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通过⼀个31色深度⾼维的Panel,使用细胞表面染色来检测分型Y细胞亚群和人外周血和淋巴结的免疫表型。除了适应性T细胞标记物外,还包括TCR Vα24-Jα18、TCR γδ、TCR Vɑ7.2和CD161来分别鉴定iNKT、γδ T和MAIT细胞,它们是先天样T细胞。同时纳入了C-X-C趋化因子受体(CXCR3、CXCR4、CXCR5、CXCR6)和C-C基序趋化因子受体(CCR4、CCR6、CCR7),以鉴定Th细胞亚群(Th1、Th2、Th17)、Tfh细胞亚群(Tfh1、Tfh2、Tfh17)和具有特定归巢能力的Th细胞。此外,还使用了标记物来评估细胞分化(CD45RO、CD7)、活化(CD57、CD95、HLA-DR)和⼀些共信号分子(PD-1、NKG2D、CD28)的表达。特别是,CD69和CD103被纳入用于进⼀步分析组织驻留记忆T(Trm)细胞。
标志物 | 荧光染料 | 克隆 | 意义 |
CD45 | APC-R700 | HI30 | White blood cell lineage |
CD3 | BUV805 | UCHT1 | Tcelllineage |
CD4 | BV750 | SK3 | T-celllineage |
CD8 | BV786 | RPA-T8 | Tcelllineage |
CD56 | BV650 | NCAM16.2 | NK cell lineage/NKT-like cel |
CD25 | BV421 | M-A251 | Regulatory Tcell lineage |
CD127 | PE-Cy5 | HI-7R-M21 | Regulatory T-cell lineage |
TCR y7.2 | BV570 | OF-5A12 | MAIT cell lineage |
TCR yó | APC | B1 | y8T cell lineage |
TCR Va24-J18 | BV711 | 6B11 | iNKT cell marker |
CXCR3 | BV510 | 1C6 | Tfh |
CXCR3 | BV510 | CXCR3 | Th marker |
CXCR4 | BUV563 | 12G5 | Th marker |
CXCR5 | BUV615 | RF8B2 | Tfh marker |
CXCR6 | BB700 | 13B 1E5 | Th marker |
CCR4 | BUV395 | 1G1 | Th marker |
CCR6 | BUV496 | 11A9 | Tfh |
CCR6 | BUV496 | 11A9 | Th marker |
CCR7 | APC-Cy7 | 2-L1 ·A | Naive cell marker |
NKG2D | BB660 | 1D11 | Costimulation molecule |
CD69 | BB790 | FN50 | Activation |
CD69 | BB790 | FN50 | tissue residency marker |
CD103 | BB630 | 20000000 | Tissue residency marker |
CD45RO | PE-CF594 | UCHL1 | Memory cell marker |
CD161 | BUV737 | DX12 | MAIT cell marker |
CD28 | PE | CD28.2 | Costimulation molecule |
CD95 | BUV661 | DX2 | Activation |
CD95 | BUV661 | DX2 | exhaustion marker |
CD57 | BV480 | NK-1 | Terminal differentiation |
CD7 | PE-Cy7 | M-T701 | Naïve |
CD7 | PE-Cy7 | M-T701 | effector T cel |
PD-1 | BB515 | M-A251 | Exhaustion marker |
HLA-DR | BV605 | G46-6 | Activation marker |
实验步骤
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结果数据说明
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tSNE analysis of T cells from PBMCs stained using our 31-parameter panel, with main subsets indicated
Cytometry A
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2021 Mar;99(3):273-277. doi: 10.1002/cyto.a.24272. Epub 2020 Dec 10.OMIP 071: A 31-Parameter Flow Cytometry Panel for In-Depth Immunophenotyping of Human T-Cell Subsets Using Surface Markers