本文選題：輪狀病毒疫苗 + 免疫原性�。� 參考：《北京協(xié)和醫(yī)學(xué)院》2016年博士論文

【摘要】：輪狀病毒(Rotavirus,RV)是導(dǎo)致世界范圍嬰幼兒重癥腹瀉的主要病原體之一,輪狀病毒感染被稱為“民主病”,無(wú)論發(fā)達(dá)還是發(fā)展中國(guó)家兒童都會(huì)感染,給世界各國(guó)帶來(lái)了嚴(yán)重的社會(huì)和經(jīng)濟(jì)負(fù)擔(dān)。但在發(fā)展中國(guó)家引起的5歲以下兒童死亡率更高。疫苗接種是唯一預(yù)防輪狀病毒腹瀉,降低死亡率的有效途徑。近十年來(lái),葛蘭素史克公司生產(chǎn)的人類輪狀病毒減毒活疫苗(Rotarix)和默沙東公司生產(chǎn)的口服輪狀病毒五價(jià)活疫苗(RotaTeq)在世界范圍廣泛使用。疫苗臨床試驗(yàn)和上市后調(diào)查數(shù)據(jù)顯示輪狀病毒減毒活疫苗在發(fā)達(dá)國(guó)家具有高效的保護(hù)作用,然而在最需要它們的發(fā)展中國(guó)家,疫苗的有效性和免疫原性卻不理想。對(duì)于這種現(xiàn)象,亟待找出原因以提高輪狀病毒疫苗在發(fā)展中國(guó)家的有效性。學(xué)者們針對(duì)此現(xiàn)象曾經(jīng)進(jìn)行過(guò)研究,從接種者的營(yíng)養(yǎng)狀況、腸道菌群、母乳喂養(yǎng)行為以及病毒株流行的不同入手,解釋免疫原性差異的原因。但這些闡述綜合起來(lái)分析并不能夠給出全面系統(tǒng)合理的解釋,也有待于從不同側(cè)面進(jìn)行更深入的研究。免疫系統(tǒng)具有復(fù)雜的網(wǎng)絡(luò)結(jié)構(gòu),導(dǎo)致輪狀病毒疫苗在不同地區(qū)免疫原性和有效性不同的也可能不僅限于上述幾個(gè)因素。鑒于此,本研究從接種者機(jī)體內(nèi)免前母?jìng)骺贵w、人類白細(xì)胞抗原(Human Leukocyte Antigen, H LA)基因多態(tài)性和同時(shí)接種口服脊髓灰質(zhì)炎病毒減毒活疫苗(Oral Poliovirus Vaccine, OPV)的行為三個(gè)方面分析可能影響輪狀病毒減毒活疫苗的免疫反應(yīng)的因素。本研究中,6-13周齡健康嬰幼兒接種三劑Ⅲ價(jià)輪狀病毒基因重配減毒活疫苗,每劑間隔1個(gè)月,分別在首劑接種前、第3劑接種后1個(gè)月采集血樣,進(jìn)行G2、G3、G4型特異性輪狀病毒免疫球蛋白A (Rotavirus-Immunoglobulin A, RV-IgA)抗體測(cè)定。分別根據(jù)各型免后RV-IgA是否陽(yáng)轉(zhuǎn),將接種人群分為各型血清陽(yáng)轉(zhuǎn)組和血清未陽(yáng)轉(zhuǎn)組。同時(shí)檢測(cè)接種人群的免前血清G2、G3、G4型特異性輪狀病毒中和抗體水平。統(tǒng)計(jì)各型中和抗體陽(yáng)性率和抗體水平在兩組間是否有差異。結(jié)果發(fā)現(xiàn)：接種人群中免前3個(gè)型特異性的輪狀病毒中和抗體陽(yáng)性率和抗體幾何平均滴度分別為G4型最低(49.24%,12),G2型居中(82.95%,20),G3型最高(100%,91)。與此相對(duì)應(yīng)地,群體免后呈現(xiàn)出RV-IgA的血清抗體水平G3型(76U/mL)最低,G2型(86U/mL)居中,G4型(90U/mL)最高的分布特點(diǎn)。免前各型特異性中和抗體的陽(yáng)性率和抗體水平均為血清陽(yáng)轉(zhuǎn)組低于血清未陽(yáng)轉(zhuǎn)組,除G3型外,G2和G4型免前中和抗體的陽(yáng)性率和抗體水平在兩組之間均有統(tǒng)計(jì)學(xué)差異(P0.05)。提示母?jìng)骺贵w可能會(huì)抑制輪狀病毒減毒活疫苗的免疫反應(yīng)。根據(jù)接種者首次接種Ⅲ價(jià)輪狀病毒基因重配減毒活疫苗時(shí)的年齡,將其分為兩組,平均年齡分別為8周齡和12周齡,統(tǒng)計(jì)各型免前中和抗體和免后RV-IgA在兩組間的差異,分析推遲免疫接種程序是否能夠消除母?jìng)骺贵w的影響,增強(qiáng)疫苗免疫反應(yīng)。與8周齡人群相比,在12周齡接種人群中,除G2型免前中和抗體陽(yáng)性率略高(83.19%：82.76%),以及G3型陽(yáng)性率均為100%外,免前其它型中和抗體陽(yáng)性率和抗體水平均較低,G3型抗體水平差異有統(tǒng)計(jì)學(xué)意義(P=0.003)。對(duì)于免后RV-IgA,在12周齡組中,除G3型RV-IgA的幾何平均滴度略低(76：77)外,其它型免后RV-IgA陽(yáng)轉(zhuǎn)率和抗體水平均較8周齡組高,但差異無(wú)統(tǒng)計(jì)學(xué)意義。提示母?jìng)骺贵w隨著時(shí)間的推移逐漸衰減,但在我國(guó)將疫苗接種時(shí)間推遲至12周齡,實(shí)際上對(duì)消除母?jìng)骺贵w的影響有一定的幫助,但也需要更多年齡組的研究。在上述接種人群的基礎(chǔ)上,將免后任何型RV-IgA陽(yáng)轉(zhuǎn)的接種者歸為血清陽(yáng)轉(zhuǎn)組,所有型別RV-IgA均未陽(yáng)轉(zhuǎn)的歸為血清未陽(yáng)轉(zhuǎn)組,分別從兩組中隨機(jī)選取55人和41人通過(guò)測(cè)序法進(jìn)行HLA-A、-B、-C、-DRB1、-DQB1分型,計(jì)算HLA各位點(diǎn)等位基因型別、單體型和超型在兩組人群中的分布頻數(shù),統(tǒng)計(jì)兩組間等位基因、單體型和超型的分布是否有差異,分析HLA基因多態(tài)性與輪狀病毒減毒活疫苗免疫反應(yīng)的關(guān)系。結(jié)果發(fā)現(xiàn)HLA-B*40:01在血清未陽(yáng)轉(zhuǎn)組的分布頻率明顯高于血清陽(yáng)轉(zhuǎn)組,差異有統(tǒng)計(jì)學(xué)意義,與輪狀病毒減毒活疫苗接種后無(wú)免疫應(yīng)答相關(guān)。為了評(píng)價(jià)同時(shí)接種OPV的行為是否會(huì)影響輪狀病毒減毒活疫苗的免疫反應(yīng)和血清陽(yáng)轉(zhuǎn)率,本研究以葛蘭素史克公司生產(chǎn)的Rotarix作為研究對(duì)象,將受試人群隨機(jī)分為兩組。一組按照常規(guī)兩劑接種程序完成兩劑口服Rotarix的接種,并根據(jù)國(guó)家規(guī)劃接種年齡接種計(jì)劃免疫的OPV,但與Rotarix不在同一天接種。另一組在按照常規(guī)接種程序完成兩劑口服Rotarix的接種基礎(chǔ)上,同時(shí)接種OPV。分別在首劑接種前、第2劑Rotarix接種后1個(gè)月采集血樣,進(jìn)行RV-IgA抗體測(cè)定。通過(guò)統(tǒng)計(jì)分析兩組的RV-IgA血清陽(yáng)轉(zhuǎn)率和水平分布是否有統(tǒng)計(jì)學(xué)差異。結(jié)果顯示間隔接種和同時(shí)接種組的血清RV-IgA陽(yáng)轉(zhuǎn)率分別為73.84%和63.95%,差異有統(tǒng)計(jì)學(xué)意義(P=0.033);兩組的免后RV-IgA抗體幾何平均數(shù)分別為97和90,差異無(wú)統(tǒng)計(jì)學(xué)意義(P0.05)。說(shuō)明同時(shí)接種口服脊髓灰質(zhì)炎減毒活疫苗可能會(huì)影響輪狀病毒減毒活疫苗的血清陽(yáng)轉(zhuǎn)率,但尚未發(fā)現(xiàn)其對(duì)群體的抗體水平的影響。與同時(shí)接種組相比,間隔接種組免后1年血清RV-IgA陽(yáng)轉(zhuǎn)率和幾何平均滴度均較高,差異有統(tǒng)計(jì)學(xué)意義(P0.05)。同時(shí)接種組在免后1年中的抗體陽(yáng)轉(zhuǎn)率和GMT下降更為明顯,提示同時(shí)接種對(duì)RV-IgA抗體水平的維持也有影響。綜上所述,本研究結(jié)果表明宿主母?jìng)骺贵w、宿主基因多態(tài)性和OPV都可能是影響輪狀病毒減毒活疫苗免疫原性的因素,對(duì)輪狀病毒疫苗的研發(fā)、臨床試驗(yàn)的設(shè)計(jì)和免疫程序的制定有一定的參考價(jià)值。
