Construction, characterization, and immunogenicity of a multigene modified vaccinia Ankara (MVA) vaccine based on HIV type 1 subtype C
Construction, characterization, and immunogenicity of a multigene modified vaccinia Ankara (MVA) vaccine based on HIV type 1 subtype C
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DOI：
10.1089/aid.2007.0205
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发表时间：
2008-02-01
影响因子：
1.5
通讯作者：
Williamson, Anna-Lise
中科院分区：
文献类型：
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作者：
Burgers, Wendy A.;Shephard, Enid;Williamson, Anna-Lise
候选疫苗由DNA构建体组成，以引发免疫系统，然后是含有匹配基因的改良安卡拉牛痘（MVA）作为加强疫苗接种，在人类志愿者中产生了令人鼓舞的免疫应答。本研究介绍了重组MVA的详细构建和表征，该重组MVA将在南非和美国的I期临床试验中与DNA疫苗组合进行测试。为了匹配最近在南部非洲地区传播的病毒并最大化表位覆盖率，构建的疫苗含有5个HIV-1 C亚型基因，即gag、逆转录酶、达特和nef（grttn），表达为多聚蛋白，以及截短的env（gp 150）。发现含有野生型env的初始重组MVA构建体在遗传上不稳定，因此使用人密码子优化的基因。将Grttn和gp 150插入MVA的两个不同位点，产生双重组体SAAVI MVA-C。重组MVA显示出遗传稳定性，并且观察到转基因的高水平表达。尽管在病毒生成过程中出现了点突变，但Env在功能性感染性测定中保留了感染性。以不同剂量接种SAAVI MVA-C的小鼠产生了高水平的Gag、RT和Env特异性CD 8（+）和CD 4（+）T细胞，其中一些反应可以通过第二次接种得到加强。随附的论文描述了SAAVI MVA-C与SAAVI DNA-C联合给药时的免疫原性。
Candidate vaccines composed of a DNA construct to prime the immune system, followed by modified vaccinia Ankara (MVA) containing matching genes as a booster vaccination, have produced encouraging immune responses in human volunteers. This study presents the detailed construction and characterization of a recombinant MVA that will be tested in combination with a DNA vaccine in Phase I clinical trials in South Africa and the United States. To match recently transmitted viruses in the southern African region and to maximize epitope coverage, the vaccines were constructed to contain five HIV-1 subtype C genes, namely gag, reverse transcriptase, tat, and nef (grttn), expressed as a polyprotein, and a truncated env (gp150). An initial recombinant MVA construct containing wild-type env was found to be genetically unstable, and thus a human codon-optimized gene was used. Grttn and gp150 were inserted into two different sites in MVA yielding a double recombinant, SAAVI MVA-C. The recombinant MVA was shown to be genetically stable and high level expression of the transgenes was observed. Env retained infectivity in a functional infectivity assay despite a point mutation that arose during virus generation. Mice inoculated with SAAVI MVA-C at various doses developed high levels of Gag, RT, and Env-specific CD8(+) and CD4(+) T cells, and some of these responses could be boosted by a second inoculation. An accompanying paper describes the immunogenicity of SAAVI MVA-C when given in combination with SAAVI DNA-C.