本文關(guān)鍵詞：EV71滅活疫苗交叉保護(hù)性及其與甲肝聯(lián)合疫苗免疫效果的初步評(píng)價(jià)　出處：《北京協(xié)和醫(yī)學(xué)院》2012年碩士論文　論文類(lèi)型：學(xué)位論文

  更多相關(guān)文章： 腸道病毒 腸道病毒71型 交叉反應(yīng) 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(wú)菌性腦膜炎、急性馳緩性麻痹、腦干腦炎和其他一些嚴(yán)重的神經(jīng)系統(tǒng)疾病,包括肺水腫和心肺功能障礙。近年來(lái),我國(guó)EV71感染出現(xiàn)流行趨勢(shì),2008年國(guó)家已將手足口病納入丙類(lèi)傳染病管理,預(yù)防EV71感染、降低發(fā)病率成為重要的公共衛(wèi)生問(wèn)題。 疫苗接種是預(yù)防控制最有效的方法,中國(guó)醫(yī)學(xué)科學(xué)院醫(yī)學(xué)生物學(xué)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(hù)作用關(guān)系到疫苗的預(yù)防效果。為充分了解我所疫苗對(duì)于其他不同EV71毒株的交叉保護(hù)性,本研究應(yīng)用FY-23K-B株免疫血清對(duì)收集的K9、S1、R20-9J-3、S19、合49、BrCr等20個(gè)EV71不同毒株,同時(shí)對(duì)同屬的脊髓灰質(zhì)炎病毒(polio Ⅰ/Ⅱ/Ⅲ)、柯薩奇A組16型(CoxA16)、�？刹《�2/6型(ECHO2/6)等8株非EV71腸道病毒株進(jìn)行中和試驗(yàn)。結(jié)果表明FY-23K-B株制備的疫苗對(duì)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(yīng),而2U血清中和效力相對(duì)較低,只能中和約35%的毒株；沒(méi)有發(fā)現(xiàn)該株血清對(duì)其它8個(gè)非EV71的腸道病毒株的交叉保護(hù)作用。 開(kāi)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(jìn)行了研究。我們將EV71(EU/ml)/HAV (EU/ml)進(jìn)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(gè)聯(lián)苗組,免疫ICR小鼠,并與相同劑量的單價(jià)疫苗組進(jìn)行對(duì)比。初免后28天和加強(qiáng)后7天檢測(cè)機(jī)體的特異性體液免疫和細(xì)胞免疫應(yīng)答水平,結(jié)果發(fā)現(xiàn)初免后28天各組小鼠100%產(chǎn)生了保護(hù)性HAV抗體,160/640(EV71/HAV)組最高(4676.6mIU/ml),最低組為HAV80EU/ml單價(jià)組(131.5mIU/ml),加強(qiáng)后抗體平均增長(zhǎng)12.2倍,其中160/640(EV71/HAV)組高達(dá)28101.8mIU/ml,但該組與320/640、640/640組間差異無(wú)統(tǒng)計(jì)學(xué)意義(P0.05)。初免后血清中EV71抗體陽(yáng)轉(zhuǎn)率即達(dá)到100%,各組平均GMT值為14.5,并表現(xiàn)出劑量效應(yīng),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(wú)統(tǒng)計(jì)學(xué)意義(P0.05)。流式細(xì)胞術(shù)檢測(cè)發(fā)現(xiàn)初免后各組小鼠脾臟中CD8+細(xì)胞比例高于CD4+細(xì)胞(P0.001),加強(qiáng)后則相反(P0.001),而HAV全病毒和EV71多肽刺激脾細(xì)胞產(chǎn)生的陽(yáng)性反應(yīng)弱,加強(qiáng)后脾細(xì)胞分泌的IL-4高于IFN-γ(P0.05)。說(shuō)明一定劑量的HAV和EV71聯(lián)合免疫可誘導(dǎo)機(jī)體產(chǎn)生良好的體液免疫應(yīng)答,但細(xì)胞免疫應(yīng)答較弱,就體液免疫應(yīng)答而言,我們認(rèn)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(xué)位授予單位】：北京協(xié)和醫(yī)學(xué)院
【學(xué)位級(jí)別】：碩士
【學(xué)位授予年份】：2012
【分類(lèi)號(hào)】：R392

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