[Abstract]:Rotavirus (RV) is one of the major pathogens causing severe diarrhoea in infants worldwide. Rotavirus infection is called "democratic disease". Both developed and developing children will be infected and bring serious social and economic burden to all countries in the world. But the mortality rate of children under 5 years old in developing countries is caused by developing countries. Higher. Vaccination is the only effective way to prevent rotavirus diarrhea and reduce mortality. In the past ten years, the human rotavirus vaccine (Rotarix) produced by the GlaxoSmithKline Co and the oral rotavirus five valence live vaccine (RotaTeq) produced by Mercedes are widely used worldwide. The results show that the rotavirus attenuated live vaccine has an effective protective effect in developed countries. However, the effectiveness and immunogenicity of the vaccine are not ideal in the developing countries which need them most. For this phenomenon, it is urgent to find the reasons to improve the effectiveness of the rotavirus vaccine in the developing country. A study was carried out to explain the reasons for the difference in immunogenicity from the nutritional status of the inoculant, the intestinal flora, the breastfeeding behavior and the prevalence of the virus strains. However, the comprehensive analysis was not able to give a comprehensive and systematic explanation, and a more in-depth study of the immune system was needed. The heterogeneous network structure leads to the difference in the immunogenicity and effectiveness of the rotavirus vaccine in different regions, which may not only be limited to the above factors. In view of this, the present study is based on the polymorphism of the prefree maternal antibody, Human Leukocyte Antigen, H LA gene and the simultaneous inoculation of oral poliomyelitis virus in the inoculant. Three aspects of the behavior of Oral Poliovirus Vaccine (OPV) were analyzed in this study. In this study, 6-13 weeks old healthy infants were inoculated with three doses of rotavirus gene redistribution attenuated live vaccine, each interval of 1 months, before the first inoculation and 1 after third doses of inoculation, respectively. Blood samples were collected for G2, G3, and G4 specific rotavirus immunoglobulin A (Rotavirus-Immunoglobulin A, RV-IgA) antibody determination in the month. The inoculated crowd was divided into different seropositive groups and serum unpositive rotation groups according to the positive changes of each type of RV-IgA, and the pre free serum G2, G3, G4 type rotavirus was detected in the inoculated population. The positive rate of neutralization antibody and antibody level were different between the two groups. The results showed that the positive rate of neutralization antibody and the geometric mean titer of the first 3 types of rotavirus were the lowest (49.24%, 12), G2 type (82.95%, 20), and the highest G3 type (100%, 91) in the inoculated population. The serum antibody level of RV-IgA (76U/mL) was the lowest, G2 type (86U/mL) was in the middle and the highest distribution of G4 type (90U/mL). The positive rate and antibody level of the specific neutralizing antibody were lower than that of the serum non positive group, and the positive rate and antibody of the G2 and G4 type pre neutralizing antibody and antibody, except G3 type, were all the lowest. The level was statistically different between the two groups (P0.05). It suggested that the maternal antibody may inhibit the immune response of the rotavirus attenuated live vaccine. According to the age of the inoculant, the age of the first inoculation of the rotavirus gene and the live attenuated vaccine, it was divided into two groups. The average age was divided into 8 weeks and 12 weeks of age. The difference between the two groups of the two groups and the postponement of the immunization program could eliminate the influence of the maternal antibody and enhance the immunization response. Compared with the 8 week old population, the positive rate of the G2 type preempting neutralization antibody was slightly higher (83.19%:82.76%), and the positive rate of the G3 type was all outside the other type