Cross-reactivity of monoclonal antibodies against CD4-1 and CD8α of ginbuna crucian carp with lymphocytes of zebrafish and other cyprinid species
Cross-reactivity of monoclonal antibodies against CD4-1 and CD8α of ginbuna crucian carp with lymphocytes of zebrafish and other cyprinid species
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DOI：
10.1016/j.dci.2016.12.002
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发表时间：
2018-03-01
影响因子：
2.9
通讯作者：
Nakanishi, Teruyuki
中科院分区：
文献类型：
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作者：
Miyazawa, Ryuichiro;Matsuura, Yuta;Nakanishi, Teruyuki
我们在银鲫中获得了针对 CD4-1 (6D1) 和 CD8 α (20) 的单克隆抗体 (mAb)。在我们之前的研究中，我们发现 20 mAb 阳性细胞是对同种异体靶点表现出特异性细胞毒性的主要细胞类型，这表明 ginbuna 中的 CD8 α(+) 淋巴细胞相当于哺乳动物中的细胞毒性 T 淋巴细胞 (CTI)。我们通过研究混合白细胞培养物 (MLC) 和半抗原/载体效应，进一步证明了 6D1 mAb 阳性细胞的辅助 T 细胞功能。在这里，我们报告我们的单克隆抗体与斑马鱼淋巴细胞发生交叉反应。首先，mAb 6D1 和 2C3 识别 7-11% 的 ZAP-70 阳性斑马鱼淋巴细胞，并具有典型的淋巴细胞形态。其次，为了验证与 6D1 和 20 mAb 反应的细胞类型，我们检查了来自肾脏的 FACS 分选淋巴细胞中斑马鱼淋巴细胞表面标记的表达谱。在 6D1(2C3)(+) 淋巴细胞中检测到 cd4-1 (cd8 α) 和 tcrac 但未检测到 iglc 转录物，而在 6D1 (2C3)(-) 淋巴细胞中未发现 cd4-1 (cd8 α) 转录物。第三，我们进一步证实6D1与斑马鱼CD4-1反应，但不与CD4-2反应，并且20识别HEK293T细胞上表达的斑马鱼CD8α。总的来说，这些发现表明斑马鱼中的 6D1(+) 和 2C3 淋巴细胞分别相当于哺乳动物中的 CD4(+) 和 CD8 α(+) T 淋巴细胞。此外，我们发现我们的 6D1 和 2C3 mAb 与其他鲤鱼物种（包括金鱼、鲤鱼和草鱼）具有交叉反应性。 (C) 2016 Elsevier Ltd. 保留所有权利。
We have monoclonal antibodies (mAbs) against CD4-1 (6D1) and CD8 alpha (20) in ginbuna crucian carp Carassius auratus langsdorfii. In our previous studies we showed that 20 mAb positive cells are the primary cell type showing specific cytotoxicity against allogeneic targets, suggesting that CD8 alpha(+) lymphocytes in ginbuna are equivalent to cytotoxic T lymphocytes (CTIs) in mammals. We further demonstrated the helper T cell function of 6D1 mAb positive cells by studying mixed leukocyte culture (MLC) and hapten/carrier effects. Here, we report that our mAbs cross-react with zebrafish lymphocytes. First, mAbs 6D1 and 2C3 recognized 7-11% of zebrafish lymphocytes that were ZAP-70 positive and had the typical morphology of lymphocytes. Second, to verify the cell types reacting with the 6D1 and 20 mAbs we examined the expression profiles of zebrafish lymphocyte surface markers in FACS-sorted lymphocytes from kidney. cd4-1 (cd8 alpha) and tcrac but not iglc transcripts were detected in 6D1(2C3)(+) lymphocytes, whereas cd4-1 (cd8 alpha) transcripts were not found in 6D1 (2C3)(-) lymphocytes. Third, we further confirmed that 6D1 reacted with zebrafish CD4-1 but not CD4-2, and 20 recognized zebrafish CD8 alpha expressed on HEK293T cells. Collectively, these findings suggest that 6D1(+) and 2C3 lymphocytes in zebrafish are equivalent to CD4(+) and CD8 alpha(+) T lymphocytes in mammals, respectively. Furthermore, we found the cross-reactivity of our 6D1 and 2C3 mAbs with other cyprinid species including goldfish, common carp and grass carp. (C) 2016 Elsevier Ltd. All rights reserved.