Byrne K M, Kim H W, Chew B P, Reinhart G A, Hayek M G
Department of Animnal Sciences, Washington State University, Pullman, USA.
Vet Immunol Immunopathol. 2000 Feb 25;73(2):167-82. doi: 10.1016/s0165-2427(99)00163-4.
Flow cytometry is becoming a commonly used technique to characterize a variety of cells. It provides a powerful application to rapidly determine the relative percentages of T-lymphocyte subsets and B-lymphocytes. The effectiveness of its application, however, is dependent on standardization, especially in a clinical setting. Application of flow cytometry to veterinary diagnostics has been limited by the unavailability of reagents and by the unstandardized characterization of normal values using antibodies not commercially available, but typically provided through the generosity of other researchers. This paper presents a standardized gating protocol, and average values and ranges observed for normal canine and feline blood lymphocytes using commercially available antibodies to cell surface markers for CD5, CD3, CD4, CD8, MHC II, and B lymphocytes. The averages for these markers on gated lymphocytes were as follows: Canine CD5 83.3%, Canine CD4 45.0%, Canine CD8 28.8%, Canine MHC II 98.0%, Canine B Cell 12.9%, Canine CD4/CD8 ratio 1.87, Feline T lymphocytes 77.3%, Feline CD4 44.5%, Feline CD8 25.7%, Feline B Cell 24.1%, Feline CD4/CD8 Ratio 1.75. Normal values were also established for a mixed breed group of dogs, and old versus young dogs. This information will provide researchers and clinicians with a standardized protocol for gating, which establishes a basis for comparison between techniques, and a measure of phenotypic percentages for flow cytometry in normal dogs and cats based on this standardization and commercially available antibodies.
流式细胞术正成为一种用于表征多种细胞的常用技术。它为快速确定T淋巴细胞亚群和B淋巴细胞的相对百分比提供了强大的应用。然而，其应用的有效性取决于标准化，尤其是在临床环境中。流式细胞术在兽医诊断中的应用受到试剂可用性的限制，以及使用非商业可用抗体对正常值进行非标准化表征的限制，这些抗体通常是通过其他研究人员的慷慨提供的。本文介绍了一种标准化的设门方案，以及使用针对CD5、CD3、CD4、CD8、MHC II和B淋巴细胞的细胞表面标志物的商业可用抗体，对正常犬猫血液淋巴细胞观察到的平均值和范围。门控淋巴细胞上这些标志物的平均值如下：犬CD5 83.3%，犬CD4 45.0%，犬CD8 28.8%，犬MHC II 98.0%，犬B细胞12.9%，犬CD4/CD8比值1.87，猫T淋巴细胞77.3%，猫CD4 44.5%，猫CD8 25.7%，猫B细胞24.1%，猫CD4/CD8比值1.75。还为一组混种犬以及老年犬与幼年犬建立了正常值。这些信息将为研究人员和临床医生提供一种标准化的设门方案，该方案为技术之间的比较奠定基础，并基于这种标准化和商业可用抗体，为正常犬猫的流式细胞术提供表型百分比的测量方法。
