The Society for Immunotherapy of Cancer statement on best practices for multiplex immunohistochemistry (IHC) and immunofluorescence (IF) staining and validation
- J Immunother Cancer. 2020 May;8(1):e000155. doi: 10.1136/jitc-2019-000155.
- 1. Department of Dermatology, Johns Hopkins School of Medicine, Bloomberg~Kimmel Institute for Cancer Immunotherapy, Baltimore, Maryland, USA [email protected].
- 2. Tisch Cancer Institute, Icahn School of Medicine at Mount Sinai, New York, New York City, USA.
- 3. Department of Pathology, Stanford University School of Medicine, Palo Alto, California, USA.
- 4. Department of Dermatology, Johns Hopkins School of Medicine, Bloomberg~Kimmel Institute for Cancer Immunotherapy, Baltimore, Maryland, USA.
- 5. Cancer Biology Program, Stanford University School of Medicine, Palo Alto, California, USA.
- 6. Bristol-Myers Squibb, New York, New York City, USA.
- 7. Dermatopathology, Memorial Sloan-Kettering Cancer Center, New York, New York, USA.
- 8. Merck & Co Inc, Kenilworth, New Jersey, USA.
- 9. Department of Translational Molecular Pathology, University of Texas MD Anderson Cancer Center, Houston, Texas, USA.
- 10. AstraZeneca, Gaithersburg, Maryland, USA.
- 11. Department of Pathology, Yale University School of Medicine, New Haven, Connecticut, USA.
- 12. Jounce Therapeutics Inc, Cambridge, Massachusetts, USA.
- 13. Pharma Research and Early Development (pRED), Roche Innovation Center Munich, Penzberg, Germany.
- 14. Dana-Farber/Brigham and Women's Cancer Center, Boston, Massachusetts, USA.
- 15. Harvard Medical School, Boston, Massachusetts, USA.
- 16. Department of Pathology, University of Texas MD Anderson Cancer Center, Houston, Texas, USA.
- 17. Biomedical Research Centre, Royal Marsden NHS Foundation Trust, London, UK.
- 18. Providence Portland Medical Center, Portland, Oregon, USA.
- # Contributed equally.
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.