in the 12 week old population. The positive rate of antibody and the level of antibody were low, and the difference of G3 type antibody level was statistically significant (P=0.003). For the post free RV-IgA, in the 12 week old group, except the geometric mean titer of G3 type RV-IgA was slightly lower (76:77), the other type of RV-IgA positive conversion rate and antibody level were higher than those of the 8 week old group, but the difference was not statistically significant. The time lapse gradually attenuated, but in our country the delay of vaccination to 12 weeks was actually helpful to eliminate the influence of maternal antibody, but it also needed more age groups. On the basis of the above inoculation population, any type of RV-IgA positive inoculant was classified as serological positive group, and all types of RV-IgA were not positive. HLA-A, -B, -C, -DRB1, -DQB1 typing were randomly selected from the two groups, and the distribution frequency of HLA, monotype and super type in the two groups was calculated, and the distribution of the two groups was statistically different between the two groups, and the analysis of the HLA base was analyzed. The relationship between polymorphism and the immune response of rotavirus attenuated live vaccine was found. The results showed that the distribution frequency of HLA-B*40:01 in the non positive seroconversion group was significantly higher than that in the serological positive group. The difference was statistically significant, and it was not related to the immune response after the rotavirus attenuated live vaccine. In order to evaluate the effect of the behavior of OPV at the same time, it could affect the rotaform disease. The immune response and serum positive rate of the live attenuated vaccine were studied in this study with the Rotarix produced by GlaxoSmithKline Co. The subjects were randomly divided into two groups. One group was inoculated with two doses of Rotarix in accordance with the routine two doses of inoculation, and OPV was immunized by the national plan for inoculation of age inoculation, but not with Rotarix. The other group was inoculated on the same day. The other group was inoculated on the basis of two doses of oral Rotarix according to the routine inoculation procedure. At the same time, the blood samples were collected by OPV. before the first inoculation and 1 months after the second doses of Rotarix. The serum positive rate and the level distribution of the two groups of RV-IgA were statistically analyzed. The RV-IgA positive rates of serum RV-IgA in the interval inoculation group and the simultaneous inoculation group were 73.84% and 63.95%, respectively, and the difference was statistically significant (P=0.033). The geometric mean of RV-IgA antibody in the two groups was 97 and 90, respectively, and the difference was not statistically significant (P0.05). The serum positive rate of the live vaccine was not found to affect the antibody level of the group. Compared with the same group, the serum RV-IgA positive rate and the geometric mean titer were higher in the interval 1 years after the vaccination group, and the difference was statistically significant (P0.05). At the same time, the positive rate of antibody and the decrease of GMT in the inoculated group were more obvious, suggesting that the antibody positive rate and the decrease of the antibody were more obvious. At the same time, inoculation also has an impact on the maintenance of RV-IgA antibody level. To sum up, the results of this study suggest that host maternal antibody, host gene polymorphism and OPV may be factors affecting the immunogenicity of the rotavirus attenuated live vaccine, and have a certain reference price for the development of rotavirus vaccine, the design of clinical trials and the formulation of immune procedures. Value.
【學(xué)位授予單位】：北京協(xié)和醫(yī)學(xué)院
【學(xué)位級(jí)別】：博士
【學(xué)位授予年份】：2016
【分類號(hào)】：R392

