Immunophenotyping of chicken peripheral blood lymphocyte subpopulations: Individual variability and repeatability
Immunophenotyping of chicken peripheral blood lymphocyte subpopulations: Individual variability and repeatability
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DOI：
10.1016/j.vetimm.2008.05.012
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发表时间：
2008-10-15
影响因子：
1.8
通讯作者：
Marrone, Babetta L.
中科院分区：
文献类型：
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作者：
Fair, Jeanne M.;Taylor-McCabe, Kirsten J.;Marrone, Babetta L.
T细胞淋巴细胞群可基于细胞表面蛋白的表达（其可通过单克隆抗体在外周血中测量）和淋巴细胞亚群的流式细胞术百分比来划分为亚群。为了准确评估鸟类的免疫能力，必须了解鸟类免疫功能和方法的自然变异性。我们的目标是（1）进一步开发流式细胞术，用于估计家禽外周血中的淋巴细胞亚群，（2）估计方法的重复性和变异性，以及（3）估计用于估计淋巴细胞亚群的最佳抗体和细胞标志物组合。这项工作证明了使用流式细胞术测量外周血中使用抗鸡抗体的淋巴细胞亚群的重复性。从外周血中分离的细胞的免疫荧光染色显示，CD 3（+）抗体与血液中平均约12-24%的淋巴细胞反应，这取决于荧光类型。CD 4（+）和CD 8（+）分子分别在血液中淋巴样细胞的4- 31%和1-10%的范围内表达。荧光标记和抗体公司都有助于结果的变异性，应考虑在未来的家禽流式细胞术研究。(c)2008 Elsevier B. V.保留所有权利。
T-cell lymphocyte populations can be delineated into subsets based on expression of cell surface proteins that can be measured in peripheral blood by monoclonal antibodies and flow cytometry percentages of the lymphocyte subpopulations. In order to accurately assess immunocompetence in birds, natural variability in both avian immune function and the methodology must be understood. Our objectives were to (1) further develop flow cytometry for estimating subpopulations of lymphocytes in peripheral blood from poultry, (2) estimate repeatability and variability in the methodology with respect to poultry in a free-range and environmentally diverse situation, and (3) estimate the best antibody and cell marker combination for estimating lymphocyte subpopulations. This work demonstrated the repeatability of using flow cytometry for measurements of peripheral blood in chickens using anti-chicken antibodies for lymphocyte subpopulations. Immunofluorescence, staining of cells isolated from peripheral blood revealed that the CD3(+) antibodies reacted with an average of approximately 12-24% of the lymphoid cells in the blood, depending on the fluorescence type. The CD4(+) and CD8(+) molecules were expressed in a range of 4-31 % and 1-10% of the lymphoid cells in the blood, respectively. Both fluorescence label and antibody company contribute to the variability of results and should be considered in future flow cytometry studies in poultry. (c) 2008 Elsevier B.V. All rights reserved.