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CpG對(duì)乙肝核酸疫苗及基因重組(CHO細(xì)胞)疫苗免疫效果的影響

發(fā)布時(shí)間:2018-02-24 02:32

  本文關(guān)鍵詞: CpG佐劑 乙肝核酸疫苗(DNA疫苗) 基因重組(CHO)乙肝疫苗 出處:《吉林大學(xué)》2007年碩士論文 論文類型:學(xué)位論文


【摘要】: 接種乙肝疫苗是預(yù)防和控制乙肝病毒傳播的重要措施。血源乙肝疫苗由于潛在安全性問題,現(xiàn)在已由基因工程疫苗所替代,但是,基因工程乙肝疫苗應(yīng)用過程中也存在一些問題,其中酵母苗成本低,母嬰阻斷效果好,但體液免疫水平較低,所以GMT水平低,造成許多接種者產(chǎn)生低滴度抗體(〈10IU/L〉或不產(chǎn)生抗體,平均發(fā)生率在20%左右;重組CHO細(xì)胞疫苗糖基化水平較好,但細(xì)胞免疫水平較低,生產(chǎn)成本高,同樣在全程免疫人群中,約有5%-10%的人不產(chǎn)生保護(hù)性抗體或抗體水平很低,老年人群比例尤高。現(xiàn)有疫苗也不能打破乙肝患者或乙肝病毒攜帶者機(jī)體的免疫耐受使之清除病毒,另外,對(duì)有可能逃避現(xiàn)有疫苗保護(hù)作用的HBsAg突變株存在爭(zhēng)議。 本文在構(gòu)建出乙肝核酸疫苗的基礎(chǔ)上,考察了CpG對(duì)乙肝核酸疫苗及基因重組乙肝疫苗免疫效果的影響。CpG- ODN作為佐劑可促進(jìn)HBV基因疫苗誘導(dǎo)Balb/c小鼠產(chǎn)生的細(xì)胞免疫應(yīng)答,其佐劑效應(yīng)主要由于CpG基序的存在。乙肝疫苗用量減少三倍時(shí),誘導(dǎo)抗體產(chǎn)生水平和小鼠陽轉(zhuǎn)數(shù)量皆無明顯改變,說明CpG對(duì)誘導(dǎo)體液免疫具有較好的增強(qiáng)作用。這可能是由于其增強(qiáng)了機(jī)體對(duì)乙肝基因重組疫苗的反應(yīng)敏感性,從而降低疫苗的ED50,減少了疫苗的用量CpG與基因重組(CHO)乙肝疫苗聯(lián)合免疫小鼠的實(shí)驗(yàn)中,無論從小鼠的陽轉(zhuǎn)率還是小鼠的陽轉(zhuǎn)時(shí)間均高于或早于CpG與乙肝核酸疫苗聯(lián)合免疫的實(shí)驗(yàn)。應(yīng)用CpG-寡聚脫氧核苷酸(CpG-ODN)與不同劑量HBV疫苗配伍分別免疫小鼠后抗-HBs陽轉(zhuǎn)率及抗-HBs水平分別較單用核酸疫苗的對(duì)照組高3倍和2倍。而CpG和重組HBV疫苗配伍免疫后抗-HBs陽轉(zhuǎn)率和陽轉(zhuǎn)時(shí)間均高于和早于CpG與核酸疫苗組。加入CpG-ODN后,即使重組HBV疫苗用量減少3倍,其誘導(dǎo)抗-HBs陽轉(zhuǎn)率和水平亦無改變。這樣有望在人類應(yīng)用上更有效、經(jīng)濟(jì)。
[Abstract]:Vaccination of hepatitis B vaccine is an important measure to prevent and control the spread of hepatitis B. due to potential safety problems, blood source hepatitis B vaccine has now been replaced by genetic engineering vaccine, but, There are also some problems in the application of genetically engineered hepatitis B vaccine, including low cost of yeast vaccine, good blocking effect of mother and child, but low level of humoral immunity, so the level of GMT is low, resulting in low titer antibody (< 10 IUP / L > or no antibody) produced by many inoculators. The average incidence was about 20%, the glycosylation level of recombinant CHO cell vaccine was good, but the level of cellular immunity was lower and the production cost was high. In the same population, about 5- 10% of the whole immune population did not produce protective antibody or antibody level was very low. The existing vaccine can not break the immune tolerance of hepatitis B patients or hepatitis B carriers to clear the virus. In addition, there is controversy about the HBsAg mutants that may escape the protective effect of the existing vaccine. Based on the construction of hepatitis B nucleic acid vaccine, the effect of CpG on the immune effect of hepatitis B nucleic acid vaccine and recombinant hepatitis B vaccine. CpG- ODN as adjuvant can promote the cellular immune response of Balb/c mice induced by HBV gene vaccine. The adjuvant effect was mainly due to the existence of CpG motif. When the dosage of hepatitis B vaccine decreased by three times, the level of antibody production and the number of positive transformation in mice did not change obviously. It is suggested that CpG can enhance humoral immunity, which may be due to its enhanced sensitivity to recombinant hepatitis B vaccine. Thus reducing the ED50 of the vaccine and reducing the dosage of the vaccine CpG and recombinant hepatitis B vaccine co-immunized mice, Both the positive conversion rate of mice and the positive transition time of mice were higher or earlier than that of CpG combined with hepatitis B nucleic acid vaccine. Anti-HBs were immunized with CpG- oligodeoxynucleotides (CpG-ODN) and different doses of HBV vaccine respectively. The positive conversion rate and anti-HBs level of CpG and recombinant HBV vaccine were 3 and 2 times higher than those of the control group, respectively, and the positive conversion rate and time of CpG and recombinant HBV vaccine were higher than and earlier than those of CpG and nucleic acid vaccine group. After the addition of CpG-ODN, the positive conversion rate and time of anti-HBs positive transformation were higher than those of CpG and nucleic acid vaccine group. Even if the dosage of recombinant HBV vaccine is reduced threefold, the positive conversion rate and level of anti-HBs induced by recombinant HBV vaccine will not change, which is expected to be more effective and economical in human application.
【學(xué)位授予單位】:吉林大學(xué)
【學(xué)位級(jí)別】:碩士
【學(xué)位授予年份】:2007
【分類號(hào)】:R392

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