| 题名 | HIV/HSIV 在北平顶猴PBMC 中的复制特性及APOBEC 3G/3F mRNA 的表达 |
| 作者 | 雷爱华 |
| 学位类别 | 硕士 |
| 答辩日期 | 2013-05 |
| 授予单位 | 中国科学院研究生院 |
| 授予地点 | 北京 |
| 导师 | 郑永唐 |
| 关键词 | HIV HSIV 北平顶猴 PBMC A3G A3F |
| 其他题名 | Replication characteristics of HIV/HSIV in northern pig-tailed macaque (Macaca leonina) PBMCs and APOBEC 3G/3F mRNA expression |
| 学位专业 | 生物化学与分子生物学 |
| 中文摘要 | 构建接近人艾滋病的动物模型一直是HIV/AIDS研究中的热点和难点。平顶猴细胞不表达限制HIV-1复制的限制因子TRIM5α蛋白，是目前唯一可以被HIV-1感染的旧大陆猴。平顶猴被分为3个种: 巽他平顶猴(Macaca nemestrina)、北平顶猴(Macaca leonina)和明打威猴(Macaca pagensis)。迄今为止，SIV/HIV感染平顶猴模型的研究中所使用的平顶猴基本上为巽他平顶猴。同时，研究发现限制因子APOBEC3G/3F(A3G/A3F)和HIV/SIV Vif蛋白的相互作用具有物种特异性，是限制HIV-1跨物种传播的重要屏障。最近几年，研究者发现以HIV-1为骨架，将其vif基因替换成SIVmac或SIVmne的vif基因得到的HSIV病毒能够持续感染巽他平顶猴。同时，大量研究表明HIV-1感染者中A3G和A3F mRNA的表达与患者的病程相关。 在本论文中，我们研究了替换vif基因的HIV-1病毒HSIV-vifmac、stHIV-1SV及HSIV-9N14在北平顶猴PBMC中的复制特性及感染过程中A3G和A3F mRNA的时间动态变化。首先，我们发现HIV/HSIV感染北平顶猴PBMC的能力依次为：stHIV-1SV > HSIV-vifmac > HSIV-9N14 > HIV-1NL4-3。stHIV-1SV与HSIV-vifmac能有效地感染北平顶猴PBMC，提示它们的Vif蛋白有效地拮抗了北平顶猴细胞中APOBEC3的限制作用，因而stHIV-1SV和HSIV-vifmac可用于建立北平顶猴AIDS动物模型。 此外，我们发现在北平顶猴PBMC中，A3G mRNA的表达水平与A3F mRNA的表达水平相当，有些猴中甚至低于A3F的表达水平，这与在人PBMC中的结果相反。在中国猕猴细胞中，A3G相对A3F mRNA的表达水平存在个体差异性。在HIV/HSIV感染人PBMC过程中，A3G和A3F的相对mRNA表达水平上调，且在病毒感染第6 d时平均相对mRNA的表达水平上调3倍左右。在HIV/HSIV感染北平顶猴PBMC的过程中，A3G和A3F的相对mRNA表达水平变化趋势一致，但具有较大的个体差异性。总体上，在HIV、SIV和HSIV感染北平顶猴PBMC早期(1 d ~3 d)，细胞中A3G和A3F的相对mRNA表达水平上调，且stHIV-1SV和SIVmac239感染PBMC中的A3G和A3F mRNA表达水平上调幅度较大。与HIV/HSIV感染人PBMC的结果不同，HIV/HSIV在感染北平顶猴PBMC第6 d时，A3G和A3F mRNA的表达变化不大。此外，HIV/HSIV感染人和北平顶猴PBMC过程中，A3G和A3F mRNA的相对表达水平与p24抗原浓度之间无相关性。本研究结果提示A3G和A3F的表达变化可能会影响HSIV在北平顶猴体内的复制，为建立新型的HSIV感染北平顶猴模型奠定了基础。 |
| 英文摘要 | Developing an animal model that is close to human AIDS is always an important topic in HIV/AIDS research. Notably, the pig-tailed macaque cells do not express the TRIM5α protein restricting HIV-1 replication, and pig-tailed macaque is the only primate in Old World monkeys that can be infected by HIV-1. Pig-tailed macaques have been divided into three species: sunda pig-tailed macaques (Macaca nemestrina)，northern pig-tailed macaques (Macaca leonina) and mentawai macaques (Macaca pagensis). So far, almost all the pig-tailed macaques infected by SIV/HIV are Macaca nemestrina. Studies have shown that the interaction between restriction factors APOBEC3G/3F (A3G/A3F) and HIV/SIV-derived Vif proteins is species-specificity, indicating that A3G and A3F are key barriers for cross-species transmission of HIV-1. In recent years, it has been found that substitution of vif with that from SIVmac or SIVmne enabled persistent replication of HSIV in Macaca nemestrina. Meanwhile, a number of studies have suggested that there is a correlation between A3G and A3F mRNA expression levels in HIV-1-infected patients and their disease progression. In