ANTIMALARIA VACCINE BASED ON AN INFLUENZA VIRUS VECTOR

我们已经证明，表达外源表位的活流感病毒可以 诱导有效抗体和细胞毒性T细胞（CTL）应答。 具体来说，我们产生了表达CTL的重组流感病毒， 和约氏疟原虫环子孢子（CS）蛋白的B细胞表位。 用这些重组流感病毒单独或 随后是表达整个CS蛋白的重组牛痘病毒， 诱导了针对鼠疟疾的保护性免疫。 我们现建议 扩展这一方法，并将其扩展到由P. 恶性疟原虫。 这项工作应产生必要的数据， 未来用作疫苗的重组病毒的最佳设计 对抗疟疾，也可能对抗其他传染病。 为 为此，我们计划： 1.构建表达B和T细胞的重组流感病毒 人疟原虫恶性疟原虫CS蛋白的表位。 2.确定这些构建体在小鼠中的减毒程度，以及 描述暴露于以下物质的动物的流感特异性免疫应答 病毒载体 3.鉴定体液和T细胞介导的抗疟疾免疫 用重组流感-CS病毒免疫的小鼠的应答。 4.构建另外的流感病毒，其表达以下的选定序列： 第二种恶性疟原虫抗原，Trombospondin相关匿名者 蛋白（TRAP），并评估其在小鼠中的免疫原性。还定义了 针对表达CS和TRAP表位两者的流感病毒的免疫应答。 5.尝试通过引发和加强来增强这些免疫应答 用两种亚型流感病毒的转染子，或两种完全 不同的病毒载体，都表达相同的外源表位。

We have shown that live influenza viruses expressing foreign epitopes can induce an efficient antibody and cytotoxic T cell (CTL) response. Specifically, we generated recombinant influenza viruses expressing CTL and B cell epitopes of the circumsporozoite (CS) protein of P. yoelii. Immunization of mice with these recombinant influenza viruses, alone or followed by a recombinant vaccinia virus expressing the entire CS protein, induced protective immunity against this murine malaria. We now propose to expand this approach and extend it to human malaria caused by P. falciparum. This work should generate data necessary for defining the optimal design of recombinant viruses for their future use as vaccines against malaria and possibly also against other infectious diseases. For this purpose we plan to: 1. Construct recombinant influenza viruses, expressing B and T cell epitopes of the CS protein of the human malaria parasite, P. falciparum. 2. Determine the degree of attenuation of these constructs in mice, and characterize the influenza-specific immune responses of animals exposed to the viral vectors. 3. Characterize the humoral and T cell mediated anti-malaria immune responses of mice immunized with recombinant influenza-CS viruses. 4. Construct additional influenza viruses expressing selected sequences of a second P. falciparum antigen, the Trombospondin Related Anonymous Protein (TRAP), and assess their immunogenicity in mice. Also define the immune response to influenza viruses expressing both CS and TRAP epitopes. 5. Attempt to potentiate these immune responses by priming and boosting with transfectants of two subtypes of influenza viruses, or two entirely different viral vectors, both expressing the same foreign epitopes.