Department of Animal Science, Aarhus University, Tjele, Denmark.
Department of Poultry Science, Tarbiat Modares University, Tehran, Iran.
Cytometry A. 2022 Jan;101(1):45-56. doi: 10.1002/cyto.a.24304. Epub 2021 Jan 27.
A comprehensive analysis of T cell activation markers in chicken is lacking. Kinetics of T cell activation markers (CD25, CD28, CD5, MHC-II, CD44, and CD45) in response to in vitro stimulation of peripheral blood mononuclear cells with concanavalin A (Con A) were evaluated between two chicken lines selected for high and low levels of mannose-binding lectin in serum (L10H and L10L, respectively) by flow cytometry. L10H chickens showed a stronger response to Con A based on the frequency of T cell blasts in both the CD4 and CD8 compartment. The majority of the proliferating CD4 and CD8 T cells expressed CD25. Proliferating T cells were seen both in the CD4 MHC-II and CD8 MHC-II population. For both CD4 and CD8 T cells, frequencies of CD25 and MHC-II T cells were increased 24 h after stimulation. CD28 frequencies were only increased on CD8 T cells 48 h after stimulation. An increase in the relative surface expression based on mean fluorescence intensity (MFI) upon activation was observed for most markers except CD5. For CD4 T cells, CD28 expression increased 24 h after stimulation whereas MHC-II expression increased after 48 h. For CD8 T cells, a tendency toward an increase in CD25 expression was observed. CD28 expression started to increase 24 h after stimulation and only a transient peak in MHC-II expression on CD8 T cells was observed after 24 h. CD44 and CD45 expressed on CD4 and CD8 T cells increased 24-72 h after stimulation. In summary, the frequency of CD25 and MHC-II T cells were shown to be early markers (24 h) for in vitro activation of both CD4 and CD8 T cells. Frequency of CD28 T cells was a later marker (48 h) and only for CD8 T cells. Surface expression of all markers (MFI) increased permanently or transiently upon activation except for CD5.
鸡的 T 细胞活化标志物的全面分析尚缺乏。通过流式细胞术，评估了在体外用刀豆蛋白 A（Con A）刺激外周血单个核细胞后，两种选择高、低血清甘露糖结合凝集素的鸡系（L10H 和 L10L）中 T 细胞活化标志物（CD25、CD28、CD5、MHC-II、CD44 和 CD45）的动力学。基于 CD4 和 CD8 区室中 T 细胞 blast 的频率，L10H 鸡对 Con A 的反应更强。增殖的 CD4 和 CD8 T 细胞大多表达 CD25。在 CD4 MHC-II 和 CD8 MHC-II 群体中均可见增殖的 T 细胞。对于 CD4 和 CD8 T 细胞，刺激后 24 小时 CD25 和 MHC-II T 细胞的频率增加。刺激后 48 小时仅在 CD8 T 细胞上增加 CD28 频率。除 CD5 外，大多数标记物的相对表面表达（基于平均荧光强度（MFI））在激活时增加。对于 CD4 T 细胞，刺激后 24 小时 CD28 表达增加，而 MHC-II 表达在 48 小时后增加。对于 CD8 T 细胞，观察到 CD25 表达增加的趋势。刺激后 24 小时 CD28 表达开始增加，而 CD8 T 细胞中仅观察到 MHC-II 表达的短暂峰值 24 小时后。CD4 和 CD8 T 细胞上表达的 CD28、CD44 和 CD45 在刺激后 24-72 小时增加。总之，CD25 和 MHC-II T 细胞的频率被证明是 CD4 和 CD8 T 细胞体外激活的早期标志物（24 小时）。CD28 T 细胞的频率是一个较晚的标志物（48 小时），仅针对 CD8 T 细胞。除 CD5 外，所有标记物（MFI）的表面表达在激活时永久或短暂增加。