this dissertation, we investigated the replication abilities of vif-substituted HIV-1 (HSIV-vifmac, stHIV-1SV and HSIV-9N14) in northern pig-tailed macaque PBMCs and examined the temporal dynamic changes of A3G and A3F mRNA expression during infection. We found that the ability for HIV/HSIV to infect northern pig-tailed macaque PBMCs can be listed as follows: stHIV-1SV > HSIV-vifmac > HSIV-9N14 > HIV-1NL4-3. stHIV-1SV and HSIV-vifmac can replicate robustly in PBMCs from Macaca leonina, implying that their Vif protein has overcome the restriction of APOBEC3 effectively in macaque cells. Therefore, stHIV-1SV and HSIV-vifmac can be applied to the establishment of northern pig-tailed macaque AIDS model. In addition, we found that the expression level of A3G mRNA in PBMCs of Macaca leonina is either comparable to the A3F mRNA level or even lower than A3F mRNA expression, which is contrary to the A3G/A3F mRNA expression in human PBMCs. However, the expression levels of A3G relative to A3F mRNA levels in PBMCs from Chinese rhesus macaques are donor-dependent. The A3G and A3F relative mRNA expression levels in human PBMCs can be induced by HIV/HSIV infection. Notably, the average mRNA expression levels for both A3G and A3F in HIV/HSIV-infected human PBMCs increased about 3 times higher than uninfected PBMCs at day 6. The temporal dynamic changes in A3G mRNA expression levels and in A3F mRNA levels in HIV/HSIV-infected northern pig-tailed macaque PBMCs were consistent; however, there existed large individual variations. On the whole, A3G and A3F mRNA expression levels were upregulated in PBMCs from Macaca leonina infected by HIV, SIV and HSIV at early stage (day 1 to day 3); the A3G and A3F mRNA expression levels in stHIV-1SV and SIVmac-infected macaque PBMCs were upregulated more than that in HIV-1 and HSIV-vifmac infected macaque PBMCs. In contrast to the results in HIV/HSIV-infected human PBMCs, A3G and A3F relative mRNA expression levels changed little in HIV/HSIV-infected macaque PBMCs at day 6. Additionally, there is no correlation between p24 antigen concentrations and A3G or A3F mRNA relative expression levels during HIV/HSIV infection in PBMCs from both humans and Macaca leonina. In conclusion, our results suggest that the expression changes of A3G and A3F may influence HSIV infection of northern pig-tailed macaques and has laid a foundation for establishing novel northern pig-tailed macaque models of HSIV infection in vivo. |
| 语种 | 中文 |
| 公开日期 | 2013-06-17 |
| 内容类型 | 学位论文 |
| 源URL | [http://159.226.149.42:8088/handle/152453/7484]  |
| 专题 | 昆明动物研究所_分子免疫药理学 |
| 推荐引用方式 GB/T 7714 | 雷爱华. HIV/HSIV 在北平顶猴PBMC 中的复制特性及APOBEC 3G/3F mRNA 的表达[D]. 北京. 中国科学院研究生院. 2013. |
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