INDUCTION OF PROTECTIVE CLASS-I MHC-RESTRICTED CTL IN MICE BY A RECOMBINANT INFLUENZA VACCINE IN ALUMINUM HYDROXIDE ADJUVANT
INDUCTION OF PROTECTIVE CLASS-I MHC-RESTRICTED CTL IN MICE BY A RECOMBINANT INFLUENZA VACCINE IN ALUMINUM HYDROXIDE ADJUVANT
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DOI：
10.1016/0264-410x(92)90369-u
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发表时间：
1992-01-01
期刊：
影响因子：
5.5
通讯作者：
HANNA, N
中科院分区：
文献类型：
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作者：
DILLON, SB;DEMUTH, SG;HANNA, N
通过可溶性蛋白质或肽诱导I类MHC限制的细胞毒性T淋巴细胞（CTL）应答需要复杂的佐剂或载体系统，其未被许可用于人疫苗。本报告中提供的数据表明，用氢氧化铝佐剂中的高度纯化的重组流感蛋白抗原接种疫苗（目前批准用于临床使用的唯一佐剂），可诱导I类限制性CTL，并保护免受H1N1和H2 N2病毒的致死性攻击。抗原（D蛋白，SK&F 106160）通过在大肠杆菌中表达H1N1流感病毒衍生的cDNA（菌株A/PR/8/34）而产生，并且由非结构蛋白1（NS 1）的前81个N-末端氨基酸（aa）通过9个核苷酸的非病毒接头序列融合至血凝素2亚基（HA 2）的157个C-末端aa组成。Kuwano等人1先前的工作表明，用部分纯化的D蛋白制备物体外刺激流感病毒致敏小鼠的脾细胞，选择CD 8 + CTL克隆，促进H1N1和H2 N2病毒的肺清除。在目前的研究中，这些结果通过研究在存在或不存在佐剂的情况下用高度纯化的D蛋白主动免疫的小鼠的应答来扩展。用在氢氧化铝或弗氏完全佐剂中的D蛋白接种CB 6 F1（H-2d x B）小鼠产生针对免疫显性HA 2表位（aa 189-199）的H1N1交叉反应性、H-2d限制性、CD 8 + CTL。不含佐剂的D蛋白不诱导CTL，无论注射途径如何。然而，通过腹腔内（i. p.）在没有佐剂的情况下用D蛋白进行。在第0周和第3周皮下注射含D蛋白的氢氧化铝的小鼠中，在第二次接种后长达16周，相对于对照组，存活率增加，此后需要额外加强免疫以获得保护。在用D蛋白接种的H-2b和H-2k小鼠中的研究表明，在不存在CD 8 + CTL的情况下，诱导CD 4 + T细胞或抗体应答与保护无关。来自CB 6 F1小鼠的免疫血清的被动转移也没有保护性。这种铝佐剂中的原型H1N1重组亚单位疫苗应该直接解决在人流感中实现保护性细胞介导的免疫应答的可行性。
Induction of class I MHC-restricted cytotoxic T lymphocyte (CTL) responses by soluble proteins or peptides requires complex adjuvants or carrier systems which are not licensed for use with human vaccines. The data presented in this report show that vaccination with a highly purified recombinant influenza protein antigen in aluminium hydroxide adjuvant, the only adjuvant currently licensed for clinical use, elicited class I restricted CTL and protection from lethal challenge with H1N1 and H2N2 viruses. The antigen (D protein, SK&F 106160) is produced by expression of H1N1 influenza virus-derived cDNA (strain A/PR/8/34) in Escherichia coli, and is composed of the first 81 N-terminal amino acids (aa) of the non-structural protein 1 (NS1) fused via a nine nucleotide non-viral linker sequence to the 157 C-terminal aa of the haemagglutinin 2 subunit (HA2). Previous work by Kuwano et al1 demonstrated that in vitro stimulation of spleen cells from influenza virus-primed mice, with a partially purified preparation of the D protein, selected for CD8+ CTL clones which facilitated lung clearance of H1N1 and H2N2 viruses. In the current study, these results were extended by studying the responses of mice actively immunized with highly purified D protein in the presence or absence of adjuvants. Vaccination of CB6F1 (H-2d x b) mice with D protein in aluminium hydroxide or Freund's complete adjuvant generated H1N1 cross-reactive, H-2d-restricted, CD8+ CTL directed against an immunodominant HA2 epitope (aa 189-199). D protein without adjuvant did not elicit CTL, regardless of the route of injection. However, long-lived (> 6 months) splenic memory CTL were elicited by boosting mice intraperitoneally (i.p.) with the D protein in the absence of adjuvant. In mice injected subcutaneously with D protein in aluminium hydroxide at weeks 0 and 3, survival was increased relative to controls up to 16 weeks beyond the second vaccination, after which time additional boosting was required for protection. Studies in H-2b and H-2k mice vaccinated with the D protein showed that induction of CD4+ T-cell or antibody responses, in the absence of CD8+ CTL, did not correlate with protection. Passive transfer of immune sera from CB6F1 mice was also not protective. This prototype H1N1 recombinant subunit vaccine in aluminium adjuvant should directly address the feasibility of achieving a protective cell-mediated immune response in human influenza.