本文選題：口蹄疫病毒 + 滅活疫苗�。� 參考：《中國農(nóng)業(yè)科學(xué)院》2016年碩士論文

【摘要】：有研究發(fā)現(xiàn)動物免疫口蹄疫滅活疫苗后,機(jī)體內(nèi)抗口蹄疫病毒抗體水平與攻毒后動物的保護(hù)力之間存在密切聯(lián)系。根據(jù)攻毒前機(jī)體內(nèi)抗體水平與攻毒后動物保護(hù)力之間的關(guān)系可將抗口蹄疫病毒抗體水平進(jìn)行分類,分別為:抗體白色區(qū),即完全保護(hù)區(qū),體內(nèi)抗體水平很高,動物感染口蹄疫病毒后處于完全保護(hù)狀態(tài);黑色區(qū),即完全不保護(hù)區(qū),體內(nèi)抗體水平很低,動物感染口蹄疫病毒后全部發(fā)病;灰色區(qū),即部分保護(hù)區(qū),體內(nèi)抗體水平處于白色區(qū)和黑色區(qū)之間,動物感染口蹄疫病毒后,有些動物處于保護(hù)狀態(tài),而有些動物會發(fā)病。在灰色區(qū)內(nèi)還會出現(xiàn)抗體水平高的動物會發(fā)病,而抗體水平較低的動物得到保護(hù)的現(xiàn)象。本研究的主要目的是探究抗體灰色區(qū)內(nèi)動物保護(hù)力不同的機(jī)制。為探索口蹄疫免疫抗體灰色區(qū)形成的機(jī)制,本實驗選取33頭健康長白豬免疫自行制備的O型口蹄疫滅活疫苗,于免疫后28天進(jìn)行同源強(qiáng)毒攻毒,隨后連續(xù)觀察十天確定動物發(fā)病情況。于免疫前及免疫后第28天采集血液樣品進(jìn)行試驗,使用液相阻斷ELISA測定血清中抗體水平,流式細(xì)胞術(shù)測定外周血中淋巴細(xì)胞亞群含量,抗體芯片技術(shù)測定血清中細(xì)胞因子含量。根據(jù)免疫后第28天時血清中抗體水平與攻毒后動物發(fā)病情況的關(guān)系,確定免疫抗體灰色區(qū)的范圍。結(jié)果表明豬免疫口蹄疫滅活疫苗后,灰色區(qū)抗體效價的范圍為1:6-1:45,灰色區(qū)動物在免疫前后CTL細(xì)胞含量及TNF-α、IFN-γ、IL-12、IL-15、IL-18、MIG、GM-CSF、IL-1α、TGF-α、TWEAK-R等細(xì)胞因子的含量均發(fā)生變化�；疑珔^(qū)內(nèi)保護(hù)組動物免疫后CTL細(xì)胞含量與免疫前相比出現(xiàn)升高,而發(fā)病動物則出現(xiàn)顯著性降低。對于IFN-γ、IL-12、GM-CSF、IL-1α、IL-15和TNF-α這幾種細(xì)胞因子,其含量在灰色區(qū)保護(hù)組動物中免疫后與免疫前相比出現(xiàn)升高,而發(fā)病動物則沒有發(fā)生變化或出現(xiàn)不同程度的降低。MIG和TWEAK R在保護(hù)組動物中免疫前后沒有明顯變化,但是發(fā)病動物其含量出現(xiàn)下降。而對于IL-18和TGF-α,保護(hù)組動物在免疫后其含量低于免疫前,不保護(hù)組動物IL-18的含量在免疫后反而出現(xiàn)上升,TGF-α的含量沒有明顯變化。實驗結(jié)果顯示,負(fù)責(zé)細(xì)胞免疫應(yīng)答的CTL細(xì)胞及TNF-α、IFN-γ、IL-12和IL-15等Th1型細(xì)胞因子能夠提高灰色區(qū)內(nèi)動物抗口蹄疫病毒感染的能力,由此可得出免疫后誘導(dǎo)的細(xì)胞免疫應(yīng)答強(qiáng)弱與口蹄疫抗體灰色區(qū)內(nèi)動物保護(hù)力的高低有關(guān)。同時選用另外33頭健康長白豬免疫口蹄疫滅活疫苗,觀察整個免疫階段外周血淋巴細(xì)胞亞群動態(tài)變化。分別于免疫前,免疫后第1、3、5、7、10、14、21、35天采集抗凝血,進(jìn)行流式細(xì)胞術(shù)實驗檢測血液中細(xì)胞亞群的分布情況,同時測定血清中抗體水平。結(jié)果顯示,在整個免疫期Th細(xì)胞含量變化與B細(xì)胞含量變化存在相關(guān)性,同時也與抗體水平的變化相關(guān),這表明了Th細(xì)胞與B細(xì)胞之間存在相互作用的關(guān)系,共同介導(dǎo)體液免疫應(yīng)答。DCs含量在免疫后快速升高,說明其在短期內(nèi)即可被活化發(fā)揮功能。結(jié)果表明,疫苗免疫后DCs能快速發(fā)揮功能,使固有免疫應(yīng)答朝著適應(yīng)性免疫應(yīng)答的方向發(fā)展。B細(xì)胞和Th細(xì)胞含量的升高說明體液免疫應(yīng)答被激活,產(chǎn)生的高水平和高度特異性的抗體可對機(jī)體起到保護(hù)作用,同時CTL細(xì)胞含量也出現(xiàn)升高,說明細(xì)胞免疫應(yīng)答在該過程中也發(fā)揮了至關(guān)重要的作用。
[Abstract]:There is a close relationship between the level of anti FMDV antibody and the protection ability of the animals after inoculation of the inactivated vaccine against FMDV. The anti body level of FMD virus can be classified according to the relationship between the level of antibody in the body of the virus and the protective ability of the animal after the attack, which is the white area of the antibody, respectively. In the complete protection area, the level of antibody in the body is very high, the animal infected with FMDV is completely protected, the black area, that is, the completely non protected area, the level of antibody in the body is very low, the animal infected with the foot and mouth disease virus, and the grey area, that is, part of the protective area, the body anti body level is between the white area and the black area, and the animal infect the mouth of the mouth. After pestilence virus, some animals are in protective condition, and some animals will occur. In the gray area, there will be a high level of antibody in animals, and the animals with low antibody level are protected. The main purpose of this study is to explore the mechanism of different animal protection ability in the gray area of the antibody. The mechanism of color region formation was selected by 33 healthy long white pigs