Summary of workshop findings for porcine B-cell markers
Summary of workshop findings for porcine B-cell markers
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DOI：
10.1016/s0165-2427(01)00279-3
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发表时间：
2001-07-20
影响因子：
1.8
通讯作者：
Bianchi, ATJ
中科院分区：
文献类型：
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作者：
Boersma, WJA;Zwart, RJ;Bianchi, ATJ
基于第三届国际猪 CD 研讨会第一轮分析的聚类组，在本次研讨会 B 细胞部分的第二轮分析中，通过流式细胞术 (FCM) 和免疫组织化学 (IH) 对包括 8 个内部对照在内的 38 种单克隆抗体 (MAb) 进行了分析。本节中的靶标包括外周血淋巴细胞和从成年动物回肠集合淋巴结 (PP)、肠系膜淋巴结 (MLN) 中分离的细胞、新生仔猪的骨髓细胞以及从妊娠第 105 天的胎儿中分离的胸腺细胞。这 38 种 MAb 的免疫组织化学鉴定出四组，其配体与 CD21 共表达，其组织分布与细胞的特异性相一致，包括B 细胞谱系。另一组杂项抗体似乎可以识别其他细胞，其中一些抗体呈阴性。通过将每种感兴趣的抗体与 SWC7、CD21、sIgM 和多克隆兔抗猪免疫球蛋白抗血清 (RaSwIg) 配对来进行双色流式细胞术 (2C-FCM)。抗 CD21 MAb BB6-11C9（第 20 号）和 IAH-CC51（第 19 号）在之前的研讨会中建立，以及交叉反应性抗人 CD21 B-ly4（编号 146），在第一轮 FCM 分析中聚集在一起，并在 IH 中显示出相似的细胞分布。另一个簇由作为 SWC7 特定提交的标准 CC55（编号 55）和 2A10/8（编号 102）形成。第二个 SWC7 标准 2F6/8（编号 100）单独聚类，但 IH 显示出与其他 SWC7 MAb 相同的反应模式。不幸的是，这项工作无法识别任何其他对 B 细胞具有特异性的新簇，因为其他 MAb 的统计聚类无法通过 IH 或后续的双色 FCM 工作证实。然而，我们可以识别具有相似细胞分布的单克隆抗体。交叉反应性抗人 CD40 G28.5（编号 25）和 STH224（编号 153）的配体在非常相似的靶标上表达，MAb 对 JM1H1（编号 139）与 BB6-10A10（编号 142）的配体以及 MAb 对 3F7/11（编号 115）与 1C2F10 的配体类似地表达（第 187 号）。 (C) 2001 Elsevier Science B.V. 保留所有权利。
Based on cluster groups from the first-round analyses of the Third International Swine CD Workshop, 38 monoclonal antibodies (MAbs) including eight internal controls were analysed by flow cytometry (FCM) and immunohistochemistry (IH) in the second-round analysis of the B-cell section of this workshop. Targets in this section included peripheral blood lymphocytes and cells isolated from ileal Peyer's patches (PP), mesenteric lymph nodes (MLN) of adult animals, bone marrow cells from newborn piglets and thymus cells isolated from foetuses at day 105 of gestation.Immunohistochemistry of these 38 MAbs identified four sets, whose ligands were co-expressed with CD21, which showed a tissue distribution compatible with specificity for cells including those of the B-cell lineage. Another group of miscellaneous antibodies appeared to identify other cells, several antibodies were negative. Two-colour flow cytometry (2C-FCM) was carried out by pairing each antibody of interest with antibodies to SWC7, CD21, sIgM and a polyclonal rabbit anti-swine immunoglobulin antiserum (RaSwIg).The anti-CD21 MAb BB6-11C9 (no. 20) and IAH-CC51 (no. 19), established in previous workshops, as well as the cross-reactive anti-human CD21 B-ly4 (no. 146), clustered together in FCM analyses of the first round and showed similar cellular distribution in IH. A further cluster was formed by the standard CC55 (no, 55) and 2A10/8 (no. 102) submitted as SWC7 specific. The second SWC7 standard 2F6/8 (no. 100) clustered separately, but IH showed an identical pattern of reactivity to the other SWC7 MAb.Unfortunately, this work could not identify any other novel clusters with specificity for B-cells, as the statistical clustering of other MAbs could not be substantiated by IH or subsequent two-colour-FCM work. However, we could identify MAb with similar cellular distribution. The ligands for the cross-reactive anti-human CD40 G28.5 (no. 25) and STH224 (no. 153) were expressed on very similar targets, similarly the ligands for the MAb pair JM1H1 (no. 139) with BB6-10A10 (no. 142) and the MAb pair 3F7/11 (no. 115) with 1C2F10 (no. 187). (C) 2001 Elsevier Science B.V. All rights reserved.