【摘要】： 隨著全球消滅脊髓灰質(zhì)炎(脊灰)目標(biāo)的臨近,疫苗衍生脊灰病毒(Vaccine-derived Poliovirus, VDPV)的流行、疫苗相關(guān)麻痹型病例(Vaccine Associated Paralytic Poliomyelitis, VAPP)的發(fā)生等對(duì)無脊灰狀態(tài)成果的鞏固威脅很大。在消滅脊灰的最后階段,應(yīng)盡快消滅脊灰野毒株,并使用脊髓灰質(zhì)炎滅活疫苗(Inactivated poliovirus vaccine, IPV)替代目前廣泛使用的脊髓灰質(zhì)炎減毒活疫苗(Oral poliovirus vaccine, OPV)來維持無脊灰狀態(tài),消除自然界的活病毒�？紤]到疫苗生產(chǎn)的生物安全性等因素,WHO鼓勵(lì)研制Sabin株脊髓灰質(zhì)炎滅活疫苗(Sabin IPV)。 本論文旨在針對(duì)Sabin II型病毒,尋找一種與傳統(tǒng)滅活劑-甲醛相比較,對(duì)病毒的抗原破壞更小,且更安全的病毒滅活劑；并進(jìn)一步探索Sabin IPV中添加佐劑對(duì)免疫效果產(chǎn)生的影響,探討Sabin IPV最適劑量和佐劑應(yīng)用的意義。 本研究第一部分進(jìn)行了二乙烯亞胺(BEI)對(duì)脊髓灰質(zhì)炎病毒Ⅱ型Sabin株的滅活效果觀察。通過在不同溫度下采用4種濃度的BEI及甲醛對(duì)脊髓灰質(zhì)炎病毒Ⅱ型Sabin株進(jìn)行滅活,測(cè)定活病毒含量和D抗原含量,并進(jìn)行病毒滅活驗(yàn)證試驗(yàn)。結(jié)果在37℃條件下,0.002 mol/L濃度的BEI能在48h內(nèi)完全滅活病毒,滅活后的病毒液D抗原含量回收率為90.6%,顯著高于甲醛滅活的回收率(60.6%)。經(jīng)驗(yàn)證滅活徹底,無活病毒存在。BEI對(duì)脊髓灰質(zhì)炎病毒Ⅱ型Sabin株的D抗原破壞小,能較好的保持疫苗的有效成分。 本研究的第二部分四組不同劑量的Sabin IPV疫苗與佐劑Al(OH)3配伍后免疫Wistar大鼠,通過檢測(cè)各組大鼠免疫后血清中和抗體水平、特異性IFN-γ的產(chǎn)生情況以及免疫后各組大鼠CD3/CD4/CD8分子表達(dá)的差異,以確定使用Al(OH)3佐劑時(shí)Sabin IPV的最適劑量。通過微量中和實(shí)驗(yàn)檢測(cè)Sabin株脊髓灰質(zhì)炎特異性中和抗體發(fā)現(xiàn),疫苗不同劑量產(chǎn)生的中和抗體GMT水平基本是抗原含量高的組高于抗原含量低的組,添加Al(OH)3佐劑組的中和抗體水平也是高于未添加佐劑組的中和抗體水平。ELISPOT測(cè)定針對(duì)Sabin株脊髓灰質(zhì)炎病毒的特異性IFN-γ的結(jié)果顯示,添加佐劑實(shí)驗(yàn)組產(chǎn)生的斑點(diǎn)要多于未添加佐劑組,2-①實(shí)驗(yàn)組三個(gè)型別所產(chǎn)生的斑點(diǎn)數(shù)都最多。流式細(xì)胞儀檢測(cè)各組大鼠CD3/CD4/CD8分子表達(dá)差異的結(jié)果顯示,1-①實(shí)驗(yàn)組CD3+CD4+/CD3+CD8+比值最高。綜上所述,添加佐劑對(duì)三個(gè)型別的SabinIPV的免疫反應(yīng)有增強(qiáng)的效果；綜合實(shí)驗(yàn)結(jié)果可認(rèn)為Ⅰ型采用30 DU/dose,Ⅱ型采用16 DU/dose,Ⅲ型22.5 DU/dose劑量時(shí)較為適宜。
[Abstract]:As the global goal of polio eradication approaches, the prevalence of vaccine-derived poliovirus (Vaccine-derived Poliovirus, VDPV) and the occurrence of vaccine-associated paralytic cases (Vaccine Associated Paralytic Poliomyelitis, VAPP) pose a great threat to the consolidation of poliomyelitis (poliomyelitis) status. In the final stage of polio eradication, the polio field strain should be eliminated as soon as possible, and poliomyelitis inactivated vaccine (Inactivated poliovirus vaccine, IPV) should be used to replace the widely used live attenuated poliomyelitis vaccine (Oral poliovirus vaccine, OPV) to maintain poliomyelitis free. Eliminate live viruses from nature. Who encourages the development of Sabin strain inactivated poliomyelitis vaccine (Sabin IPV). Considering the biological safety of vaccine production. The purpose of this paper is to search for a virus inactivated agent with less antigenic damage and more safety to Sabin II virus than traditional inactivated agent-formaldehyde, and to explore the effect of adjuvant added in Sabin IPV on the immune effect. To explore the significance of optimal dosage and adjuvant application of Sabin IPV. In the first part of this study, the inactivation effect of diethylenimide (BEI) on poliovirus type 鈪，

本文編號(hào)：2238579