Horner A A, Widhopf G F, Burger J A, Takabayashi K, Cinman N, Ronaghy A, Spiegelberg H L, Raz E
Department of Medicine, The Sam and Rose Stein Institute for Aging, University of California, San Diego, La Jolla 92093-0663, USA.
J Allergy Clin Immunol. 2001 Sep;108(3):417-23. doi: 10.1067/mai.2001.117795.
Immunostimulatory sequence oligodeoxynucleotide (ISS-ODN) is a potent antiallergic immunomodulating agent in mice. However, few studies have addressed its antiallergic potential in human subjects.
We sought to determine whether a phosphoro-thioate ISS-ODN could inhibit IL-4-dependent IgE synthesis by human B cells.
Initially, nonatopic- and atopic-donor PBMCs were incubated with ISS-ODN or mutated oligodeoxynucleotide, and cytokine production and B-cell expression of IFN-gamma receptor and IL-4 receptor were measured by using ELISA and flow cytometry, respectively. In subsequent studies atopic-donor PBMCs were incubated with IL-4 alone or with ISS-ODN or mutated oligodeoxynucleotide. After 14 days, IgE production and IgM, IgG, and IgA production were determined by using ELISA. In select IgE studies cytokines were neutralized with mAbs.
ISS-ODN induced IL-12, IFN-alpha, IFN-gamma, IL-10, and IL-6 production from both nonatopic- and atopic-donor PBMCs. ISS-ODN also increased IFN-gamma receptor and inhibited IL-4 receptor expression on B cells from both donor populations. Furthermore, ISS-ODN inhibited IL-4-dependent IgE production by atopic-donor PBMCs. Neutralization of IL-12, IFN-alpha, IFN-gamma, and IL-10, but not IL-6, attenuated the inhibitory activity of ISS-ODN on IgE production. In contrast to its inhibition of IgE synthesis, ISS-ODN stimulated the production of IgM, IgG, and IgA.
These in vitro studies demonstrate that phos-phorothioate ISS-ODN elicits an innate immune response by PBMCs, which inhibits IL-4-dependent IgE synthesis. In addition, these results provide further support for consideration of ISS-ODN therapy for the treatment of allergic disease in clinical practice.
免疫刺激序列寡脱氧核苷酸（ISS-ODN）是一种在小鼠中有效的抗过敏免疫调节剂。然而，很少有研究探讨其在人类受试者中的抗过敏潜力。
我们试图确定硫代磷酸酯ISS-ODN是否能抑制人B细胞中白细胞介素-4（IL-4）依赖性IgE的合成。
首先，将非特应性和特应性供体的外周血单个核细胞（PBMC）与ISS-ODN或突变的寡脱氧核苷酸一起孵育，分别使用酶联免疫吸附测定（ELISA）和流式细胞术检测细胞因子的产生以及B细胞上γ干扰素受体（IFN-γ受体）和IL-4受体的表达。在随后的研究中，将特应性供体的PBMC单独与IL-4或与ISS-ODN或突变的寡脱氧核苷酸一起孵育。14天后，使用ELISA测定IgE的产生以及IgM、IgG和IgA的产生。在选定的IgE研究中，用单克隆抗体（mAb）中和细胞因子。
ISS-ODN诱导非特应性和特应性供体PBMC产生IL-12、α干扰素（IFN-α）、IFN-γ、IL-10和IL-6。ISS-ODN还增加了两个供体群体B细胞上IFN-γ受体的表达并抑制了IL-4受体的表达。此外，ISS-ODN抑制特应性供体PBMC产生IL-4依赖性IgE。中和IL-12、IFN-α、IFN-γ和IL-10，但不包括IL-6，会减弱ISS-ODN对IgE产生的抑制活性。与其对IgE合成的抑制作用相反，ISS-ODN刺激了IgM、IgG和IgA的产生。
这些体外研究表明，硫代磷酸酯ISS-ODN引发PBMC的天然免疫反应，从而抑制IL-4依赖性IgE的合成。此外，这些结果为在临床实践中考虑使用ISS-ODN治疗过敏性疾病提供了进一步的支持。
