It has become anticipated that regenerative medicine will extend into the field of veterinary medicine as new treatments for various disorders. Although the use of allogeneic stem cells for tissue regeneration is more attractive than that of autologous cells in emergencies, the therapeutic potential of allogeneic transplantation is often limited by allo-immune responses inducing graft rejection. Therefore, a methodology for quantifying and monitoring alloreactive T cells is necessary for evaluating allo-immune responses. The mixed lymphocyte reaction (MLR) is widely used to evaluate T cell alloreactivity. In human, flow cytometric MLR with carboxyfluorescein diacetate succinimidyl ester has been established and used as a more useful assay than conventional MLR with radioisotope labeling. However, the available information about alloreactivity based on the differences of dog major histocompatibility complex (MHC) (dog leukocyte antigen, DLA) is quite limited in dog. In this paper, we describe our established flow cytometric MLR method that can quantify the T cell alloreactivity while distinguishing cell phenotypes in dog, and T cell alloreactivity among DLA-type matched pairs was significantly lower than DLA-mismatched pairs, suggesting that our developed flow cytometric MLR method is useful for quantifying T cell alloreactivity. In addition, we demonstrated the advantage of DLA homozygous cells as a donor (stimulator) for allogeneic transplantation. We also elucidated that the frequency of alloreactive T cell precursors was almost the same as that of mouse and human (1-10%). To our knowledge, this is the first report to focus on the degree of allo-immune responses in dog based on the differences of DLA polymorphisms.

中文翻译：

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使用流式细胞仪混合淋巴细胞反应法在犬中根据MHC不相容程度评估同种反应性T细胞。

可以预见的是，再生医学将作为各种疾病的新疗法而扩展到兽医学领域。尽管在紧急情况下使用同种异体干细胞进行组织再生比自体干细胞更具吸引力，但同种异体移植的治疗潜力通常受到诱导移植排斥的同种免疫反应的限制。因此，定量和监测同种反应性T细胞的方法对于评估同种免疫反应是必要的。混合淋巴细胞反应（MLR）被广泛用于评估T细胞同种异体反应性。在人类中，已经建立了具有羧基荧光素二乙酸琥珀酰亚胺酯的流式细胞仪MLR，并且该方法比具有放射性同位素标记的常规MLR更为有用。然而，基于狗主要组织相容性复合物（MHC）（狗白细胞抗原，DLA）的差异，有关变态反应的可用信息在狗中非常有限。在本文中，我们描述了已建立的流式细胞仪MLR方法，该方法可以在区分狗的细胞表型的同时量化T细胞同种异体反应性，并且DLA型匹配对之间的T细胞同种异体性明显低于DLA不匹配对。细胞计数MLR方法可用于定量T细胞同种异体反应。此外，我们证明了DLA纯合细胞作为同种异体移植的供体（刺激物）的优势。我们还阐明了同种反应性T细胞前体的频率与小鼠和人类的频率几乎相同（1-10％）。据我们所知，