The Society for Immunotherapy of Cancer statement on best practices for multiplex immunohistochemistry (IHC) and immunofluorescence (IF) staining and validation
Open Access
- 15 May 2020
- journal article
- research article
- Published by BMJ in Journal for ImmunoTherapy of Cancer
- Vol. 8 (1), e000155
- https://doi.org/10.1136/jitc-2019-000155
Abstract
Objectives The interaction between the immune system and tumor cells is an important feature for the prognosis and treatment of cancer. Multiplex immunohistochemistry (mIHC) and multiplex immunofluorescence (mIF) analyses are emerging technologies that can be used to help quantify immune cell subsets, their functional state, and their spatial arrangement within the tumor microenvironment. Methods The Society for Immunotherapy of Cancer (SITC) convened a task force of pathologists and laboratory leaders from academic centers as well as experts from pharmaceutical and diagnostic companies to develop best practice guidelines for the optimization and validation of mIHC/mIF assays across platforms. Results Representative outputs and the advantages and disadvantages of mIHC/mIF approaches, such as multiplexed chromogenic IHC, multiplexed immunohistochemical consecutive staining on single slide, mIF (including multispectral approaches), tissue-based mass spectrometry, and digital spatial profiling are discussed. Conclusions mIHC/mIF technologies are becoming standard tools for biomarker studies and are likely to enter routine clinical practice in the near future. Careful assay optimization and validation will help ensure outputs are robust and comparable across laboratories as well as potentially across mIHC/mIF platforms. Quantitative image analysis of mIHC/mIF output and data management considerations will be addressed in a complementary manuscript from this task force.Keywords
This publication has 68 references indexed in Scilit:
- Highly multiplexed single-cell analysis of formalin-fixed, paraffin-embedded cancer tissueProceedings of the National Academy of Sciences of the United States of America, 2013
- Fiji: an open-source platform for biological-image analysisNature Methods, 2012
- TrakEM2 Software for Neural Circuit ReconstructionPLOS ONE, 2012
- Antigen Retrieval ImmunohistochemistryJournal of Histochemistry & Cytochemistry, 2011
- Antibody validationBioTechniques, 2010
- Molecular mechanisms of antigen retrieval: antigen retrieval reverses steric interference caused by formalin-induced cross-linksBiotechnic & Histochemistry, 2009
- Antibody Elution Method for Multiple Immunohistochemistry on Primary Antibodies Raised in the Same Species and of the Same SubtypeJournal of Histochemistry & Cytochemistry, 2009
- Molecular mechanisms of antigen retrieval: antigen retrieval reverses steric interference caused by formalin-induced cross-linksBiotechnic & Histochemistry, 2009
- Technical pitfalls potentially affecting diagnoses in immunohistochemistryJournal of Clinical Pathology, 2008
- Spectral imaging: Principles and applicationsCytometry Part A, 2006