Department of Dermatology, Johannes Gutenberg-University Mainz, Mainz, Germany.
Immunology. 2009 Sep;128(1 Suppl):e193-205. doi: 10.1111/j.1365-2567.2008.02947.x. Epub 2008 Sep 17.
Gene gun-mediated biolistic DNA vaccination with beta-galactosidase (betaGal)-encoding plasmid vectors efficiently modulated antigen-induced immune responses in an animal model of type I allergy, including the inhibition of immunoglobulin E (IgE) production. Here we show that CD4(+) as well as CD8(+) T cells from mice biolistically transfected with a plasmid encoding betaGal under the control of the fascin promoter (pFascin-betaGal) are capable of inhibiting betaGal-specific IgE production after adoptive transfer into naïve recipients. Moreover, suppression of IgE production was dependent on interferon (IFN)-gamma. To analyse the modalities of activation of CD4(+) and CD8(+) T cells regarding the localization of antigen synthesis following gene gun-mediated DNA immunization, we used the fascin promoter and the keratin 5 promoter (pK5-betaGal) to direct betaGal production mainly to dendritic cells (DCs) and to keratinocytes, respectively. Gene gun-mediated DNA immunization with each vector induced considerable activation of betaGal-specific CD8(+) cytotoxic T cells. Cytokine production by re-stimulated CD4(+) T cells in draining lymph nodes and immunoglobulin isotype profiles in sera of immunized mice indicated that immunization with pFascin-betaGal induced a T helper type 1 (Th1)-biased immune response, whereas immunization with pK5-betaGal generated a mixed Th1/Th2 immune response. Nevertheless, DNA vaccination with pFascin-betaGal and pK5-betaGal, respectively, efficiently inhibited specific IgE production in the mouse model of type I allergy. In conclusion, our data show that uptake of exogenous antigen produced by keratinocytes and its presentation by untransfected DCs as well as the presentation of antigen synthesized endogenously in DCs represent equivalent pathways for efficient priming of cellular immune responses.
基因枪介导的生物弹道 DNA 疫苗接种用β-半乳糖苷酶（βGal）编码质粒载体在 I 型过敏动物模型中有效调节抗原诱导的免疫应答，包括抑制免疫球蛋白 E（IgE）的产生。在这里，我们显示了在 fascin 启动子（pFascin-βGal）控制下编码βGal 的质粒转染的 CD4+和 CD8+T 细胞能够在过继转移到幼稚受体内后抑制βGal 特异性 IgE 的产生。此外，IgE 产生的抑制依赖于干扰素（IFN）-γ。为了分析 CD4+和 CD8+T 细胞在基因枪介导的 DNA 免疫接种后抗原合成定位方面的激活方式，我们使用 fascin 启动子和角蛋白 5 启动子（pK5-βGal）分别将βGal 产生主要定向于树突状细胞（DC）和角质形成细胞。每个载体的基因枪介导的 DNA 免疫接种均诱导了可观的βGal 特异性 CD8+细胞毒性 T 细胞的激活。刺激后的 CD4+T 细胞在引流淋巴结中的细胞因子产生和免疫小鼠血清中的免疫球蛋白同种型谱表明，pFascin-βGal 免疫诱导了 Th1 偏向的免疫应答，而 pK5-βGal 免疫产生了混合的 Th1/Th2 免疫应答。然而，分别用 pFascin-βGal 和 pK5-βGal 进行 DNA 疫苗接种，可有效抑制 I 型过敏小鼠模型中的特异性 IgE 产生。总之，我们的数据表明，角质形成细胞产生的外源性抗原的摄取及其由未转染的 DC 呈递以及 DC 内源性合成的抗原的呈递代表了有效启动细胞免疫应答的等效途径。
