Bcl11a和Bcl11b基因抑制病毒增殖功能研究
发布时间：2018-07-20 11:46
【摘要】：Bcl11a和Bcl11b是锌指蛋白样转录因子,分别对B淋巴和T淋巴细胞的发育、分化和增殖起到重要调节作用。已有的研究也表明Bcl11a和Bcl11b作为核转录因子调控了一系列下游基因的表达,对细胞的增殖、周期调控和免疫的调节发挥了重要作用。同时,这两个基因还被证实不仅与癌症有密切关系,也可以抑制HIV-1病毒在细胞内的转录和复制,发挥一定的抗病性。鉴于Bcl11a和Bcl11b基因在淋巴细胞中的特异表达和对淋巴细胞生长发育具有重要作用,然而,在免疫反应及抗猪圆环病毒PCV2的功能不清楚,本研究以小鼠和PK15细胞为试验对象,猪圆环病毒2型PCV2为病原,主要采用基因敲除、基因超表达、荧光定量、Western blot和免疫荧光等方法,对Bcl11a和Bcl11b抑制PCV2增殖的作用和机理进行了初步研究。同时,还以昆明小鼠为攻毒对象,初步探讨了PCV2对淋巴细胞发育的影响。主要研究成果如下:(1)半敲除Bcl11a后(Bcl11a+/-,Bcl11a表达量降低一半)小鼠免疫能力显著降低,血清中PCV2病毒含量显著上升,PCV2对脾脏细胞的感染能力也增强,对脾脏等免疫器官的损伤更加严重。(2)超表达Bcl11a和Bcl11b能显著抑制PCV2在PK15细胞中的增殖,病毒m RNA、DNA和蛋白水平都显著降低。(3)Bcl11a、Bcl11b和PCV2病毒没有明显的共定位。(4)Bcl11a可以通过抑制Caspase3的活性抑制由PCV2引起的细胞凋亡,而Bcl11b通过抑制NF-κB的表达并促进抗凋亡蛋白bcl2、抗病毒蛋白IRF3和MX1的表达,抑制病毒增殖,减少细胞凋亡,说明Bcl11a、Bcl11b抑制病毒增殖的机制可能是通过不同途径实现的。(5)PCV2能导致发育中的淋巴细胞大量损耗,尤其是对B淋巴细胞的影响非常明显。PCV2也可以抑制体外培养淋巴细胞的增殖,并对淋巴细胞发育相关重要基因有显著的抑制作用,说明PCV2可以抑制早期淋巴细胞的发育和增殖。
[Abstract]:Bcl11a and Bcl11b are zinc finger protein like transcription factors, which play an important role in regulating the development, differentiation and proliferation of B lymphatic and T lymphocytes. The previous studies have also shown that Bcl11a and Bcl11b have regulated the expression of a series of downstream genes as nuclear transcription factors, which play an important role in cell proliferation, cycle regulation and immune regulation. At the same time, these two genes have also been proved not only closely related to cancer, but also inhibit the transcription and replication of HIV-1 virus in cells and play a certain resistance to disease. In view of the specific expression of Bcl11a and Bcl11b genes in lymphocytes and the important role in lymphocyte growth and development, however, in the immune response and anti porcine circovirus. The function of PCV2 is not clear. In this study, mice and PK15 cells were used as the test object. The porcine circovirus type 2 PCV2 was the pathogen. The effect and mechanism of Bcl11a and Bcl11b on the inhibition of PCV2 proliferation were primarily studied by gene knockout, gene overexpression, fluorescence quantitative, Western blot and immunofluorescence. Meanwhile, the mice were attacked by Kunming mice. The effect of PCV2 on the development of lymphocyte was preliminarily discussed. The main research results were as follows: (1) after half knockout Bcl11a (Bcl11a+/-, Bcl11a expression was reduced by half), the immune ability of mice decreased significantly, the content of PCV2 virus in serum increased significantly, the infection ability of PCV2 to spleen cells increased, and the spleen and other immune organs were more damaged. (2) overexpression of Bcl11a and Bcl11b could significantly inhibit the proliferation of PCV2 in PK15 cells, and the level of M RNA, DNA and protein decreased significantly. (3) Bcl11a, Bcl11b and PCV2 viruses have no obvious co location. (4) Bcl11a can inhibit the apoptosis induced by the inhibition of Caspase3 activity. Promoting the expression of anti apoptotic protein BCL2, antiviral protein IRF3 and MX1, inhibiting the proliferation of virus and reducing cell apoptosis, the mechanism of Bcl11a, Bcl11b to inhibit the proliferation of the virus may be realized through different ways. (5) PCV2 can lead to a large amount of lymphocyte depletion in developing cells, especially the effect on B lymphocytes very obviously.PCV2 can also be suppressed. The proliferation of lymphocytes was cultured in vitro, and it has a significant inhibitory effect on the key genes related to lymphocyte development. It shows that PCV2 can inhibit the development and proliferation of early lymphocyte.
【学位授予单位】：华中农业大学
【学位级别】：硕士
【学位授予年份】：2015
【分类号】：S852.65
本文编号：2133414
[Abstract]:Bcl11a and Bcl11b are zinc finger protein like transcription factors, which play an important role in regulating the development, differentiation and proliferation of B lymphatic and T lymphocytes. The previous studies have also shown that Bcl11a and Bcl11b have regulated the expression of a series of downstream genes as nuclear transcription factors, which play an important role in cell proliferation, cycle regulation and immune regulation. At the same time, these two genes have also been proved not only closely related to cancer, but also inhibit the transcription and replication of HIV-1 virus in cells and play a certain resistance to disease. In view of the specific expression of Bcl11a and Bcl11b genes in lymphocytes and the important role in lymphocyte growth and development, however, in the immune response and anti porcine circovirus. The function of PCV2 is not clear. In this study, mice and PK15 cells were used as the test object. The porcine circovirus type 2 PCV2 was the pathogen. The effect and mechanism of Bcl11a and Bcl11b on the inhibition of PCV2 proliferation were primarily studied by gene knockout, gene overexpression, fluorescence quantitative, Western blot and immunofluorescence. Meanwhile, the mice were attacked by Kunming mice. The effect of PCV2 on the development of lymphocyte was preliminarily discussed. The main research results were as follows: (1) after half knockout Bcl11a (Bcl11a+/-, Bcl11a expression was reduced by half), the immune ability of mice decreased significantly, the content of PCV2 virus in serum increased significantly, the infection ability of PCV2 to spleen cells increased, and the spleen and other immune organs were more damaged. (2) overexpression of Bcl11a and Bcl11b could significantly inhibit the proliferation of PCV2 in PK15 cells, and the level of M RNA, DNA and protein decreased significantly. (3) Bcl11a, Bcl11b and PCV2 viruses have no obvious co location. (4) Bcl11a can inhibit the apoptosis induced by the inhibition of Caspase3 activity. Promoting the expression of anti apoptotic protein BCL2, antiviral protein IRF3 and MX1, inhibiting the proliferation of virus and reducing cell apoptosis, the mechanism of Bcl11a, Bcl11b to inhibit the proliferation of the virus may be realized through different ways. (5) PCV2 can lead to a large amount of lymphocyte depletion in developing cells, especially the effect on B lymphocytes very obviously.PCV2 can also be suppressed. The proliferation of lymphocytes was cultured in vitro, and it has a significant inhibitory effect on the key genes related to lymphocyte development. It shows that PCV2 can inhibit the development and proliferation of early lymphocyte.
【学位授予单位】：华中农业大学
【学位级别】：硕士
【学位授予年份】：2015
【分类号】：S852.65
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