本文關鍵詞：EV71滅活疫苗交叉保護性及其與甲肝聯(lián)合疫苗免疫效果的初步評價　出處：《北京協(xié)和醫(yī)學院》2012年碩士論文　論文類型：學位論文

  更多相關文章： 腸道病毒 腸道病毒71型 交叉反應 甲型肝炎病毒 滅活疫苗 聯(lián)合免疫

【摘要】：腸道病毒71型(Enterovirus71, EV71)是手足口病(hand, foot and mouth disease, HFMD)的主要病原體,病毒感染多發(fā)于嬰幼兒,表現(xiàn)為手足口病、無菌性腦膜炎、急性馳緩性麻痹、腦干腦炎和其他一些嚴重的神經系統(tǒng)疾病,包括肺水腫和心肺功能障礙。近年來,我國EV71感染出現(xiàn)流行趨勢,2008年國家已將手足口病納入丙類傳染病管理,預防EV71感染、降低發(fā)病率成為重要的公共衛(wèi)生問題。 疫苗接種是預防控制最有效的方法,中國醫(yī)學科學院醫(yī)學生物學研究所篩選出EV71FY-23K-B株作為毒種研制成人用滅活疫苗。由于EV71存在不同的亞型,不同株、不同亞型間是否有交叉保護作用關系到疫苗的預防效果。為充分了解我所疫苗對于其他不同EV71毒株的交叉保護性,本研究應用FY-23K-B株免疫血清對收集的K9、S1、R20-9J-3、S19、合49、BrCr等20個EV71不同毒株,同時對同屬的脊髓灰質炎病毒(polio Ⅰ/Ⅱ/Ⅲ)、柯薩奇A組16型(CoxA16)、埃可病毒2/6型(ECHO2/6)等8株非EV71腸道病毒株進行中和試驗。結果表明FY-23K-B株制備的疫苗對于A型株和不同的C4亞型株表現(xiàn)出廣譜抗原性,其中4U(中和抗體)的特異性免疫血清已具有顯著的中和效應,而2U血清中和效力相對較低,只能中和約35%的毒株；沒有發(fā)現(xiàn)該株血清對其它8個非EV71的腸道病毒株的交叉保護作用。 開發(fā)聯(lián)合疫苗是疫苗發(fā)展的方向,為此我們就EV71與甲型肝炎病毒(Hepatitis A virus,HAV)滅活疫苗聯(lián)合免疫的可行性進行了研究。我們將EV71(EU/ml)/HAV (EU/ml)進行不同的組合成為640/320、640/160、640/80、160/1280、160/640、320/640、640/640共7個聯(lián)苗組,免疫ICR小鼠,并與相同劑量的單價疫苗組進行對比。初免后28天和加強后7天檢測機體的特異性體液免疫和細胞免疫應答水平,結果發(fā)現(xiàn)初免后28天各組小鼠100%產生了保護性HAV抗體,160/640(EV71/HAV)組最高(4676.6mIU/ml),最低組為HAV80EU/ml單價組(131.5mIU/ml),加強后抗體平均增長12.2倍,其中160/640(EV71/HAV)組高達28101.8mIU/ml,但該組與320/640、640/640組間差異無統(tǒng)計學意義(P0.05)。初免后血清中EV71抗體陽轉率即達到100%,各組平均GMT值為14.5,并表現(xiàn)出劑量效應,其中640/80(EV71/HAV)組抗體水平最高,但其與640/640、640/320、640/160組間差別無統(tǒng)計學意義(P0.05)。流式細胞術檢測發(fā)現(xiàn)初免后各組小鼠脾臟中CD8+細胞比例高于CD4+細胞(P0.001),加強后則相反(P0.001),而HAV全病毒和EV71多肽刺激脾細胞產生的陽性反應弱,加強后脾細胞分泌的IL-4高于IFN-γ(P0.05)。說明一定劑量的HAV和EV71聯(lián)合免疫可誘導機體產生良好的體液免疫應答,但細胞免疫應答較弱,就體液免疫應答而言,我們認為640/640(EV71/HAV)組的劑量配比是比較理想的；未發(fā)現(xiàn)兩種抗原間存在相互干擾或協(xié)同作用。
[Abstract]:Enterovirus 71 (Enterovirus71, EV71) is HFMD (hand foot, and mouth disease, HFMD) of the main pathogens, virus infection in infants, manifested as HFMD, aseptic meningitis, acute flaccid paralysis, brain stem encephalitis and other serious neurological diseases, including pulmonary edema and heart and lung dysfunction. In recent years, China's EV71 infection epidemic trend, 2008 has been included HFMD class C infectious disease management and prevention of EV71 infection, reduce the incidence become an important public health problem.
Vaccination is the most effective method of prevention and control, medical and Biological Research Academy of Medical Sciences Chinese selected EV71FY-23K-B strain of virus inactivated vaccine development for adult. Due to the existence of EV71 subtype, different strains, whether there is cross protection in relation to the vaccine prevention effect in different subtypes. In order to fully understand what I vaccine for cross protection of different EV71 strains, FY-23K-B strains used in this study to collect the serum K9, S1, R20-9J-3, S19, 49 BrCr, 20 EV71 strains, also belong to the polio virus (polio I / II / III), coxsackievirus group A type 16 (CoxA16), ECHO virus type 2/6 (ECHO2/6) and 8 strains of non EV71 strains of intestinal virus neutralization test. The results showed that FY-23K-B strain vaccine prepared for type A strains and different C4 subtype strains showed broad-spectrum antigenicity, 4U (neutralizing antibody) specific immune Serum has a significant neutralization effect, while the neutralizing potency of 2U is relatively low, which only neutralizes about 35% of the strains. No cross protection effect of the serum on other 8 non EV71 enterovirus strains was found.
The development of the combined vaccine is a vaccine development direction, therefore we EV71 and hepatitis A virus (Hepatitis A virus, HAV) inactivated vaccine combined with immune feasibility is studied. We will EV71 (EU/ml) /HAV (EU/ml) by different combinations of 640/320640/160640/ 80160/1280160/640320/640640/640 a total of 7 vaccine group, ICR mice were immunized at and with the same dose of vaccine group were compared. After immunization 28 days and 7 days after the body to strengthen detection of specific humoral and cellular immune response level, results showed that 28 days after immunization of mice produced 100% protective HAV antibody, 160/640 (EV71/HAV) was the highest (4676.6mIU/ml), HAV80EU/ml group was the lowest the unit group (131.5mIU/ml), anti average growth of 12.2 times, including 160/640 (EV71/HAV) group up to 28101.8mIU/ml, but there was no significant difference between the group and the 320/640640/640 group (P0.05) . after immunization of serum EV71 antibody positive rate was 100%, the average GMT value of 14.5, and showed a dose effect of 640/80 (EV71/HAV) group, the highest antibody level, but the difference between the 640/640640/320640/160 group had no statistical significance (P0.05). Flow cytometry found after immunization of CD8+ cells the proportion of mice spleen cells was higher than that of CD4+ (P0.001), strengthen the contrary (P0.001), and the positive reaction of HAV virus and EV71 polypeptide stimulated spleen cells produced weak spleen cells secrete IL-4 after strengthening is higher than that of IFN- gamma (P0.05). A dose of HAV and EV71 combined immunization can induce good humoral immune responses, but the cellular immune response is weak, humoral immune response, we believe that the 640/640 (EV71/HAV) dose ratio group is ideal; not found two antigens exist between mutual interference and synergistic effect.

