用于人类外周血单核细胞的 38 色高维免疫分型面板。

IF 1.6 4区医学 Q4 BIOCHEMICAL RESEARCH METHODS Journal of immunological methods Pub Date : 2024-07-09 DOI:10.1016/j.jim.2024.113726

Jeremy Anderson , Leanne Quah , Kiara Mangano , Daniel G. Pellicci , Nadia Mazarakis , Paul V. Licciardi

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引用次数: 0

摘要

高维免疫表型板是对外周血单核细胞（PBMC）进行广泛表型的宝贵资源。我们设计了 38 种颜色的高维表型检测板，利用 5 激光 Cytek Aurora 检测先天性细胞（单核细胞、树突状细胞、NK 细胞、嗜碱性粒细胞、先天性类淋巴细胞）、T 细胞（γδ T 细胞、MAIT 细胞、CD4 和 CD8 记忆细胞、Th1 细胞、Th2 细胞、Th17 细胞、Tfh 细胞、Treg 细胞）和 B 细胞（记忆细胞、浆细胞、过渡性 B 细胞、浆细胞、IgG 细胞、IgM 细胞）亚群及其活化状态。我们优化了最佳荧光色素组合和滴度，以尽量减少罕见免疫细胞群的扩散和自发荧光，并在 15 名健康成人的 PBMCs 上测试了这一面板。这种高维面板对于直接进行体内外研究以评估人体健康和疾病背景下的免疫细胞非常有价值，尤其是在样本有限的情况下。

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A 38-colour high dimensional immunophenotyping panel for human peripheral blood mononuclear cells

High dimensional immunophenotyping panels are invaluable resources for performing extensive phenotyping on peripheral blood mononuclear cells (PBMCs). We designed a 38-colour high dimensional phenotyping panel to measure innate (monocytes, dendritic cells, NK cells, basophils, innate like lymphoid cells), T cell (γδ T cells, MAIT cells, CD4 and CD8 memory, Th1, Th2, Th17, Tfh, Treg) and B cell (memory, plasma cells, transitional B cells, plasmablasts, IgG, IgM) subsets in addition to their activation status using the 5-laser Cytek Aurora. We optimised optimal fluorochrome combinations and titres to minimise spread and autofluorescence of rare immune cell populations and tested this panel on PBMCs from 15 healthy adults. This high dimensional panel will be invaluable for direct ex vivo studies to evaluate immune cells in the context of human health and disease, especially when samples are limited.

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来源期刊

Journal of immunological methods 医学-免疫学

CiteScore

4.10

自引率

0.00%

发文量

120

审稿时长

3 months

期刊介绍： The Journal of Immunological Methods is devoted to covering techniques for: (1) Quantitating and detecting antibodies and/or antigens. (2) Purifying immunoglobulins, lymphokines and other molecules of the immune system. (3) Isolating antigens and other substances important in immunological processes. (4) Labelling antigens and antibodies. (5) Localizing antigens and/or antibodies in tissues and cells. (6) Detecting, and fractionating immunocompetent cells. (7) Assaying for cellular immunity. (8) Documenting cell-cell interactions. (9) Initiating immunity and unresponsiveness. (10) Transplanting tissues. (11) Studying items closely related to immunity such as complement, reticuloendothelial system and others. (12) Molecular techniques for studying immune cells and their receptors. (13) Imaging of the immune system. (14) Methods for production or their fragments in eukaryotic and prokaryotic cells. In addition the journal will publish articles on novel methods for analysing the organization, structure and expression of genes for immunologically important molecules such as immunoglobulins, T cell receptors and accessory molecules involved in antigen recognition, processing and presentation. Submitted full length manuscripts should describe new methods of broad applicability to immunology and not simply the application of an established method to a particular substance - although papers describing such applications may be considered for publication as a short Technical Note. Review articles will also be published by the Journal of Immunological Methods. In general these manuscripts are by solicitation however anyone interested in submitting a review can contact the Reviews Editor and provide an outline of the proposed review.