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Author Spotlight: Unlocking Insights into the Immune Cell Landscape of Tumors
Published on: August 18, 2023
Rapid multiplex immunohistochemistry for characterizing tumor-immune microenvironment
Alisa Kimura1, Takahiro Tsujikawa1,2, Hiroki Morimoto1
1Department of Otolaryngology-Head and Neck Surgery, Kyoto Prefectural University of Medicine, Kyoto, Japan.
Insights
A new rapid multiplex immunohistochemistry (IHC) method uses 6 markers for faster, quantitative analysis of tumor-infiltrating immune cells. This technique aids in developing personalized cancer treatments by assessing the tumor microenvironment efficiently.
Area of Science:
- Oncology
- Immunology
- Biotechnology
Background:
- Intratumoral immune profiles impact cancer prognosis and treatment efficacy.
- Personalized medicine relies on identifying predictive biomarkers.
- Multiplex immunohistochemistry (IHC) is a standard for tissue evaluation.
Purpose of the Study:
- To develop a rapid, quantitative multiplex IHC method for evaluating intratumoral immune profiles.
- To adapt a previously developed 14-marker multiplex IHC into a faster 6-marker assay.
- To enable clinically feasible assessment of the tumor-immune microenvironment.
Main Methods:
- Reduced marker labels from 14 to 6 based on prognostic relevance.
- Optimized staining protocols (temperature, chromogen, washing time) for speed.
- Validated the 6-marker rapid multiplex IHC against conventional methods in benign tonsil and head and neck cancer tissues.
Main Results:
- The 6 selected markers correlated significantly with the 14 markers for immune classification.
- The rapid 6-marker multiplex IHC assay was completed in under 6 hours.
- Significant correlations were observed in staining intensity, immune cell densities, ratios, and spatial profiles compared to conventional IHC.
Conclusions:
- The developed 6-marker rapid multiplex IHC method provides a quantitative assessment of the tumor-immune microenvironment.
- This technique can be performed on a clinically feasible time scale.
- It supports the advancement of tissue biomarker-guided therapeutic strategies for personalized cancer treatment.
Abstract:
Intratumoral immune profiles are related to prognosis and therapeutic efficacy, and could result in personalized treatments based on biomarkers. To develop a multiplex, quantitative, and rapid tissue evaluation method based on the clinically established standard immunohistochemistry (IHC), a 6-marker rapid multiplex IHC was developed based on our previously reported 14-marker multiplex IHC by reducing the number of labels and accelerating the staining procedure. First, fewer labels were required to identify the same immunological features linked to prognosis in 14-marker multiplex IHC analyses. The six selected markers showed a significant correlation with the 14 markers in the immune classification. Next, a rapid staining protocol was developed by optimizing the reaction temperature, chromogen, and washing time, allowing the completion of 6-marker analysis in 5 h and 49 min, as opposed to the several days required for conventional multiplex IHC. Validation of benign tonsil and head and neck cancer tissues revealed a significant correlation between rapid and conventional 6-makrer multiplex IHC in terms of staining intensities, densities of T cells, macrophages, lymphoid/myeloid immune cell ratios, and spatial profiles of intratumoral immune infiltrates. This method may enable quantitative assessment of the tumor-immune microenvironment on a clinically feasible time scale, which promotes the development of tissue biomarker-guided therapeutic strategies.
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