不同临床类型小鼠肝炎模型T细胞亚群的初步研究

发布时间：2018-05-03 01:35

本文选题：MHV-3 + Balb/cJ　；参考：《华中科技大学》2006年硕士论文

【摘要】： 目的鉴于病毒性肝炎主要为免疫介导的肝细胞炎性损伤过程,本课题旨在探索T细胞亚群在不同临床类型小鼠病毒性肝炎发生发展过程中的作用,为病毒性肝炎的免疫治疗提供新的思路。方法取Balb/cJ和A/J小鼠各8只,腹腔注射100pfu 3型鼠肝炎病毒(MHV-3),采用流式细胞仪检测感染后0、24、48、72 h外周血、脾细胞悬液以及肝脏组织中T淋巴细胞亚群包括CD3~+CD4~+CD8-、CD3~+CD4-CD8~+及CD3~+CD4-CD8-T(DN T)细胞的细胞数及各自在T淋巴细胞中所占比率。利用免疫磁珠分选出DN T细胞(效应细胞)和CD8~+ T细胞(靶细胞),然后采用MTT法(非特异性杀伤试剂盒)进行杀伤实验。结果Balb/cJ小鼠于感染后24h外周血、脾脏以及肝脏T淋巴细胞中DN T细胞比率明显升高(外周血中由2.11%升至10.35%、脾脏中由9.37%升至34.38%、肝脏中由4.05%升至56.28%)直至小鼠3d后全部死亡,CD3~+CD4~+CD8-和CD3~+CD4-CD8~+T淋巴细胞比率相应下降;A/J小鼠感染后24至96h外周血和脾脏T细胞各亚群变化不明显。杀伤实验提示随着效靶比的升高,DN T细胞的杀伤活性呈现递增趋势。结论两种不同品系小鼠感染MHV-3后的不同转归、DN T细胞比率的不同改变及DN T细胞对于CD8~+ T细胞的抑制作用提示DN T细胞可能在病毒性肝炎发生、发展中起到重要作用。研究意义 HBV感染个体后可导致不同的临床转归和病理类型,包括急性乙型肝炎(轻,中,重度)和暴发性肝功能衰竭;肝脏病理学改变可表现为从轻微的炎性细胞侵润,不伴有明显的肝细胞损害(轻度)至大面积肝组织坏死,大量炎性细胞侵润,广泛微血管血栓形成(重度或暴发型)。且HBV感染后病情易发生慢性迁延或免疫耐受-慢性肝炎(轻,中,重度)提示不同个体对HBV感染的免疫应答有着显著差异,机体某类内源性细胞的保护效应可能起重要作用,但机制至今不明。利用不同品系小鼠感染MHV-3后所致不同临床类型病毒性肝炎作为研究对象,深入开展此领域的研究并建立局部保护效应的细胞克隆将有可能为从根本上解决重症肝炎的治疗问题提供理论依据。此群细胞的正确使用亦极有希望打破机体免疫耐受,为慢性肝炎治疗带来福音。
[Abstract]:Objective to explore the role of T cell subsets in the development of viral hepatitis in mice. To provide a new idea for immunotherapy of viral hepatitis. Methods eight Balb/cJ and eight AP-J mice were injected intraperitoneally with 100pfu 3 mouse hepatitis virus (MHV-3). The peripheral blood was detected by flow cytometry for 72 hours after infection. The number of T lymphocyte subsets in spleen cell suspension and liver tissue including CD3- CD4- CD8-CD34-CD4-CD8- and CD3- CD4-CD8-T(DN T cells and their respective percentages in T lymphocytes. DN T cells (effector cells) and CD8T cells (target cells) were separated by immunomagnetic beads and then killed by MTT assay (non-specific killing kit). Results the peripheral blood of Balb/cJ mice was 24 hours after infection. The ratio of DN T cells in spleen and liver T lymphocytes increased significantly (from 2.11% to 10.35 in peripheral blood, from 9.37% to 34.38 in spleen, from 4.05% to 56.28% in liver) until all the mice died 3 days later. There was no significant change of T cell subsets in peripheral blood and spleen of AP-J mice 24 to 96 hours after infection. The killing experiments showed that the killing activity of T cells increased with the increase of the ratio of effect to target. Conclusion the different changes in the ratio of DN T cells and the inhibitory effect of DN T cells on CD8T cells in two different strains of mice infected with MHV-3 suggest that DN T cells may play an important role in the pathogenesis and development of viral hepatitis. Research significance HBV infection can lead to different clinical outcomes and pathological types, including acute hepatitis B (mild, moderate, severe) and fulminant liver failure. No obvious liver cell damage (mild) to large area liver necrosis, a large number of inflammatory cells infiltration, extensive microvascular thrombosis (severe or fulminant type). After HBV infection, chronic prolongation or immune tolerance to chronic hepatitis (mild, moderate, severe) suggest that different individuals have significant differences in immune response to HBV infection, and the protective effect of some endogenous cells may play an important role. However, the mechanism is still unknown. Different clinical types of viral hepatitis caused by MHV-3 infection in different strains of mice were used as research objects. The further research in this field and the establishment of local protective cell clones may provide a theoretical basis for the fundamental solution to the treatment of severe hepatitis. The correct use of these cells is also promising to break the immune tolerance of the body and bring good news to the treatment of chronic hepatitis.
【学位授予单位】：华中科技大学
【学位级别】：硕士
【学位授予年份】：2006
【分类号】：R512.6;R392

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