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Updated: Nov 12, 2025
Multiplexed Fluorescent Immunohistochemical Staining, Imaging, and Analysis in Histological Samples of Lymphoma
Published on: January 9, 2019
Multiplexed histology analyses for the phenotypic and spatial characterization of human innate lymphoid cells
Anna Pascual-Reguant1,2, Ralf Köhler2, Ronja Mothes1,2
1Charité - Universitätsmedizin Berlin, Department of Rheumatology and Clinical Immunology, 10117, Berlin, Germany.
Insights
Innate lymphoid cells (ILCs) are key immune regulators. This study developed a new method to identify ILCs in tissues, revealing their location and interactions with other cells like plasma cells.
Area of Science:
- Immunology
- Cell Biology
- Computational Biology
Background:
- Innate lymphoid cells (ILCs) are crucial immune regulators, but their tissue localization and microenvironmental interactions remain poorly understood.
- Existing methods lack the precision to fully characterize ILCs within their native tissue context.
Purpose of the Study:
- To develop and validate a novel multiplexed immunofluorescence and computational analysis pipeline for in situ identification and characterization of CD127+ ILCs.
- To elucidate the microenvironmental niches and cellular interactions of ILCs within human tonsils.
Main Methods:
- Multiplexed immunofluorescence staining of human tonsil tissue.
- Development of a customized, open-source computational pipeline for image analysis.
- In situ identification and characterization of CD127+ ILCs and their surrounding stromal and immune cells.
Main Results:
- Successfully identified and characterized CD127+ ILCs in situ, providing detailed spatial and phenotypic information.
- Identified the transcription factor IRF4 as a specific marker for tonsillar ILC3s.
- Discovered conserved stromal landmarks associated with ILC localization and observed ILCs co-localizing with plasma cells in tonsillar niches.
Conclusions:
- The developed platform enables high-resolution, multiparametric histological analysis of ILCs in situ.
- This approach enhances our understanding of ILC biology, tissue localization, and interactions within the immune microenvironment.
- Findings provide a foundation for further research into ILC function in health and disease.
Abstract:
Innate lymphoid cells (ILCs) emerge in the last few years as important regulators of immune responses and biological processes. Although ILCs are mainly known as tissue-resident cells, their precise localization and interactions with the microenvironment are still unclear. Here we combine a multiplexed immunofluorescence technique and a customized computational, open-source analysis pipeline to unambiguously identify CD127+ ILCs in situ and characterize these cells and their microenvironments. Moreover, we reveal the transcription factor IRF4 as a marker for tonsillar ILC3, and identify conserved stromal landmarks characteristic for ILC localization. We also show that CD127+ ILCs share tissue niches with plasma cells in the tonsil. Our works thus provide a platform for multiparametric histological analysis of ILCs to improve our understanding of ILC biology.
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