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Sequential Chromogenic IHC: Spatial Analysis of Lymph Nodes Identifies Contact Interactions between Plasmacytoid
Natalie Claudio1,2, My-Tien Nguyen3, Adrian Wanner4
1Department of Otolaryngology - Head and Neck Surgery, Oregon Health & Science University, Portland, Oregon.
Insights
This study offers a guide for creating custom biomarker panels for sequential chromogenic immunohistochemistry (scIHC) to analyze immune cells in cancer. The method identified new interactions between plasmablasts and plasmacytoid dendritic cells in head and neck cancer patients.
Area of Science:
- Immunology
- Oncology
- Biotechnology
Background:
- Spatial organization of immune cells in lymphoid structures is crucial for cancer immunotherapy success and patient survival.
- Sequential chromogenic immunohistochemistry (scIHC) is limited by a lack of standardized guides for biomarker panel development and user-friendly analysis pipelines.
Purpose of the Study:
- To provide a comprehensive guide for developing novel biomarker panels for scIHC.
- To present a user-friendly analysis pipeline for scIHC data.
- To investigate cell-cell interactions in head and neck cancer using scIHC.
Main Methods:
- Development of a standardized guide for scIHC biomarker panel creation.
- Detailed instructions to prevent and detect cross-reactivity and antibody carryover.
- Creation of an analysis pipeline with non-rigid tissue deformation correction, automated cell segmentation (Cellpose-inspired), and computational masking.
- Application of the panel and pipeline to regional lymph nodes from head and neck cancer patients.
Main Results:
- Successful development of customized biomarker panels for scIHC.
- Identification of novel in vivo contact interactions between plasmablasts and plasmacytoid dendritic cells in head and neck cancer lymph nodes.
- Demonstration of a streamlined approach for scIHC data analysis.
Conclusions:
- The provided guide and analysis pipeline facilitate the development of scIHC biomarker panels.
- The findings highlight the significance of plasmablast and plasmacytoid dendritic cell interactions in cancer immunity.
- This work improves the understanding of immune responses in cancer through advanced scIHC techniques.
Abstract:
Recent clinical observations have emphasized the critical role that the spatial organization of immune cells in lymphoid structures plays in the success of cancer immunotherapy and patient survival. However, implementing sequential chromogenic IHC (scIHC) to analyze multiple biomarkers on a single tissue section has been limited because of a lack of a standardized, rigorous guide to the development of customized biomarker panels and a need for user-friendly analysis pipelines that can extract meaningful data. In this context, we provide a comprehensive guide for the development of novel biomarker panels for scIHC, using practical examples and illustrations to highlight the most common complications that can arise during the setup of a new biomarker panel, and provide detailed instructions on how to prevent and detect cross-reactivity between secondary reagents and carryover between detection antibodies. We also developed a novel analysis pipeline based on non-rigid tissue deformation correction, Cellpose-inspired automated cell segmentation, and computational network masking of low-quality data. We applied this biomarker panel and pipeline to study regional lymph nodes from patients with head and neck cancer, identifying novel contact interactions between plasmablasts and plasmacytoid dendritic cells in vivo. Given that Toll-like receptors, which are highly expressed in plasmacytoid dendritic cells, play a key role in vaccine efficacy, the significance of this cell-cell interaction decisively warrants further studies. In summary, this work provides a streamlined approach to the development of customized biomarker panels for scIHC that will ultimately improve our understanding of immune responses in cancer.
Significance:
We present a comprehensive guide for developing customized biomarker panels to investigate cell-cell interactions in the context of immune responses in cancer. This approach revealed novel contact interactions between plasmablasts and plasmacytoid dendritic cells in lymph nodes from patients with head and neck cancer.
