本文選題：MHV-3　切入點：C3H/HeJ　出處：《華中科技大學(xué)》2006年碩士論文

【摘要】： 目的: 慢性病毒性肝炎是一種嚴(yán)重危害人類健康的常見病,多發(fā)病,目前尚缺乏理想的實驗動物模型來闡明其具體的發(fā)病機制,從而使得對其防治的研究受到很大的局限。本課題擬克服現(xiàn)有模型所存在的不足,建立一種更為理想的慢性病毒性肝炎實驗動物模型,為慢性病毒性肝炎發(fā)病機制的研究以及防治提供新的理論依據(jù)。在此模型基礎(chǔ)上我們對其外周血及脾臟T細胞亞群的動態(tài)變化做初步觀察,以探討病毒性肝炎慢性遷延免疫機制,并為其免疫治療提供一些新的思路。 方法: 取6~-8周齡雌性C3H/HeJ小鼠60只,將純化MHV-3病毒進行稀釋,以10pfu/200ul/只行C3H/HeJ小鼠腹腔注射,然后對其一般情況、生存曲線進行觀察,并于病毒感染后4、6、8、9、10、12、15、20、25、30及40天各取C3H小鼠三只行肝臟組織病理學(xué)、血液生化指標(biāo)以及肝臟組織病毒滴度的變化進行檢測。同時采用流式細胞儀檢測感染后相應(yīng)時間點外周血、脾細胞懸液中T淋巴細胞亞群包括CD3~+CD4~+CD8~-、CD3~+CD4~-CD8~+、CD3~+CD4~+CD25~+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。以生理鹽水注射組小鼠作為對照。 結(jié)果: C3H/HeJ小鼠感染MHV-3后,6天左右開始出現(xiàn)少部分小鼠死亡,約63%存活,存活下來的小鼠一般情況較正常對照組差,肝臟組織呈現(xiàn)持續(xù)炎性改變,血清ALT、AST升高而TP、ALB有所下降,且肝臟組織病毒滴度增高,呈現(xiàn)持續(xù)病毒復(fù)制,與人類慢性病毒性肝炎的發(fā)生發(fā)展極為類似。其外周血中及脾臟中CD3~+CD4~+CD25~+及CD3~+CD4~-CD8~-T(DN T)細胞在T細胞中的比率有升高的趨勢。 結(jié)論: 本研究所建立的實驗動物模型與人類慢性病毒性肝炎的發(fā)生發(fā)展極為類似,在目前尚無更好的模型的前提下,不失為一種較為理想的慢性病毒性肝炎實驗動物模型。其外周血及脾臟中CD3~+CD4~+CD25~+及CD3~+CD4~-CD8~-T(DN T)細胞比率的升高趨勢提示這兩類細胞可能在慢性病毒性肝炎發(fā)生、發(fā)展中起著一定的作用。本研究的意義: 本實驗所選鼠肝炎3型病毒(MHV-3)易于體內(nèi)、體外培養(yǎng), C3H/HeJ小鼠遺傳背景清晰,所建模型成本相對低廉,重復(fù)性好,同時也克服了已有慢性病毒性肝炎動物模型體內(nèi)缺乏病原體復(fù)制而與臨床情況顯著不同的局限性,使系統(tǒng)性、動態(tài)性研究慢性病毒性肝炎的遺傳控制因素及分子生物學(xué)、分子免疫學(xué)發(fā)病機理等成為可能,為其防治提供新的理論依據(jù)。在目前尚無完善的慢性病毒性肝炎動物模型的情況下,不失為一種較好的慢性病毒性肝炎動物模型。目前免疫調(diào)節(jié)T細胞包括CD3~+CD4~+CD25~+及CD3~+CD4~-CD8~-T(DN T)等的研究是一個熱點,我們在此模型基礎(chǔ)上對外周血及脾臟中T細胞亞群的動態(tài)變化進行了初步觀察,發(fā)現(xiàn)在此模型中這兩群調(diào)節(jié)T細胞的比率都呈現(xiàn)上升趨勢,提示這兩類細胞可能在慢性病毒性肝炎發(fā)生、發(fā)展中起著一定的作用,為慢性病毒性肝炎中免疫調(diào)節(jié)T細胞的具體作用及機制的深入研究打下基礎(chǔ)。
[Abstract]:Objective:
Chronic viral hepatitis is a common disease seriously harming human health, disease, lack of ideal animal models to elucidate its pathogenesis, which makes the study on its prevention and treatment are very limited. This paper intends to overcome the shortcomings of the existing model, establish a more chronic virus hepatitis A animal model, provide a new theoretical basis for the study of the pathogenesis of chronic viral hepatitis prevention and control. Based on this model we dynamic changes of peripheral blood and spleen T cell subsets do preliminary observation, to explore the immune mechanism of chronic viral hepatitis, and provide some new ideas for the immune therapy.
Method:
60 female C3H/HeJ mice aged 6~-8 weeks, the purified MHV-3 virus was diluted to 10pfu/200ul/ C3H/HeJ mice by intraperitoneal injection, and then the general condition, survival curves were observed, and in 40 days after infection of 4,6,8,9,10,12,15,20,25,30 and the three C3H mice liver histopathology, blood biochemical indices and the change of liver the organization of virus titer was detected. At the same time were detected by flow cytometry after infection at corresponding time of peripheral blood, spleen cell suspension in T lymphocyte subsets including CD3~+CD4~+CD8~-, CD3~+CD4~-CD8~+, CD3~+CD4~+, CD25~+ and CD3~+CD4~-CD8~-T (DN T) the number of cells in T lymphocytes and their proportion. In the saline group mice were used as controls.
