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使用流式细胞术对慢性髓系恶性肿瘤患者循环未成熟粒细胞进行定量分析的方案。

Protocol for quantification of circulating immature granulocytes in patients with a chronic myeloid malignancy using flow cytometry.

作者信息

Morabito Margot, Solary Eric, Selimoglu-Buet Dorothée

机构信息

INSERM U1287, Gustave Roussy Cancer Center, Villejuif, France; Université Paris-Saclay, Faculté de Médecine, Le Kremlin-Bicêtre, France.

INSERM U1287, Gustave Roussy Cancer Center, Villejuif, France; Université Paris-Saclay, Faculté de Médecine, Le Kremlin-Bicêtre, France.

出版信息

STAR Protoc. 2025 Aug 4;6(3):104009. doi: 10.1016/j.xpro.2025.104009.

DOI:10.1016/j.xpro.2025.104009
PMID:40763033
原文链接:https://pmc.ncbi.nlm.nih.gov/articles/PMC12345365/
Abstract

Immature granulocytes (iGRANs) circulating in the peripheral blood of patients with a chronic myeloid malignancy, such as chronic myelomonocytic leukemia, are immunosuppressive cells that promote disease progression. Here, we present a multiparameter flow cytometry-based protocol to quantify iGRANs among peripheral blood mononucleated cells (PBMCs). We describe steps for collecting and labeling PBMCs. We describe the gating strategy to quantify iGRANs as a fraction of myeloid cells or as an absolute number of circulating cells, which supports patient stratification and monitoring. For complete details on the use and execution of this protocol, please refer to Deschamps et al..

摘要

在慢性髓系恶性肿瘤患者（如慢性粒单核细胞白血病患者）外周血中循环的未成熟粒细胞（iGRANs）是促进疾病进展的免疫抑制细胞。在此，我们提出一种基于多参数流式细胞术的方案，用于定量外周血单核细胞（PBMCs）中的iGRANs。我们描述了收集和标记PBMCs的步骤。我们描述了将iGRANs定量为髓系细胞一部分或作为循环细胞绝对数量的门控策略，这有助于患者分层和监测。有关本方案使用和执行的完整详细信息，请参考Deschamps等人的文献。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/d5bd/12345365/b7f456f0ba51/gr7.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/d5bd/12345365/c6029c8a4e4a/fx1.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/d5bd/12345365/3fee7eef7410/gr1.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/d5bd/12345365/0f10ff6b4b4c/gr2.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/d5bd/12345365/11661e9044c3/gr3.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/d5bd/12345365/06eee6f42882/gr4.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/d5bd/12345365/b6dd10b84cab/gr5.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/d5bd/12345365/f543d02f2d0f/gr6.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/d5bd/12345365/b7f456f0ba51/gr7.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/d5bd/12345365/c6029c8a4e4a/fx1.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/d5bd/12345365/3fee7eef7410/gr1.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/d5bd/12345365/0f10ff6b4b4c/gr2.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/d5bd/12345365/11661e9044c3/gr3.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/d5bd/12345365/06eee6f42882/gr4.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/d5bd/12345365/b6dd10b84cab/gr5.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/d5bd/12345365/f543d02f2d0f/gr6.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/d5bd/12345365/b7f456f0ba51/gr7.jpg

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本文引用的文献

1
Characterizing a low-density neutrophil gene signature in acute and chronic infections and its impact on disease severity.鉴定急性和慢性感染中的低密度中性粒细胞基因特征及其对疾病严重程度的影响。
J Leukoc Biol. 2025 Apr 23;117(4). doi: 10.1093/jleuko/qiaf027.
2
CXCL8 secreted by immature granulocytes inhibits WT hematopoiesis in chronic myelomonocytic leukemia.不成熟粒细胞分泌的 CXCL8 抑制慢性粒单核细胞白血病中的 WT 造血。
J Clin Invest. 2024 Sep 17;134(22):e180738. doi: 10.1172/JCI180738.
3
Prognostic role of pretreatment circulating MDSCs in patients with solid malignancies: A meta-analysis of 40 studies.
实体恶性肿瘤患者治疗前循环髓源性抑制细胞的预后作用：40项研究的荟萃分析
Oncoimmunology. 2018 Jul 30;7(10):e1494113. doi: 10.1080/2162402X.2018.1494113. eCollection 2018.