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6 夏樂(lè)樂(lè);何彪;楊凡力;徐琳;涂長(zhǎng)春;;蝙蝠輪狀病毒分離與鑒定[A];中國(guó)畜牧獸醫(yī)學(xué)會(huì)家畜傳染病學(xué)分會(huì)第八屆全國(guó)會(huì)員代表大會(huì)暨第十五次學(xué)術(shù)研討會(huì)論文集[C];2013年

7 時(shí)洪艷;馮力;孫東波;;輪狀病毒引起的仔豬腹瀉[A];中國(guó)畜牧獸醫(yī)學(xué)會(huì)畜牧獸醫(yī)生物技術(shù)學(xué)分會(huì)暨中國(guó)免疫學(xué)會(huì)獸醫(yī)免疫分會(huì)第六次研討會(huì)論文集[C];2005年

8 吳淑霞;李秀峰;席靈娟;;急性腹瀉患兒糞便中A群輪狀病毒的檢測(cè)結(jié)果分析[A];第一屆寧夏醫(yī)學(xué)會(huì)檢驗(yàn)學(xué)分會(huì)學(xué)術(shù)年會(huì)論文匯編[C];2005年

9 胡秀華;何苗;;定量測(cè)定輪狀病毒滴度的幾種方法的優(yōu)化及細(xì)胞系的選擇[A];中醫(yī)藥生物化學(xué)與分子生物學(xué)通訊[C];2008年

10 王斌;孫紅;朱軍生;;PAGE法與膠體金法檢測(cè)兒童腹瀉輪狀病毒研究[A];湖北省微生物學(xué)會(huì)第十屆理事會(huì)分析微生物專業(yè)委員會(huì)第四次學(xué)術(shù)會(huì)議論文匯編[C];2006年

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10 文文;嚴(yán)把嬰幼兒“入口”衛(wèi)生關(guān)[N];廣東科技報(bào);2008年

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