Higgins Deborah, Rodriguez Roberto, Milley Robert, Marshall Jason, Abbate Christi, dela Cruz Tracy, Patton Kathryn, Walker Fiona, Chichester Kristin, Eiden Joseph, Tuck Stephen, Van Nest Gary
Dynavax Technologies Corp., 2929 Seventh Street, Berkeley, CA 94710, USA.
J Allergy Clin Immunol. 2006 Aug;118(2):504-10. doi: 10.1016/j.jaci.2006.05.001. Epub 2006 Jun 27.
Immunostimulatory DNA sequences (ISS) are potent immunomodulators that can drive T(H)1 responses to antigens or allergens. This effect can be dramatically enhanced by direct linkage of ISS to the protein.
Evaluate the effects of the number of ISS bound to the major ragweed allergen Amb a 1 on immunogenicity and allergenicity.
Immunogenicity in mice and allergenicity using PBMC or sera from subjects with ragweed allergy were assayed.
Both antibody induction in vivo and antibody recognition in vitro were highly sensitive to the number of ISSs linked. IgE recognition of Amb a 1 in competitive ELISA or histamine release assays was inhibited by ISS linkage and showed an inverse relationship to the number of ISSs bound. Type and magnitude of antibody induction in mice was also highly dependent on the number of ISS bound. At the highest ISS to protein ratios, antibody induction was very low. Moderate ISS to protein ratios induced high antibody responses in which IgG(2a) generally predominated. Low ISS to protein ratios produced the highest overall antibody responses in which IgG(1) predominated. In contrast, varied ISS to protein ratios did not affect T-cell responses. In both in vivo mouse studies and in vitro human PBMC studies, all ISS to protein ratios evaluated induced similar responses represented by high levels of IFN-gamma and low levels of T(H)2 cytokines.
Controlling the number of ISS bound to a protein allows manipulation of antibody recognition and induction while retaining the potent T(H)1 properties of an ISS-linked protein.
Immunostimulatory DNA sequence-linked Amb a 1 conjugate represents a safe, novel therapeutic approach for treating ragweed allergy.
免疫刺激DNA序列（ISS）是强大的免疫调节剂，可驱动针对抗原或变应原的T(H)1反应。通过将ISS直接与蛋白质连接，这种效应可显著增强。
评估与主要豚草变应原Amb a 1结合的ISS数量对免疫原性和变应原性的影响。
检测小鼠的免疫原性以及使用豚草过敏受试者的外周血单核细胞（PBMC）或血清检测变应原性。
体内抗体诱导和体外抗体识别对连接的ISS数量高度敏感。在竞争性酶联免疫吸附测定（ELISA）或组胺释放试验中，ISS连接可抑制对Amb a 1的IgE识别，且与结合的ISS数量呈负相关。小鼠体内抗体诱导的类型和强度也高度依赖于结合的ISS数量。在ISS与蛋白质比例最高时，抗体诱导非常低。中等的ISS与蛋白质比例诱导产生高抗体反应，其中IgG(2a)通常占主导。低ISS与蛋白质比例产生最高的总体抗体反应，其中IgG(1)占主导。相比之下，不同的ISS与蛋白质比例不影响T细胞反应。在体内小鼠研究和体外人PBMC研究中，所有评估的ISS与蛋白质比例均诱导出相似的反应，表现为高水平的干扰素-γ和低水平的T(H)2细胞因子。
控制与蛋白质结合的ISS数量可在保留ISS连接蛋白强大的T(H)1特性的同时，操控抗体识别和诱导。
免疫刺激DNA序列连接的Amb a 1缀合物代表了一种治疗豚草过敏的安全、新型治疗方法。