【學位授予單位】：北京協(xié)和醫(yī)學院
【學位級別】：碩士
【學位授予年份】：2012
【分類號】：R392

【參考文獻】

相關期刊論文 前10條

1 李明;尹怡;李晉濤;;EV71疫苗的研究進展[J];生物醫(yī)學工程學雜志;2010年04期

2 任軍;李建民;陳薇;;聯(lián)合疫苗應用現(xiàn)狀及評價[J];中國生物工程雜志;2009年11期

3 劉建生,邵聰文,孟明耀,馬進,白巍,侯宗柳,陳統(tǒng)球;甲型肝炎滅活疫苗抗原劑量的優(yōu)化[J];中國生物制品學雜志;2005年05期

4 蘭蕓;胡凝珠;胡云章;瞿素;段金梅;李藍江;;3種培養(yǎng)方法培養(yǎng)脊髓灰質炎病毒過程中病毒繁殖特性比較[J];中國生物制品學雜志;2006年06期

5 王晶晶;趙紅玲;張瑩;劉龍丁;王麗春;廖蕓;董承紅;納銳雄;張雪梅;李琦涵;;腸道病毒71型滅活疫苗免疫恒河猴在攻毒實驗中的感染動力學[J];微生物與感染;2010年04期

6 李艷萍,李榮成,楊進業(yè),黃秋明,農藝,黃貴彪,龔健,潘慶琪,黃月葵,萬宗舉,劉崇柏;成人和兒童接種國產甲型肝炎滅活疫苗的安全性與免疫原性的初步觀察[J];中國計劃免疫;2003年04期

7 鄭徽;崔富強;;甲型肝炎減毒活疫苗與滅活疫苗免疫原性及其影響因素[J];中國疫苗和免疫;2009年04期

8 朵建英;王衛(wèi);馬春梅;郝翊;徐艷峰;許文波;董小平;王健偉;魏強;秦川;;EV71小鼠適應株制備及體內外感染特點[J];中國實驗動物學報;2010年05期

9 張春濤;劉春雨;吳瑜;趙晨燕;王佑春;;ELISPOT,MHC肽五聚體和ICCD三種檢測特異性細胞免疫應答方法的比較研究[J];中華實驗和臨床病毒學雜志;2006年04期

10 梁爭論;;腸道病毒71型疫苗候選株和疫苗標準的研究[J];中國病毒病雜志;2011年01期

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