ArticlePloS one2021
Developing an enhanced 7-color multiplex IHC protocol to dissect immune infiltration in human cancers.
Article in PloS one, 2021. The graph could read no effect estimate from its abstract, so it casts no vote on the map. Cited by 27 papers, 1 of them a synthesis that pooled it.
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Who cites it
27 citing papers in PubMed, 1 synthesis or guideline pooled it, 37 citations in OpenAlex.
- Research progress and perspectives on the application of tyramide signal amplification-based multiplex immunohistochemistry/immunofluorescence: a bibliometrics analysis.Frontiers in oncology · 2024Pooled it
- A tailored in vivo CRISPR screen identifies BAP1 as a potent tumor suppressor of sarcoma.JCI insight · 2026Article
- A Multi-omics Investigation Identifies TACC3 as a Driver of Immunosuppression in Intrahepatic Cholangiocarcinoma via Activation of the STAT3-PD-L1 Axis.Journal of clinical and translational hepatology · 2026Article
- Heat-induced antigen retrieval by using Tris-EDTA solution destroys nuclear structure in certain tissues.Biochemistry and biophysics reports · 2026Article
- iCLAP: an innovative method for integrable co-detection of low-abundance antigens with high-plex immunostaining.Nature communications · 2026Article
- Ovarian tumor cells gain competitive advantage by actively reducing the cellular fitness of microenvironment cells.Nature biotechnology · 2025Article
- Major Human Leukocyte Antigen Expression in Respiratory Papillomatosis Clinical Specimens.The Laryngoscope · 2025Article
- The responses of HNSCC patients to immunotherapy are shown by two novel co-expression patterns.NPJ precision oncology · 2025Article
- Centrosome protein TAX1BP2 mediates STING-dependent immune response and potentiates anti-PD-1 efficacy in hepatocellular carcinoma.Molecular therapy : the journal of the American Society of Gene Therapy · 2025Article
- Cutaneous melanoma.Nature reviews. Disease primers · 2025Review
- Expression of T cell-related proteins in breast ductal carcinomaHistology and histopathology · 2025Article
- Exploring the characteristics of immortalized human ovarian surface epithelial cell lines.Heliyon · 2025Article
- Nose-to-brain translocation of inhaled ultrafine elongated particles: facts and mysteries.Frontiers in toxicology · 2025Article
- Increased Multiplexity in Optical Tissue Clearing-Based Three-Dimensional Immunofluorescence Microscopy of the Tumor Microenvironment by Light-Emitting Diode Photobleaching.Laboratory investigation; a journal of technical methods and pathology · 2024Article
- Exploring the Complex and Multifaceted Interplay between Melanoma Cells and the Tumor Microenvironment.International journal of molecular sciences · 2023Review
- Abnormal generation of IL-17A represses tumor infiltration of stem-like exhausted CD8BMC medicine · 2023Article
- Spatial Profile of Tumor Microenvironment in PD-L1-Negative and PD-L1-Positive Triple-Negative Breast Cancer.International journal of molecular sciences · 2023Article
- Roles of tissue-resident immune cells in immunotherapy of non-small cell lung cancer.Frontiers in immunology · 2023Review
- Bright-Field Multiplex Immunohistochemistry Assay for Tumor Microenvironment Evaluation in Melanoma Tissues.Cancers · 2022Article
- SPCS: a spatial and pattern combined smoothing method for spatial transcriptomic expression.Briefings in bioinformatics · 2022Article
Corrections and comments
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Authors and funding
5 authors at 2 institutions in 1 country.
Funding
No grant is acknowledged in the PubMed record.
Abstract
The TSA Opal multiplex immunohistochemistry (mIHC) protocol (PerkinElmer) has been used to characterize immune infiltration in human cancers. This technique allows multiple biomarkers to be simultaneously stained in a single tissue section, which helps to elucidate the spatial relationship among individual cell types. We developed and optimized two improved mIHC protocols for a 7-color panel containing 6 biomarkers (CD3, CD8, CD163, PD-L1, FoxP3, and cytokeratin (CK)) and DAPI. The only difference between these two protocols was the staining sequence of those 6 biomarkers as the first sequence is PD-L1/CD163/CD8/CK/CD3/FoxP3/DAPI and the second sequence is FoxP3/CD163/CD8/CK/CD3/PD-L1/DAPI. By comparing PD-L1/FoxP3 staining in mIHC and singleplex PD-L1/FoxP3 staining on the adjacent slide, we demonstrated that the staining sequence does not affect the staining intensity of individual biomarkers as long as a proper antigen retrieval method was used. Our study suggests that use of an antigen retrieval buffer with higher pH value (such as Tris-EDTA pH9.0) than that of the stripping buffers (such as citrate buffer pH6.0) is helpful when using this advanced mIHC method to develop panels with multiple biomarkers. Otherwise, individual biomarkers may exhibit different intensities when the staining sequence is changed. By using this protocol, we characterized immune infiltration and PD-L1 expression in head and neck squamous cell carcinoma (HNSCC), breast cancer (BCa), and non-small cell lung cancer (NSCLC) specimens. We observed a statistically significant increase in CD3+ cell populations within the stroma of NSCLC as compared to BCa and increased PD-L1+ tumor cells in HNSCC as opposed to BCa.
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Read under generation 80e0d062 · epoch 390. Bibliography from PubMed, PubMed Central and OpenAlex; grants from NIH RePORTER; trial links from ClinicalTrials.gov; estimates, votes and beliefs from the OpenQuestion graph.