ADJUVANT ENHANCED ANTIVIRAL IMMUNITY

这个子项目是许多研究子项目中利用 资源由NIH/NCRR资助的中心拨款提供。子项目和 调查员(PI)可能从NIH的另一个来源获得了主要资金， 并因此可以在其他清晰的条目中表示。列出的机构是 该中心不一定是调查人员的机构。 甲型流感感染每年在世界范围内造成相当大的发病率和死亡率，特别是对婴儿、老年人和免疫缺陷者。目前的疫苗，如非肠道注射的三价灭活疫苗(TIV)，要么不加佐剂，要么添加氢氧化铝(明矾)佐剂。这些疫苗的效力高度依赖于疫苗的血凝素(HA)和神经氨酸酶(NA)表面蛋白与目前流行的病毒的紧密匹配。如果中和抗体不能预防呼吸道感染，病毒感染的清除主要依靠T细胞，特别是CD8+T细胞亚群的细胞溶解T淋巴细胞(CTL)。一种独特的佐剂正在开发中，这种佐剂特别适用于同时诱导高水平抗体和T细胞免疫的疫苗，包括CTL。该佐剂以阳离子脂质载体和非编码DNA复合体(CLDC)为基础。将蛋白质抗原与CLDC结合，可产生非常强大的体液反应以及CD4+和CD8+T细胞反应。CLDC佐剂的独特之处在于，它既是一个通过脂质体将相关抗原靶向抗原提呈细胞的递送系统，也是通过质粒DNA作为免疫刺激佐剂的功能。这项提议产生的最终产品是完成CLDC产品的临床前评估和开发，用作病毒疫苗的佐剂，特别是用于流感。用于流感的CLDC佐剂的产品开发战略侧重于开发对(美国和国外)目前批准的疫苗进行佐剂的近期机会，同时认识到目前正在评估的重组疫苗方法的潜力。该建议的具体目标是：1)建立佐剂和疫苗制备和储存的限制；2)优化剂量；3)比较有佐剂和无佐剂的CLDC/Ag疫苗接种后免疫应答的强度和持久性；4)确定佐剂在诱导小鼠和非人灵长类动物适应性免疫中的作用机制；5)将由内部配制的CLDC佐剂单价裂解疫苗或单价全灭活疫苗对异亚型的保护程度。有重要证据表明，自然感染之后可能会出现异亚型免疫，在较小程度上，也可能是当前的疫苗接种策略。使用CLDC佐剂疫苗可能会显著增强这种免疫力，但需要在小鼠和非人类灵长类动物中正式证明这一点，然后才能对调用这种机制的疫苗策略进行临床测试并广泛应用。

This subproject is one of many research subprojects utilizing the resources provided by a Center grant funded by NIH/NCRR. The subproject and investigator (PI) may have received primary funding from another NIH source, and thus could be represented in other CRISP entries. The institution listed is for the Center, which is not necessarily the institution for the investigator. Influenza A infection causes annual substantial morbidity and mortality worldwide, particularly for infants, the elderly, and the immuncompromised. Current vaccines, such as the parenterally administered trivalent inactivated vaccine (TIV) are administered either unadjuvanted or adjuvanted with aluminum hydroxide (alum). The efficacy of these vaccines is highly dependent on close matching of the hemagglutinin (HA) and neuraminidase (NA) surface proteins of the vaccine with currently circulating virus. Should neutralizing antibody fail to prevent infection of the respiratory tract, subsequent clearance of viral infection is mainly dependent on T cells, particularly cytolytic T lymphocytes (CTL) of the CD8+ T-cell subset. A unique adjuvant that is particularly promising for vaccines that induce both high levels of antibody and T-cell immunity, including CTL is being developed. The adjuvant is based on a cationic lipid carrier and non-coding DNA complex (CLDC). Inclusion of protein antigens with CLDC results in an extremely robust humoral and CD4+ and CD8+ T-cell response. The unique aspect of the CLDC adjuvant is that it functions both as a delivery system that targets associated antigens to antigen presenting cells via the liposome component and an immunostimulatory adjuvant via the plasmid DNA. The final product resulting from this proposal is the completion of pre-clinical evaluation and development of the CLDC product for use as an adjuvant in viral vaccines and specifically for influenza. The product development strategy for the CLDC adjuvant in influenza is focused on the exploitation of the near-term opportunity of adjuvanting currently approved vaccines (in the U.S. and abroad) while recognizing the potential of recombinant vaccine approaches currently being evaluated. The specific goals of this proposal are to: 1) Establish limits on adjuvant and vaccine preparation and storage; 2) Optimization of dose; 3) Comparison of the magnitude and durability of the immune response following CLDC/Ag vaccination with existing adjuvants or no adjuvants; 4) Determine mechanism of action of the adjuvant in inducing adaptive immunity in mouse and non-human primate; 5) extent of heterosubtypic protection with CLDC adjuvanted monovalent split vaccines, which will be prepared in house, or monovalent whole inactivated vaccine virus. There is significant evidence that heterosubtypic immunity may follow natural infection and, to a lesser extent, current vaccination strategies. Such immunity will likely be markedly enhanced using CLDC adjuvanted vaccines, but a formal demonstration of this in mice and non-human primates is required before vaccine strategies invoking this mechanism can be clinically tested and adopted for general application.