不同佐劑對(duì)含S+PreS1融合抗原的乙型肝炎病毒顆粒疫苗免疫原性的影響

陳紅，鄧瑤，譚文杰，王文，殷霄，管潔，王文玲，阮力

中國(guó)疾病預(yù)防控制中心病毒病預(yù)防控制所 病毒病應(yīng)急技術(shù)中心，北京 100052

不同佐劑對(duì)含S+PreS1融合抗原的乙型肝炎病毒顆粒疫苗免疫原性的影響

陳紅*，鄧瑤*，譚文杰，王文，殷霄，管潔，王文玲，阮力

中國(guó)疾病預(yù)防控制中心病毒病預(yù)防控制所 病毒病應(yīng)急技術(shù)中心，北京 100052

本研究在前期工作基礎(chǔ)上，用CHO細(xì)胞表達(dá)的含PreS1+S融合抗原的新型基因工程HBV顆粒疫苗(HBSS1)與Al(OH)3、CpG及CpG+Al(OH)3等佐劑配伍，在Balb/C小鼠模型上研究不同佐劑對(duì)HBV顆粒疫苗肌肉注射后免疫應(yīng)答的影響，主要包括抗體滴度、抗體亞型分類及特異性細(xì)胞免疫(γ-IFN ELISpot檢測(cè))。結(jié)果表明：CpG佐劑結(jié)合HBSS1顆粒疫苗可快速誘導(dǎo)(單針免疫)高水平的抗PreS1及S抗體，IgG2a/IgG1比率＞1，同時(shí)可誘導(dǎo)較高抗原特異的細(xì)胞免疫應(yīng)答；Al(OH)3+CpG雙佐劑組一次免疫后可誘導(dǎo)產(chǎn)生最高的抗S抗體滴度(1:105)，其產(chǎn)生的抗體亞類包括IgG1、IgG2a與IgG2b；在S抗原N端(13～49 aa)存在優(yōu)勢(shì)CTL表位。結(jié)論：CpG佐劑結(jié)合HBSS1顆粒疫苗應(yīng)是發(fā)展新型治療性乙肝疫苗的較佳選項(xiàng)。

乙型肝炎病毒，病毒顆粒，疫苗，CpG佐劑

Abstract:We previous reported the development of novel hepatitis B virus(HBV)vaccine containing the surface antigen(S)plus PreS1 fusion derived from Chinese hamster ovary(CHO)cells system.In this study, we analyzed the impact of different adjuvants on immunogenicity of the HBV particle vaccine in Balb/C mice, including alum alone, CpG oligodeoxynucleotides(CpG-ODN)alone and CpG-ODN in combination with alum adjuvant.We first detected the antigen specific humoral response in mice, including total IgG antibody and IgG subtyping.Then, we characterized the specific cell-mediated immune(CMI)response by detection of γ-interferon secreting splenocytes after stimulating with S or PreS1 peptide pool.Our results showed that: CpG-ODN adjuvanted vaccine could rapidly induce higher level of anti-PreS1 and anti-S antibodies, and a higher ratio of IgG2a/IgG1 antibody than that of alum adjuvanted vaccine.At the same time, CpG-ODN adjuvanted vaccine induced robust antigen-specific cellular immune responses in mice, which was superior to that of alum adjuvanted vaccine and CpG-ODN in combination with alum adjuvantedvaccine; however, the vaccine candidate with CpG-ODN in combination with alum adjuvant induced highest anti-S antibody and mixed IgG subclasses in mice after twice immunization.There exists dominant HBV CMI epitopes in the N-terminal of S antigen.These results provided important evidence that CpG-ODN adjuvanted HBSS1 particles vaccine may serve as a novel candidate in the development of new preventive and therapeutic agents against hepatitis B infection.

Keywords:hepatitis B virus(HBV), viral particles, vaccine, CpG-ODN

乙型肝炎病毒(HBV)不僅可引起急慢性肝炎，而且與肝硬化、肝癌的發(fā)生密切相關(guān)，盡管乙肝疫苗的使用已逾20年，目前HBV仍是傳播最為廣泛的病原之一[1]。據(jù) WHO 分析，全世界每年仍有 50～120萬(wàn)人死于HBV感染[1]。HBV疫苗在中國(guó)已納入計(jì)劃免疫，使用最為廣泛的為酵母或CHO細(xì)胞表達(dá)含S全長(zhǎng)的顆粒亞單位疫苗[2]。HBV疫苗長(zhǎng)期使用的效果表明，該疫苗加Al(OH)3佐劑后三針免疫可有效保護(hù)人群HBV感染與傳播，但仍有5%～10%的人群不能產(chǎn)生保護(hù)性anti-HBS抗體，且近年來(lái)，S變異流行株呈增長(zhǎng)趨勢(shì)[2]。……