抗CD68荧光抗体和抗HPV荧光抗体均显色的同一细胞。结果:1. RT-PCR方法检测:含HPV溶液和巨噬细胞共培养,四次传代的后,巨噬细胞的基因组中均检测到HPV6/11-DNA。2.电镜观察:上述传代培养的巨噬细胞胞核中可见到HPV颗粒。3.免疫组化方法观察:在尖锐湿疣组织真皮乳头内既有CD68阳性细胞,又有含HPV的阳性细胞。4.免疫荧光观察:在尖锐湿疣组织真皮乳头内有显示HPV的绿色荧光染,显示CD68阳性细胞的橘色荧光,两张图片重叠后显示橙黄色荧光。结论:尖锐湿疣组织内的巨噬细胞能被HPV感染,这种被HPV感染的巨噬细胞的抗病毒功能很有可能被削弱。
[Abstract]:Objective: to investigate whether macrophages can be infected by human papillomavirus (HPV). Methods: we prepared suspension containing HPV from condyloma acuminatum tissue of outpatients and macrophages from peripheral blood of healthy people. The macrophages were mixed and cultured. RT-PCR was used to detect whether there was HPV-DNA; in the genome of macrophages. The HPV granules were observed by electron microscope in the nucleus of macrophages and whether there were CD68 positive cells and HPV positive cells in the dermal papillary layer of condyloma acuminatum tissue by immunohistochemical method. Immunofluorescence method was used to observe whether there were the same cells in dermal papilla that had both anti CD68 fluorescent antibody and anti HPV fluorescent antibody. The result is 1: 1. RT-PCR detection: HPV6/11-DNA.2. was detected in the genome of macrophages after four passages with HPV solution and macrophage co-cultured. Electron microscope observation: the above passage of macrophages can be seen in the nucleus of HPV particles. Immunohistochemical observation: there were CD68 positive cells and HPV positive cells in the dermal papilla of condyloma acuminatum. 4. 4. Immunofluorescence observation: in the dermal papilla of condyloma acuminatum there were green fluorescent staining of HPV and orange fluorescence of CD68 positive cells. Conclusion: macrophages in condyloma acuminatum tissue can be infected with HPV, and the antiviral function of macrophages infected with HPV is likely to be weakened.
【学位授予单位】：华中科技大学
【学位级别】：硕士
【学位授予年份】：2011
【分类号】：R752.53
本文编号：2324414
[Abstract]:Objective: to investigate whether macrophages can be infected by human papillomavirus (HPV). Methods: we prepared suspension containing HPV from condyloma acuminatum tissue of outpatients and macrophages from peripheral blood of healthy people. The macrophages were mixed and cultured. RT-PCR was used to detect whether there was HPV-DNA; in the genome of macrophages. The HPV granules were observed by electron microscope in the nucleus of macrophages and whether there were CD68 positive cells and HPV positive cells in the dermal papillary layer of condyloma acuminatum tissue by immunohistochemical method. Immunofluorescence method was used to observe whether there were the same cells in dermal papilla that had both anti CD68 fluorescent antibody and anti HPV fluorescent antibody. The result is 1: 1. RT-PCR detection: HPV6/11-DNA.2. was detected in the genome of macrophages after four passages with HPV solution and macrophage co-cultured. Electron microscope observation: the above passage of macrophages can be seen in the nucleus of HPV particles. Immunohistochemical observation: there were CD68 positive cells and HPV positive cells in the dermal papilla of condyloma acuminatum. 4. 4. Immunofluorescence observation: in the dermal papilla of condyloma acuminatum there were green fluorescent staining of HPV and orange fluorescence of CD68 positive cells. Conclusion: macrophages in condyloma acuminatum tissue can be infected with HPV, and the antiviral function of macrophages infected with HPV is likely to be weakened.
【学位授予单位】：华中科技大学
【学位级别】：硕士
【学位授予年份】：2011
【分类号】：R752.53
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