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树突状细胞亚群在SIVmac239感染中国恒河猴中数量、表型和功能变化及其机制的研究
其他题名The Dynamic, Immunophenotypic and Functional Changes of Dendritic Cell Subsets in SIVmac239-Infected Chinese Rhesus Macaques
夏厚军
学位类型博士
导师郑永唐
2010
学位授予单位中国科学院研究生院
学位授予地点北京
关键词艾滋病 Hiv Sivmac239 树突状细胞 中国恒河猴 动物模型
摘要本论文主要由3 个相对独立的部分组成:中国恒河猴单核细胞来源的树突 状细胞的表型及功能研究;外周血DC 亚群在SIVmac239 感染的中国恒河猴中 数量及细胞因子的变化以及急性感染期SIVmac239 对中国恒河猴外周血DC 亚 群的凋亡和免疫表型的影响。 非人灵长类动物是人类的近亲,由于在组织结构、免疫、生理和代谢等诸 多方面与人类高度近似,科学界较普遍地利用非人灵长类作为动物模型来进行 艾滋病(AIDS)的发病机制和疫苗研究。中国恒河猴发病缓慢,更适合于HIV 感染的相关研究。在本研究中,我们在体外成功培养了中国恒河猴单核细胞来 源的树突状细胞(monocyte derived dendritic cells,MDDC),并测定其表型和免 疫刺激功能。通过GM-CSF 和IL-4 共同刺激培养单核细胞6 天以后便获得了未 成熟MDDC,随后加入IL-1β、PGE2、LPS 和TNF-α 联合刺激MDDC 成熟。 成熟的MDDC 上调了共刺激分子和CD83 的表达,具有很强的刺激T 淋巴细胞 增殖的能力并分泌大量的IL-12。本研究为后续的DC 疫苗研究奠定了基础。 我们实验室建立了SIVmac239 感染的中国恒河猴动物模型。以该模型为依 托,我们研究了外周血中DC 亚群在急性感染期以及慢性感染期的数量、表型 及功能变化。DC 作为最重要的连接先天免疫与获得性免疫的抗原递呈细胞, 在AIDS 发病进程中扮演着重要的角色。研究发现AIDS 患者血液和淋巴结中 髓样DC(myeloid DC,mDC)和浆细胞样DC(plasmacytoid DC,pDC)会随 着感染的进程而减少,并且伴随着功能损伤。本论文通过研究发现,中国恒河 猴的DC 亚群数量在感染后尽管波动十分剧烈,但并没有显著性地增加或减少, 中文摘要 2 在后期DC 数量能够回升到正常的范围之类,这种回升不同于印度恒河猴,很 可能是中国恒河猴缓慢发病的原因之一。进一步通过研究体外刺激DC 亚群分 泌的细胞因子,我们发现在急性感染期,pDC 分泌的IFN-α 显著提高,并很可 能刺激mDC 成熟并促进了IL-12 的分泌。早期大量细胞因子的分泌有助于控制 病毒复制,但同时也激起了整个免疫系统的活化,促进了疾病进程。而在整个 感染阶段,IFN-α 与CD4+ T 细胞呈正相关,而与病毒载量呈负相关,表明了 IFN-α 对于延缓疾病进程具有重要的意义。 我们测定了急性感染期DC 亚群受病毒影响而发生的表型变化,发现pDC 更容易受到病毒影响而发生凋亡,这可能与pDC 高表达SIV 受体CD4 和CCR5 有关。在感染过程中,尽管mDC 和pDC 都显著地下调了CD4 表达,而上调了 CCR5 的表达,不过仅发现pDC CD4 的表达与病毒载量呈负相关,而CCR5 的 表达与病毒载量呈正相关。在此过程中DC 亚群都会因为病毒的影响而活化, 继而提高CCR7 的表达。同时无论mDC 还是pDC,其表达的CD80 和CD86 都与病毒载量呈正相关。在早期感染中,DC 的活化促使整个免疫系统针对病 毒发挥免疫反应,对于控制疾病发展具有重要意义。
其他摘要This dissertation was composed of three correspondingly independent parts. Phenotype and function of monocytes-derived dendritic cells (MDDC) from Chinese rhesus macaques, dynamic and functional changes of blood myeloid and plasmacytoid dendritic cells during SIVmac239 infection in Chinese Rhesus macaques and effect of SIVmac239 infection on apoptosis and immunophenotype of blood CD1c+ myeloid and plasmacytoid dendritic cells in Chinese Rhesus Macaques. Non-human primates are abroad used in AIDS researches as animal models for preclinical study of potential therapeutic drugs, vaccines and mechanisms. Compared with Indian rhesus macaques, SIV infected Chinese rhesus macaques are more suitable for AIDS researches. We first cultured and characterized MDDC from Chinese rhesus macaques. Monocytes were cultured in RPMI-1640 with GM-CSF and IL-4 for 6 days and stimulated to mature with a cocktail of IL-1β, PGE2, LPS and TNF-α. Mature MDDC increased expression of co-stimulatory molecules and CD83, and acquired a strong ability of making the allogeneic T cell proliferation and producing high level of IL-12. This study is a part work of DC vaccine research. Our lab has established the SIVmac239 infected Chinese rhesus macaques’ animal models. Depended on this model, we detected the dynamic, 英文摘要 4 immunophenotypic and functional changes of blood DC subsets. DC played a crucial role in inducing and regulating anti-HIV immune responses. Defects in the number and functions of both myeloid DC and plasmacytoid DC have been observed in the course of HIV infection and during disease progression. In our study, the number of both mDC and pDC strongly fluctuated but no significantly change during acute and chronic phases of SIVmac239 infection. The return of DC number in chronic phases may lead to a slow progression. Both the concentration of poly (I:C) induced IL-12 and HSV-1 induced IFN-α significantly increased in the acute phase of infection while returned to a normal levels at the chronic phase of infection. The peak of IFN-α emergence was earlier than that of IL-12, and it had a significantly positive correlation with IL-12, which indicated IFN-α may initiate the immune activation. We also found only the concentration of IFN-α positively correlated with CD4+ T cells counts, while negatively correlated with viral load. Our findings indicate that high levels of IFN-α in early stage of infection may contribute to the effective control of virus replication and the normal level of IFN-α in chronic infection may help Chinese rhesus macaques resist the disease progression. Followed, we studied the influence of SIV on the status of DC subsets during acute phases of infection. We found the pDC were more prone to apoptosis after infection which may be due to their high expressions of CD4 and CCR5. Both mDC and pDC decreased CD4 expression while enhanced CCR5 expression. This trend of pDC was more obviously because the mDC were low expressions of CD4 and CCR5. Interestingly, the plasma viral load was negatively correlated with CD4 MFI of pDC while positively correlated with CCR5 MFI of pDC. During this period, both mDC and pDC were active to enhance expressions of co-stimulatory molecules, accompanied with increase of CCR7. Either CD80 or CD86 expressed on mDC and pDC was positively correlated with plasma viral load. The activation of DC is benefitial to control the replication of SIV and to resist the disease progression.
语种中文
文献类型学位论文
条目标识符http://ir.kiz.ac.cn/handle/152453/6493
专题科研部门_分子免疫药理学(郑永唐)
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夏厚军. 树突状细胞亚群在SIVmac239感染中国恒河猴中数量、表型和功能变化及其机制的研究[D]. 北京. 中国科学院研究生院,2010.
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