本文選題：百日咳　切入點(diǎn)：重組粘著素　出處：《昆明理工大學(xué)》2009年碩士論文

【摘要】：PRN(百日咳粘著素,Pertactin,PRN)是無(wú)細(xì)胞百日咳組分疫苗的重要組成部分。天然PRN純化工藝復(fù)雜,制備成本高,用重組PRN代替天然PRN是開(kāi)發(fā)新型百日咳疫苗的發(fā)展趨勢(shì)。本論文對(duì)重組PRN的免疫原性、抗原性和保護(hù)力進(jìn)行評(píng)價(jià),并研究其激發(fā)機(jī)體免疫應(yīng)答的類(lèi)型。 重組PRN和天然PRN交叉檢測(cè)重組PRN組和天然PRN組兩次免疫的PRN抗體,兩組的抗體滴度無(wú)顯著性差異(p0.05)。此結(jié)果表明重組PRN具有較好的免疫原性和抗原性。百日咳的攻擊實(shí)驗(yàn)表明,重組PRN免疫小鼠后可以產(chǎn)生一定的保護(hù)力,抵抗百日咳攻擊菌18323的感染。在三個(gè)免疫劑量中,中劑量的保護(hù)率最高。將重組PRN和天然PRN分別與百日咳疫苗原液、FIM2和FIM3組合模擬無(wú)細(xì)胞百日咳組分疫苗,進(jìn)行百日咳攻擊實(shí)驗(yàn)。結(jié)果表明,重組PRN+原液+FIM2+FIM3組高劑量的存活率和天然PRN+原液+FIM2+FIM3組高劑量的相同,均為最高值93.75%,而中劑量和低劑量的存活率高于天然PRN+原液+FIM2+FIM3組的。 細(xì)胞因子的檢測(cè)結(jié)果表明,重組PRN組分泌較高水平的Th2型細(xì)胞因子IL-4和IL-12,而Th1型細(xì)胞因子IL-2,IL-10和IFN-γ的分泌量較低,說(shuō)明重組PRN可以激發(fā)體液免疫和細(xì)胞免疫,但以體液免疫應(yīng)答為主。天然PRN組的應(yīng)答類(lèi)型與重組PRN組相同。模擬疫苗重組PRN+原液+FIM2+FIM3組和天然PRN+原液+FIM2+FM3組也以體液免疫為主要的應(yīng)答方式。 通過(guò)對(duì)IgGl和IgG2a的測(cè)定表明,重組PRN組、天然PRN組、重組PRN+原液+FIM2+FIM3組和天然PRN+原液+FIM2+FIM3組既產(chǎn)生IgGl又產(chǎn)生IgG2a,且IgGl高于IgG2a,說(shuō)明以上抗原可以激發(fā)機(jī)體產(chǎn)生一定程度的體液免疫和細(xì)胞免疫,但體液免疫為主要的免疫應(yīng)答類(lèi)型。 本文的研究標(biāo)明,重組PRN完全有可能代替天然PRN應(yīng)用于百日咳組分疫苗。此研究為研發(fā)新型的多組分百日咳疫苗提供了有效的實(shí)踐依據(jù)。
[Abstract]:PRN (pertussis adhesives) is an important component of acellular pertussis vaccine. The purification process of natural PRN is complex and the preparation cost is high. It is a trend to develop new pertussis vaccine with recombinant PRN instead of natural PRN. In this paper, the immunogenicity, antigenicity and protective power of recombinant PRN were evaluated, and the types of immune response induced by recombinant PRN were studied. The antibody titers of recombinant PRN and natural PRN were not significantly different from those of PRN group and natural PRN group. The results showed that the recombinant PRN had good immunogenicity and antigenicity. The results of pertussis attack test showed that there was no significant difference in the titer of PRN antibody between the two groups, and the results showed that the recombinant PRN had good immunogenicity and antigenicity. After immunizing mice with recombinant PRN, it can produce a certain protective effect against the infection of 18323 pertussis. The recombinant PRN and natural PRN were combined with pertussis vaccine extract (FIM2) and FIM3 to simulate the acellular pertussis component vaccine. The results showed that the recombinant PRN and natural PRN were used to simulate the acellular pertussis component vaccine. The high dose survival rate of FIM2 FIM3 group was the same as that of FIM2 FIM3 group, which was the highest value of 93.75%, but the survival rate of medium and low dose of FIM2 FIM3 group was higher than that of FIM2 FIM3 group. The results of cytokine detection showed that the recombinant PRN group secreted higher levels of Th2 type cytokines IL-4 and IL-12, while Th1 type cytokines IL-2, IL-10 and IFN- 緯 secreted less, suggesting that recombinant PRN could stimulate humoral and cellular immunity. But humoral immune response was dominant. The type of response in natural PRN group was the same as that in recombinant PRN group. Humoral immunity was also the main response mode in FIM2 FIM3 group and FIM2 FM3 group. The determination of IgGl and IgG2a showed that the recombinant PRN group, the natural PRN group, The recombinant PRN FIM2 FIM3 group and the natural PRN FIM2 FIM3 group produced both IgGl and IgG2a, and the IgGl was higher than IgG2a, indicating that the above antigens could stimulate humoral and cellular immunity to some extent, but humoral immunity was the main type of immune response. The research in this paper indicates that recombinant PRN can replace natural PRN in the application of pertussis component vaccine. This study provides an effective practical basis for the development of a new multicomponent pertussis vaccine.
【學(xué)位授予單位】：昆明理工大學(xué)
【學(xué)位級(jí)別】：碩士
【學(xué)位授予年份】：2009
【分類(lèi)號(hào)】：R392

【參考文獻(xiàn)】

相關(guān)期刊論文 前7條

1 肖詹蓉 ,劉�？�;百日咳疫苗及其免疫接種[J];國(guó)外醫(yī)學(xué)(預(yù)防、診斷、治療用生物制品分冊(cè));2004年05期

2 楊曉明,何長(zhǎng)民;百日咳桿菌粘著素的分子克隆、表達(dá)、純化及生物學(xué)特性研究[J];微生物學(xué)免疫學(xué)進(jìn)展;1997年04期

3 張興錄,楊志偉,周軍,于競(jìng)進(jìn),王克安;我國(guó)近年百日咳流行病學(xué)特點(diǎn)分析[J];中國(guó)計(jì)劃免疫;2000年02期

4 劉�？�,迮文遠(yuǎn);百日咳的免疫及預(yù)防[J];中國(guó)計(jì)劃免疫;2000年02期

5 符劍,王躍芳,徐寶祥,胡昱,郁子揚(yáng);浙江省1954~2004年百日咳流行病學(xué)分析[J];中國(guó)計(jì)劃免疫;2005年04期

6 于燕,刁琳琪,申振元;河南省1963~2002年百日咳流行動(dòng)態(tài)及控制策略分析[J];中國(guó)計(jì)劃免疫;2005年05期

7 孟成艷;張文宏;;全球百日咳疾病負(fù)擔(dān)的現(xiàn)狀與展望[J];中國(guó)計(jì)劃免疫;2006年04期

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