Induction of a protective response in mice by the dengue virus NS3 protein using DNA vaccines.
Induction of a protective response in mice by the dengue virus NS3 protein using DNA vaccines.
复制标题
DOI：
10.1371/journal.pone.0025685
复制
发表时间：
2011
期刊：
影响因子：
3.7
通讯作者：
Alves AM
中科院分区：
文献类型：
--
作者：
Costa SM;Yorio AP;Gonçalves AJ;Vidale MM;Costa EC;Mohana-Borges R;Motta MA;Freire MS;Alves AM
登革热非结构蛋白3(NS3)是一种多功能蛋白，N-末端含有丝氨酸蛋白酶结构域，C-末端含有解旋酶、NTPase和RTPase结构域。这种蛋白被认为是登革热感染期间CD4+和CD8+T细胞反应的主要靶点，可能参与保护作用。然而，很少有研究评估这种蛋白作为登革热和其他黄病毒的保护性抗原的用途。在目前的工作中，我们研究了基于DENV2的NS3蛋白的DNA疫苗的保护效果。构建了不同的重组质粒，编码全长NS3蛋白或仅编码其功能结构域(蛋白酶和解旋酶)，与信号肽(t-PA)融合或不融合。重组蛋白在BHK-21细胞中成功表达，只有编码t-PA信号序列的重组蛋白才能介导蛋白分泌。用不同的DNA疫苗免疫BALB/c小鼠，并用致死剂量的DENV2攻击。用编码全长NS3或解旋酶结构域的质粒免疫的大多数动物都能在攻击中存活下来，无论t-PA是否存在。然而，一些小鼠表现出高发病率的临床感染症状(后腿瘫痪和驼背姿势)，主要是在基于解旋酶结构域的DNA疫苗免疫的动物组中。另一方面，接种编码蛋白酶域的质粒并不能产生任何保护作用，因为这些小鼠组的死亡率和发病率与在对照组动物中检测到的相似。用特异性CD8+T细胞NS3多肽ELISPOT分析细胞免疫应答。结果显示，以全长蛋白为基础的DNA疫苗诱导了干扰素-γ的产生，从而提示免疫系统的这一分支参与了保护作用。
The dengue non-structural 3 (NS3) is a multifunctional protein, containing a serino-protease domain, located at the N-terminal portion, and helicase, NTPase and RTPase domains present in the C-terminal region. This protein is considered the main target for CD4+ and CD8+ T cell responses during dengue infection, which may be involved in protection. However, few studies have been undertaken evaluating the use of this protein as a protective antigen against dengue, as well as other flavivirus. In the present work, we investigate the protective efficacy of DNA vaccines based on the NS3 protein from DENV2. Different recombinant plasmids were constructed, encoding either the full-length NS3 protein or only its functional domains (protease and helicase), fused or not to a signal peptide (t-PA). The recombinant proteins were successfully expressed in transfected BHK-21 cells, and only plasmids encoding the t-PA signal sequence mediated protein secretion. Balb/c mice were immunized with the different DNA vaccines and challenged with a lethal dose of DENV2. Most animals immunized with plasmids encoding the full-length NS3 or the helicase domain survived challenge, regardless of the presence of the t-PA. However, some mice presented clinical signs of infection with high morbidity (hind leg paralysis and hunched posture), mainly in animal groups immunized with the DNA vaccines based on the helicase domain. On the other hand, inoculation with plasmids encoding the protease domain did not induce any protection, since mortality and morbidity rates in these mouse groups were similar to those detected in the control animals. The cellular immune response was analyzed by ELISPOT with a specific-CD8+ T cell NS3 peptide. Results revealed that the DNA vaccines based on the full-length protein induced the production of INF-γ, thus suggesting the involvement of this branch of the immune system in the protection.
登录
查看更多内容
影响因子：
6.7
作者：
Balsitis SJ;Williams KL;Lachica R;Flores D;Kyle JL;Mehlhop E;Johnson S;Diamond MS;Beatty PR;Harris E
通讯作者：
Harris E
影响因子：
64.8
作者：
HALSTEAD, SB;OROURKE, EJ
通讯作者：
OROURKE, EJ
DOI：
10.1016/s0928-0197(98)00039-7
发表时间：
1998-07-15
期刊：
CLINICAL AND DIAGNOSTIC VIROLOGY
影响因子：
--
作者：
Brinton, MA;Kurane, I;Ennis, FA
通讯作者：
Ennis, FA
影响因子：
3.7
作者：
Bartelma, G;Padmanabhan, R
通讯作者：
Padmanabhan, R
影响因子：
3.7
作者：
Cui, TA;Sugrue, RJ;Fu, JL
通讯作者：
Fu, JL