Result:
The C3H/HeJ mice infected with MHV-3 after 6 days began to appear a few mice died, about 63% survive, survive the general condition of mice compared with normal control group, the liver tissue showed persistent inflammatory changes, serum ALT, AST and TP increased, ALB decreased, and the liver tissue virus titer increased, showing sustained viral replication with human chronic viral hepatitis is very similar. The occurrence and development of CD3~+CD4~+CD25~+ and CD3~+CD4~-CD8~-T in peripheral blood and spleen (DN T) cell ratio in T cells increased.
Conclusion:
The experimental animal model with human chronic viral hepatitis established in the study of the occurrence and development of extremely similar, there is no better model under the premise, it is an ideal experimental animal model of chronic viral hepatitis. CD3~+CD4~+CD25~+ and CD3~+CD4~-CD8~-T in peripheral blood and spleen (DN T) increased the percentage of cells that these two kinds of cells may occur in patients with chronic viral hepatitis, plays a certain role in the development of the significance of this study:
The selected mouse hepatitis virus type 3 (MHV-3) to the in vivo, in vitro, C3H/HeJ mice model clear genetic background, relatively low cost, good repeatability, and overcomes the shortcomings of existing chronic viral hepatitis animal model lacking pathogen replication but significantly different from the clinical condition of the limitations of the system. The genetic factors controlling the dynamics of chronic viral hepatitis and molecular biology, molecular immunology and other possible pathogenesis, provide a new theoretical basis for its prevention and treatment. At present there is no perfect chronic viral hepatitis animal model, it is a good animal model of chronic viral hepatitis. The immune regulation of T cells including CD3~+CD4~+CD25~+ and CD3~+CD4~-CD8~-T (DN T) and so on is a hot spot, the dynamic changes of peripheral blood and spleen T cell subsets based on us A preliminary observation, found that the two group of regulatory T cell ratio showed an upward trend in this model, suggesting that these two cell types may occur in patients with chronic viral hepatitis, which plays a certain role in the development, lay the foundation for further study the specific role and mechanism of regulation of T cell immunity in chronic viral hepatitis..

【學(xué)位授予單位】：華中科技大學(xué)
【學(xué)位級別】：碩士
【學(xué)位授予年份】：2006
【分類號】：R-332

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