马 CD25 单克隆抗体提高了对马调节性 T 细胞的检测能力

IF 1.4 3区 农林科学 Q4 IMMUNOLOGY Veterinary immunology and immunopathology Pub Date : 2024-06-06 DOI:10.1016/j.vetimm.2024.110790
Bettina Wagner , Susanna Babasyan , Sophie Wilford , Melissa G. Robbin , Amanda M. de Mestre
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引用次数: 0

摘要

CD25 是白细胞介素-2 受体 α 链,表达于不同免疫细胞的细胞表面,常用于调节性 T 细胞(Tregs)的表型分析。CD25 在维持止血和免疫耐受方面起着至关重要的作用,Treg 细胞参与人类疾病和过敏、自身免疫、癌症、慢性炎症等鼠类模型的研究已得到证实。在马身上,交叉反应的抗人 CD25 抗体曾被用于鉴定 Tregs。在这里,我们开发了马 CD25 单克隆抗体(mAbs),并通过流式细胞术比较了它们与抗人 CD25 抗体的染色模式。这两种试剂的比较是在独立实验室进行的两次单独分析中完成的。总体而言,两个实验室使用抗人 CD25 抗体和马 CD25 mAb 15-1 所获得的马外周血淋巴细胞染色模式相似。这两种试剂在用pokeweed有丝分裂原刺激外周血单核细胞(PBMC)后可鉴定出相似的CD4+CD25+和CD4+CD25+FOXP3+百分比。不过,与抗人 CD25 抗体相比,马 CD25 mAb 15-1 能使马 CD25+ 细胞的染色强度更好,并能提高体内 Tregs 和其他 CD25+ 细胞的百分比,以及在无刺激的情况下培养 PBMC 后的百分比。总之,马 CD25 mAb 为马的 Tregs 和 CD25+ 细胞表型分析提供了新的改良试剂。
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Monoclonal antibodies for equine CD25 improve detection of regulatory T cells in horses

CD25, the interleukin-2 receptor α-chain, is expressed on cell surfaces of different immune cells and is commonly used for phenotyping of regulatory T cells (Tregs). CD25 has essential roles in the maintenance of hemostasis and immune tolerance and Treg cell involvement has been shown in human diseases and murine models for allergy, autoimmunity, cancer, chronic inflammation, and many others. In horses, a cross-reactive anti-human CD25 antibody has previously been used for characterizing Tregs. Here, we developed monoclonal antibodies (mAbs) to equine CD25 and compared their staining pattern with the anti-human CD25 antibody by flow cytometry. The comparison of the two reagents was performed by two separate analyses in independent laboratories. Overall, similar staining patterns for equine peripheral blood lymphocytes were obtained with the anti-human CD25 antibody and equine CD25 mAb 15–1 in both laboratories. Both reagents identified comparable CD4+CD25+ and CD4+CD25+FOXP3+ percentages after stimulation of peripheral blood mononuclear cells (PBMC) with pokeweed mitogen. However, when compared to the anti-human CD25 antibody, the equine CD25 mAb 15–1 resulted in a better staining intensity of the equine CD25+ cells and increased the percentages of Tregs and other CD25+ cells ex vivo and after culturing of PBMC without stimulation. In summary, the equine CD25 mAbs provide new, improved reagents for Tregs and CD25+ cell phenotyping in horses.

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来源期刊
CiteScore
3.40
自引率
5.60%
发文量
79
审稿时长
70 days
期刊介绍: The journal reports basic, comparative and clinical immunology as they pertain to the animal species designated here: livestock, poultry, and fish species that are major food animals and companion animals such as cats, dogs, horses and camels, and wildlife species that act as reservoirs for food, companion or human infectious diseases, or as models for human disease. Rodent models of infectious diseases that are of importance in the animal species indicated above,when the disease requires a level of containment that is not readily available for larger animal experimentation (ABSL3), will be considered. Papers on rabbits, lizards, guinea pigs, badgers, armadillos, elephants, antelope, and buffalo will be reviewed if the research advances our fundamental understanding of immunology, or if they act as a reservoir of infectious disease for the primary animal species designated above, or for humans. Manuscripts employing other species will be reviewed if justified as fitting into the categories above. The following topics are appropriate: biology of cells and mechanisms of the immune system, immunochemistry, immunodeficiencies, immunodiagnosis, immunogenetics, immunopathology, immunology of infectious disease and tumors, immunoprophylaxis including vaccine development and delivery, immunological aspects of pregnancy including passive immunity, autoimmuity, neuroimmunology, and transplanatation immunology. Manuscripts that describe new genes and development of tools such as monoclonal antibodies are also of interest when part of a larger biological study. Studies employing extracts or constituents (plant extracts, feed additives or microbiome) must be sufficiently defined to be reproduced in other laboratories and also provide evidence for possible mechanisms and not simply show an effect on the immune system.
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