本文選題：MHV-3 + Balb/cJ��； 參考：《華中科技大學》2006年碩士論文

【摘要】： 目的鑒于病毒性肝炎主要為免疫介導的肝細胞炎性損傷過程,本課題旨在探索T細胞亞群在不同臨床類型小鼠病毒性肝炎發(fā)生發(fā)展過程中的作用,為病毒性肝炎的免疫治療提供新的思路。 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),采用流式細胞儀檢測感染后0、24、48、72 h外周血、脾細胞懸液以及肝臟組織中T淋巴細胞亞群包括CD3~+CD4~+CD8-、CD3~+CD4-CD8~+及CD3~+CD4-CD8-T(DN T)細胞的細胞數(shù)及各自在T淋巴細胞中所占比率。利用免疫磁珠分選出DN T細胞(效應細胞)和CD8~+ T細胞(靶細胞),然后采用MTT法(非特異性殺傷試劑盒)進行殺傷實驗。 結(jié)果Balb/cJ小鼠于感染后24h外周血、脾臟以及肝臟T淋巴細胞中DN T細胞比率明顯升高(外周血中由2.11%升至10.35%、脾臟中由9.37%升至34.38%、肝臟中由4.05%升至56.28%)直至小鼠3d后全部死亡,CD3~+CD4~+CD8-和CD3~+CD4-CD8~+T淋巴細胞比率相應下降;A/J小鼠感染后24至96h外周血和脾臟T細胞各亞群變化不明顯。殺傷實驗提示隨著效靶比的升高,DN T細胞的殺傷活性呈現(xiàn)遞增趨勢。 結(jié)論兩種不同品系小鼠感染MHV-3后的不同轉(zhuǎn)歸、DN T細胞比率的不同改變及DN T細胞對于CD8~+ T細胞的抑制作用提示DN T細胞可能在病毒性肝炎發(fā)生、發(fā)展中起到重要作用。 研究意義 HBV感染個體后可導致不同的臨床轉(zhuǎn)歸和病理類型,包括急性乙型肝炎(輕,中,重度)和暴發(fā)性肝功能衰竭;肝臟病理學改變可表現(xiàn)為從輕微的炎性細胞侵潤,不伴有明顯的肝細胞損害(輕度)至大面積肝組織壞死,大量炎性細胞侵潤,廣泛微血管血栓形成(重度或暴發(fā)型)。且HBV感染后病情易發(fā)生慢性遷延或免疫耐受-慢性肝炎(輕,中,重度)提示不同個體對HBV感染的免疫應答有著顯著差異,機體某類內(nèi)源性細胞的保護效應可能起重要作用,但機制至今不明。利用不同品系小鼠感染MHV-3后所致不同臨床類型病毒性肝炎作為研究對象,深入開展此領域的研究并建立局部保護效應的細胞克隆將有可能為從根本上解決重癥肝炎的治療問題提供理論依據(jù)。此群細胞的正確使用亦極有希望打破機體免疫耐受,為慢性肝炎治療帶來福音。
[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.
【學位授予單位】：華中科技大學
【學位級別】：碩士
【學位授予年份】：2006
【分類號】：R512.6;R392

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