Protocol to quantify and phenotype SARS-CoV-2-specific T cell response using a rapid flow-cytometry-based whole blood assay.
Protocol to quantify and phenotype SARS-CoV-2-specific T cell response using a rapid flow-cytometry-based whole blood assay.
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DOI：
10.1016/j.xpro.2022.101771
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发表时间：
2022-12-16
期刊：
影响因子：
--
通讯作者：
Riou, Catherine
中科院分区：
文献类型：
--
作者：
Mutavhatsindi, Hygon;Riou, Catherine
监测抗原特异性T细胞的频率、功能和表型对于评估宿主对病原体或新型疫苗的免疫反应至关重要。在这里，我们描述了一种快速而简单的体外全血检测方法，用于检测SARS-CoV-2特异性T细胞反应并进行表型鉴定。我们详细介绍了用SARS-CoV-2刺激肽刺激全血以及随后的细胞固定和冷冻保存的步骤。我们进一步描述了用于流式细胞术分析的解冻和细胞染色步骤。这种方法最大限度地减少了样本操作，并具有快速的周转时间。有关本议定书的使用和执行的完整细节，请参阅。全血中抗原特异性T细胞的检测和表型分析细胞固定、冷冻、解冻和流式细胞仪分析方法的详细步骤需要最小的血量并保留生理条件方案具有快速周转时间和最大限度地减少样本操纵出版者注：进行任何实验方案都需要遵守当地机构关于实验室安全和伦理的指导方针。监测抗原特异性T细胞的频率、功能和表型对于评估宿主对病原体或新型疫苗的免疫反应至关重要。在这里，我们描述了一种快速而简单的体外全血检测方法，用于检测SARS-CoV-2特异性T细胞反应并进行表型鉴定。我们详细介绍了用SARS-CoV-2刺激肽刺激全血以及随后的细胞固定和冷冻保存的步骤。我们进一步描述了用于流式细胞术分析的解冻和细胞染色步骤。这种方法最大限度地减少了样本操作，并具有快速的周转时间。
Monitoring antigen-specific T cell frequency, function, and phenotype is essential to assess the host immune response to pathogens or novel vaccines. Here, we describe a rapid and simple ex vivo whole blood assay to detect and phenotype the SARS-CoV-2-specific T cell response. We detail steps for whole blood stimulation with SARS-CoV-2 spike peptide and subsequent cell fixation and cryopreservation. We further describe thawing and cell staining steps for flow cytometry analysis. This approach minimizes sample manipulation and has a quick turnaround time. For complete details on the use and execution of this protocol, please refer to. Detection and phenotyping of antigen-specific T cells from whole blood Detailed steps for cell fixation, cryopreservation, thawing, and flow cytometry analysis Approach requires minimal blood volume and preserves physiological conditions Protocol with a quick turnaround time and minimized sample manipulation Publisher’s note: Undertaking any experimental protocol requires adherence to local institutional guidelines for laboratory safety and ethics. Monitoring antigen-specific T cell frequency, function, and phenotype is essential to assess the host immune response to pathogens or novel vaccines. Here, we describe a rapid and simple ex vivo whole blood assay to detect and phenotype the SARS-CoV-2-specific T cell response. We detail steps for whole blood stimulation with SARS-CoV-2 spike peptide and subsequent cell fixation and cryopreservation. We further describe thawing and cell staining steps for flow cytometry analysis. This approach minimizes sample manipulation and has a quick turnaround time.
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影响因子：
15.9
作者：
Riou, Catherine;du Bruyn, Elsa;Wilkinson, Robert J.
通讯作者：
Wilkinson, Robert J.
影响因子：
7.3
作者：
Hardy MY;Goel G;Russell AK;Chen Yi Mei SLG;Brown GJE;Wang S;Szymczak E;Zhang R;Goldstein KE;Neff KM;Williams LJ;Truitt KE;Dzuris JL;Tye-Din JA;Anderson RP
通讯作者：
Anderson RP
影响因子：
2.2
作者：
Horton, Helen;Thomas, Evan P.;De Rosa, Stephen C.
通讯作者：
De Rosa, Stephen C.
影响因子：
5.8
作者：
Riou C;Du Bruyn E;Ruzive S;Goliath RT;Lindestam Arlehamn CS;Sette A;Sher A;Barber DL;Wilkinson RJ
通讯作者：
Wilkinson RJ
影响因子：
2.2
作者：
Hanekom, WA;Hughes, J;Kaplan, G
通讯作者：
Kaplan, G