immunized with O inactivated vaccine of foot and mouth disease (FMDV). After 28 days of immunization, it was observed for ten days for ten days. The blood samples were collected before and twenty-eighth days after immunization, and the serum resistance was measured by liquid phase blocking ELISA. The content of lymphocyte subsets in peripheral blood was measured by flow cytometry, and the content of cytokines in serum was determined by antibody chip technique. The range of grey area of immune antibody was determined according to the relationship between the serum antibody level and the incidence of attack after twenty-eighth days after immunization. The range of antibody titer was 1:6-1:45, and the content of CTL cells in the gray area and the content of TNF- alpha, IFN- gamma, IL-12, IL-15, IL-18, MIG, GM-CSF, IL-1 a, TGF- alpha, TWEAK-R and other cytokines were all changed. For IFN- gamma, IL-12, GM-CSF, IL-1 a, IL-15 and TNF- alpha, the content of these cytokines increased in the grey area protection group after immunization, but the incidence of.MIG and TWEAK R in the animals did not change significantly, but there was no obvious change in the immune system in the animals of the protective group. The content of the sick animals decreased. For IL-18 and TGF- alpha, the contents of the animals in the protection group were lower than those before the immune system. The content of IL-18 in the non protective group rose but the content of TGF- alpha did not change obviously. The experimental results showed that the CTL cells and TNF- alpha, IFN- gamma, IL-12 and IL-15, which were responsible for the cellular immune response. Cytokines can improve the ability of animals to resist foot and mouth disease virus infection in the gray area, thus it can be concluded that the immune response induced by immune response is related to the level of animal protection in the grey area of FMD antibody. At the same time, 33 other healthy long white pigs immunized foot-and-mouth vaccine was selected to observe the peripheral blood lymph nodes in the whole immune phase. Dynamic changes in cell subgroups. Anticoagulants were collected on day 1,3,5,7,10,14,21,35 before immunization, and the distribution of cell subgroups in blood was detected by flow cytometry and serum antibody levels were measured at the same time by flow cytometry. The results showed that there was a correlation between the changes of Th cells in the whole immune period and the changes of B cells in the whole immune period, and also in the same time. The change of antibody level is associated with the interaction between Th cells and B cells. The.DCs content of the common dielectric liquid immune response is rapidly increased after immunization, indicating that it can be activated in the short term. The result shows that the immune response of the vaccine to the immune response is fast and that the intrinsic immune response will be adapted to the adaptive immune response. The increase in the content of.B and Th cells indicates that the humoral immune response is activated. The high level and highly specific antibodies can protect the body, and the content of CTL cells also increases, indicating that the cellular immune response also plays a vital role in this process.
【學(xué)位授予單位】：中國農(nóng)業(yè)科學(xué)院
【學(xué)位級別】：碩士
【學(xué)位授予年份】：2016
【分類號】：S858.28

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本文編號：2071988