Comparative study of immune responses induced after immunization with plasmids encoding the HIV-1 Nef protein under the control of the CMV-IE or the muscle-specific desmin promoter
Comparative study of immune responses induced after immunization with plasmids encoding the HIV-1 Nef protein under the control of the CMV-IE or the muscle-specific desmin promoter
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DOI：
10.1016/s0264-410x(02)00310-9
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发表时间：
2002-09-10
期刊：
影响因子：
5.5
通讯作者：
Bahraoui, E
中科院分区：
文献类型：
--
作者：
Cazeaux, N;Bennasser, Y;Bahraoui, E
本研究的目的是比较DNA免疫后诱导的免疫反应对HIV-1 Nef与Nef编码质粒的控制下的一个普遍存在的或肌肉特异性启动子。为此，构建了含有在巨细胞病毒或人结蛋白启动子控制下的HIV-1 nef的质粒，其在肌细胞中特异性表达。用10 μ g或100 μ g的两种构建体免疫不同组的BALB/c小鼠，对照是pcDNA 3载体或Nef蛋白在弗氏佐剂（Nef-CFA）中。我们的数据表明，这两种质粒刺激抗Nef不道德的反应，在剂量依赖性的方式。然而，与结蛋白启动子相比，CMV-IE启动子的抗Nef抗体应答更早且更高。我们还显示，Nef表达质粒诱导高滴度的抗Nef抗体（104），与Nef-CFA组中获得的那些（4 × 104）相当。抗Nef抗体的特异性分析显示启动子没有影响，并且与Nef-CFA相反，质粒免疫引起主要针对构象依赖性表位的抗Nef抗体。淋巴增生反应的数据表明，表达的特异性，或质粒剂量，不影响发病时间或强度的反应。抗Nef抗体的主要IgG 2a同种型和由Nef刺激的脾细胞产生的细胞因子谱（主要是IL-2和IFN-γ）表明质粒免疫的小鼠中的Th 1应答，与用Nef-CFA免疫的小鼠相反，其中诱导Th 2应答。总之，这些数据表明肌细胞的抗原表达足以刺激Th 1免疫应答。(C)2002爱思唯尔科技有限公司版权所有。
The objective of this study was to compare immune responses induced against HIV-1 Nef after DNA immunization with Nef encoding plasmids under the control of an ubiquitous or a muscle-specific promoter. To this end, plasmids containing HIV-1 nef under the control of the Cytomegalovirus or the human desmin promoters, specifically expressed in muscle cells, were constructed. Different groups of BALB/c mice were immunized with 10 or 100 jig of both constructs, controls were the pcDNA3 vector or the Nef protein in Freund's adjuvant (Nef-CFA). Our data showed that both plasmids stimulated anti-Nef Immoral responses in a dose-dependent manner. The anti-Nef antibody response, however, was earlier and higher with the CMV-IE promoter than with the desmin promoter. We also showed that Nef expressing plasmids induced high titers of anti-Nef antibodies (104), comparable to those obtained in the Nef-CFA group (4 x 104). Analysis of the specificity of anti-Nef antibodies revealed no influence of the promoter, and in contrast to Nef-CFA, plasmid immunization elicited anti-Nef antibodies directed principally against conformational-dependent epitopes. Data on the lymphoproliferative response showed that the specificity of expression, or the plasmid dose, did not affect the onset-time or the intensity of the response. The predominant IgG2a isotype of anti-Nef antibodies and the cytokine profile, mostly IL-2 and IFN-gamma, produced by Nef-stimulated spleen cells indicated a Th1 response in plasmid-immunized mice, in contrast to mice immunized with Nef-CFA, where a Th2 response was induced. In conclusion, these data indicate that antigen expression by muscle cells is sufficient to Stimulate a Th1 immune response. (C) 2002 Elsevier Science Ltd. All rights reserved.