Biological activity of human IgE monoclonal antibodies targeting Der p 2, Fel d 1, Ara h 2 in basophil mediator release assays.
Biological activity of human IgE monoclonal antibodies targeting Der p 2, Fel d 1, Ara h 2 in basophil mediator release assays.
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DOI：
10.3389/fimmu.2023.1155613
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发表时间：
2023
影响因子：
7.3
通讯作者：
Aglas, Lorenz
中科院分区：
文献类型：
--
作者：
Pena-Castellanos, Glorismer;Smith, Bryan R. E.;Pomes, Anna;Smith, Scott A.;Stigler, Maria A.;Widauer, Hannah L.;Versteeg, Serge A.;van Ree, Ronald;Chapman, Martin D.;Aglas, Lorenz
人免疫球蛋白E单抗(HIGE MAb)是研究IgE反应的独特工具。本文研究了从变态反应性献血者血液中提取的永生化B细胞产生的针对三种变应原(Der-p-2、Fel-d-1和Ara-h-2)的HIGE单抗的生物活性。由人B细胞杂交瘤产生的3株Der p2-、3株Fel d1-和5株Arah2特异性的hIGE单抗成对结合，用来被动致敏人源化的大鼠嗜碱性白血病细胞，并与血清池致敏进行比较。用相应的过敏原(重组或纯化)、过敏原提取物或结构同源物刺激致敏细胞，序列相似性为40-88%，并比较介质(β-氨基己糖苷酶)的释放。分别有1对、2对和8对Der p2-、Fel d 1-和Ara h 2特异性的hIGE单抗产生显著的介质释放(>；50%)。最低mAb浓度为15-30kU/L，最低抗原浓度为0.01-0.1微克/毫升，足以引起明显的介质释放。用一种Arah2特异性的HIGE单抗单独致敏能够诱导不依赖于第二种特异性HIGE单抗的交联。与同系物相比，DERP2和Arah2特异性mAb表现出更高的过敏原特异性。HIGE单抗致敏的细胞释放的介质与血清致敏的相当。本文报道的HIGE单抗的生物学活性为应用HIGE单抗进行过敏原产品标准化和质量控制的新方法以及对IgE介导的过敏性疾病的机制研究提供了基础。
Human Immunoglobulin E monoclonal antibodies (hIgE mAb) are unique tools for investigating IgE responses. Here, the biological activity of hIgE mAb, derived from immortalized B cells harvested from the blood of allergic donors, targeting three allergens (Der p 2, Fel d 1 and Ara h 2) was investigated. Three Der p 2-, three Fel d 1- and five Ara h 2-specific hIgE mAb produced by human B cell hybridomas, were combined in pairs and used to passively sensitize humanized rat basophilic leukemia cells and compared with sensitization using serum pools. Sensitized cells were stimulated with corresponding allergens (recombinant or purified), allergen extracts or structural homologs, having 40-88% sequence similarity, and compared for mediator (β-hexosaminidase) release. One, two and eight pairs of Der p 2-, Fel d 1- and Ara h 2-specific hIgE mAb, respectively, produced significant mediator release (>50%). A minimum hIgE mAb concentration of 15-30 kU/L and a minimum antigen concentration between 0.01-0.1 µg/mL were sufficient to induce a pronounced mediator release. Individual sensitization with one Ara h 2-specific hIgE mAb was able to induce crosslinking independently of a second specific hIgE mAb. Der p 2- and Ara h 2-specific mAb showed a high allergen specificity when compared to homologs. Mediator release from cells sensitized with hIgE mAb was comparable to serum sensitization. The biological activity of hIgE mAb reported here provides the foundation for novel methods of standardization and quality control of allergen products and for mechanistic studies of IgE-mediated allergic diseases, using hIgE mAb.
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DOI：
10.4049/jimmunol.2000295
发表时间：
2020-10-15
期刊：
Journal of immunology (Baltimore, Md. : 1950)
影响因子：
--
作者：
Mueller GA;Glesner J;Daniel JL;Zhang J;Hyduke N;Richardson CM;DeRose EF;Chapman MD;Peebles RS Jr;A Smith S;Pomés A
通讯作者：
Pomés A
影响因子：
3.4
作者：
Ladics, G. S.;van Bilsen, J. H. M.;Knippels, L. M. J.
通讯作者：
Knippels, L. M. J.
影响因子：
14.2
作者：
Hemmings, Oliver;Du Toit, George;Santos, Alexandra F.
通讯作者：
Santos, Alexandra F.
影响因子：
5.5
作者：
Smith, Scott A.;Chruszcz, Maksymilian;Chapman, Martin D.;Pomes, Anna
通讯作者：
Pomes, Anna
影响因子：
14.2
作者：
Kaul, S;Scheurer, S;Hoffmann, A
通讯作者：
Hoffmann, A