Platt Ratree, Ng Terry, Glover Sherry, Roof Michael, Kimura Kayoko, Roth James A
Department of Veterinary Microbiology and Preventive Medicine, College of Veterinary Medicine, Iowa State University, Ames, Iowa 50011, USA.
Boehringer Ingelheim Vetmedica, Inc., USA.
Anal Quant Cytopathol Histpathol. 2013 Aug;35(4):197-204.
To characterize baseline canine lymphocyte phenotypes including lymphocytes coexpressing multiple markers by novel 7-color multiparameter flow cytometry.
Fresh canine peripheral blood lymphocytes of 79 healthy 26-week-old Beagle or Beagle-mix dogs were stained and analyzed.
The high number of samples and acquired flow data (averaging 1.9 x 10(5) cells/sample) allowed the detection of minor lymphocyte subsets coexpressing multiple lymphocyte markers. The averaged percentages of major lymphocyte subsets of CD3+, CD4+, CD8+, CD21+ and gammadelta TCR+ cells from this study were 74.0, 43.6, 14.3, 9.6, and 0.2, respectively, which were comparable but uniquely different from other reports as they were simultaneously detected in the same sample. We demonstrated that the commonly used CD21 and CD3 monoclonal antibody (mAb) clones, previously recommended not to be used in the same staining, could and should be used together with the proper steps of lymphocyte gating. We found a high percentage (10.3%) of unidentified CD21- CD3+ CD4- CD8-gammadelta TCR- lymphocyte subset that has never been reported. The intensive gating strategy and the mean percentages of each lymphocyte subset to their parent subsets and to the total lymphocyte population are presented and discussed.
The canine lymphocyte phenotypes were fully characterized. This novel multiparameter flow cytometry method is a powerful approach to in-crease the accuracy of lymphocyte phenotyping in dogs.
通过新型7色多参数流式细胞术对犬淋巴细胞基线表型进行特征分析，包括共表达多种标志物的淋巴细胞。
对79只健康的26周龄比格犬或比格犬混种犬的新鲜外周血淋巴细胞进行染色和分析。
大量的样本和获取的流式细胞数据（平均每个样本1.9×10⁵个细胞）使得能够检测到共表达多种淋巴细胞标志物的次要淋巴细胞亚群。本研究中CD3⁺、CD4⁺、CD8⁺、CD21⁺和γδTCR⁺细胞等主要淋巴细胞亚群的平均百分比分别为74.0%、43.6%、14.3%、9.6%和0.2%，这些数据与其他报告中的数据具有可比性，但又独特不同，因为它们是在同一样本中同时检测到的。我们证明，以前建议不要在同一染色中使用的常用CD21和CD3单克隆抗体（mAb）克隆，可以并且应该通过适当的淋巴细胞设门步骤一起使用。我们发现了一个从未被报道过的未识别的CD21⁻CD3⁺CD4⁻CD8⁻γδTCR⁻淋巴细胞亚群，其比例较高（10.3%）。本文展示并讨论了密集设门策略以及每个淋巴细胞亚群相对于其母亚群和总淋巴细胞群体的平均百分比。
犬淋巴细胞表型得到了全面表征。这种新型多参数流式细胞术方法是提高犬淋巴细胞表型分析准确性的有力手段。
