ADJUVANT ENHANCED ANTIVIRAL IMMUNITY

这个子项目是许多利用资源的研究子项目之一 由NIH/NCRR资助的中心拨款提供。子项目的主要支持 而子项目的主要调查员可能是由其他来源提供的， 包括其它NIH来源。 列出的子项目总成本可能 代表子项目使用的中心基础设施的估计数量， 而不是由NCRR赠款提供给子项目或子项目工作人员的直接资金。 甲型流感感染每年在世界范围内引起大量的发病率和死亡率，特别是对于婴儿、老年人和免疫功能低下者。目前的疫苗，例如肠胃外施用的三价灭活疫苗（TIV）是无佐剂的或用氢氧化铝（明矾）佐剂的。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）表面蛋白与当前流行病毒的紧密匹配。如果中和抗体不能预防呼吸道感染，随后病毒感染的清除主要依赖于T细胞，特别是CD 8 + T细胞亚群的溶细胞性T淋巴细胞（CTL）。 目前正在开发一种独特的佐剂，这种佐剂特别有希望用于诱导高水平抗体和T细胞免疫（包括CTL）的疫苗。佐剂基于阳离子脂质载体和非编码DNA复合物（CLDC）。CLDC中包含蛋白抗原导致非常稳健的体液和CD 4+和CD 8 + T细胞应答。CLDC佐剂的独特之处在于，它既作为通过脂质体组分将相关抗原靶向抗原呈递细胞的递送系统，又作为通过质粒DNA的免疫刺激佐剂。该提案产生的最终产品是完成CLDC产品的临床前评价和开发，以用作病毒疫苗（特别是流感疫苗）的佐剂。流感中CLDC佐剂的产品开发策略侧重于利用目前批准的疫苗（在美国和国外）的近期佐剂机会，同时认识到目前正在评估的重组疫苗方法的潜力。本提案的具体目标是：1）确定佐剂和疫苗制备和储存的限度; 2）剂量优化; 3）比较使用现有佐剂或无佐剂接种CLDC/Ag后免疫应答的幅度和持久性; 4）确定佐剂在小鼠和非人灵长类动物中诱导获得性免疫的作用机制; 5）用CLDC佐剂化单价裂解疫苗（其将在内部制备）或单价完整灭活疫苗病毒的异亚型保护程度。有重要的证据表明，异亚型免疫可能遵循自然感染，并在较小程度上，目前的疫苗接种策略。使用CLDC佐剂疫苗可能会显著增强这种免疫力，但在调用这种机制的疫苗策略可以进行临床测试并用于一般应用之前，需要在小鼠和非人灵长类动物中正式证明这一点。

This subproject is one of many research subprojects utilizing the resources provided by a Center grant funded by NIH/NCRR. Primary support for the subproject and the subproject's principal investigator may have been provided by other sources, including other NIH sources. The Total Cost listed for the subproject likely represents the estimated amount of Center infrastructure utilized by the subproject, not direct funding provided by the NCRR grant to the subproject or subproject staff. 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.