Head and neck squamous cell carcinoma (HNSCC) has both human papilloma virus (HPV) positive and negative etiologies. It is one of the growing number of tumors for which an anti-tumor immune response is implicated to play a role in the course of disease as shown by responsiveness to immune-modulation using PD-1/PD-L1 inhibitors. Profiling of the tumor microenvironment including specific infiltrating lymphocyte subsets, antigen presenting cells, expression of regulatory molecules on both tumor and immune cells, and spatial relationships between cells could lead to predictive signatures for responsiveness to both conventional and immune modulatory therapies. However, such profiling has previously been hindered by the need to identify such cells “in-situ” in tissue sections using multiple markers per cell, and limitations in available technology. The current study demonstrates Cell IDx's novel UltraPlex technology, based on a simple two step protocol using hapten labeled primary antibody cocktails followed by fluor labeled anti-hapten secondary cocktails, allowing simultaneous detection of 4 markers on the same cell. Analysis of HNSCC positive and negative tissue microarrays combining UltraPlex detection of multiple markers per cell together with serial sections stained with individual phenotyping panels demonstrates a powerful technique for 12-16 plex analysis. This permits HPV status determination, immune subset profiling, quantitative marker expression, relative cellular localization and potential for correlative signature determination associated with disease outcome and response to treatment.
Citation Format: Matt Levin, Mark Lingen, David Schwartz, Helen Snyder. Immune profiling of tumor infiltrating lymphocyte subsets, myeloid cells and HPV status of HNSCC biopsies using multiplex immunofluorescence [abstract]. In: Proceedings of the Second CRI-CIMT-EATI-AACR International Cancer Immunotherapy Conference: Translating Science into Survival; 2016 Sept 25-28; New York, NY. Philadelphia (PA): AACR; Cancer Immunol Res 2016;4(11 Suppl):Abstract nr A109.
- ©2016 American Association for Cancer Research.