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制备和鉴定抗猪 CD103 的单克隆抗体。

Preparation and identification of monoclonal antibodies against porcine CD103.

机构信息

State Key Laboratory of Veterinary Etiological Biology, Lanzhou Veterinary Research Institute, Chinese Academy of Agricultural Sciences, Yanchanghui Chengguan District Lanzhou, 1 Xujiaping, Gansu, 730046, China.

出版信息

Appl Microbiol Biotechnol. 2022 Jun;106(11):4005-4015. doi: 10.1007/s00253-022-11950-x. Epub 2022 May 23.

Abstract

Dendritic cells (DCs) play an important role in activating, regulating, and maintaining the immune response. CD103+ DCs, one of the DC subpopulations, mainly function in the mucosal immune response. They are responsible for capturing and carrying antigens to the relevant lymph nodes to activate the downstream immune responses. However, there is limited available information regarding the function of CD103+ DCs in the porcine mucosal immune response. In this study, two monoclonal antibodies (mAbs) against porcine CD103 were prepared, and their applications were evaluated by enzyme-linked immunosorbent assay (ELISA), indirect immunofluorescence assay (IFA), and flow cytometry. The produced mAbs (7F3 and 9H3) were both IgG1 subtype with κ chains in the light chain. The 7F3 recognizes a linear epitope (PDLRPRAQVYFSDLE) while 9H3 recognizes another linear epitope (QILDEGQVLLGAVGA). The prepared mAbs could be used in vivo to detect the cells expressing CD103 molecules, giving wide applications of both mAbs. In conclusion, this study successfully prepared 2 mAbs against CD103 protein, and they showed applicability in vivo experiments, which will provide the basis for the study of porcine mucosal immunity. KEY POINTS: • Preparation of monoclonal antibodies against porcine CD103 molecule • Analysis of the distribution of CD103 protein on different cells is possible • Exploration of the CD103 DCs function in porcine mucosal immunity is possible.

摘要

树突状细胞 (DCs) 在激活、调节和维持免疫反应方面发挥着重要作用。CD103+ DCs 是 DC 亚群之一,主要在黏膜免疫反应中发挥作用。它们负责捕获和携带抗原到相关的淋巴结,以激活下游的免疫反应。然而,关于 CD103+ DCs 在猪黏膜免疫反应中的功能,信息有限。在本研究中,制备了两种针对猪 CD103 的单克隆抗体 (mAb),并通过酶联免疫吸附试验 (ELISA)、间接免疫荧光试验 (IFA) 和流式细胞术评估了它们的应用。制备的 mAb (7F3 和 9H3) 均为 IgG1 亚型,轻链为 κ 链。7F3 识别线性表位 (PDLRPRAQVYFSDLE),而 9H3 识别另一个线性表位 (QILDEGQVLLGAVGA)。制备的 mAb 可用于体内检测表达 CD103 分子的细胞,为两种 mAb 的广泛应用提供了基础。总之,本研究成功制备了 2 种针对 CD103 蛋白的 mAb,它们在体内实验中具有适用性,为研究猪黏膜免疫提供了基础。 关键点: • 制备针对猪 CD103 分子的单克隆抗体 • 分析 CD103 蛋白在不同细胞上的分布成为可能 • 探索猪黏膜免疫中 CD103 DCs 的功能成为可能。

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