Development and application of an end-to-end staining and analysis pipeline to identify immune cell infiltrates in oral cancer samples using a targeted multiplex immunohistochemistry antibody panel

Danielle Fails, Trevor D. McKee, Michael Spencer

The tumor microenvironment (TME) plays a critical role in determining how a tumor may respond to therapy, particularly for cancer immunotherapy approaches. Improvements in the number of biomarkers that can be simultaneously screened have led to the ability to probe for numerous cell types expressing multiple markers within the TME. The ability to probe the tumor microenvironment (TME) at the single cell level is important for understanding of interactions between tumors and immune cells, particularly whether immune cells have infiltrated into tumor nests, and with regards to the presence or absence of immune checkpoint markers. The ability to interrogate these questions relies on the ability to assess multiple biomarkers simultaneously for cell phenotyping in their spatial context. Here we demonstrate rapid design and optimization of a panel of antibodies for multiplexed immunohistochemical staining of a series of oral cancer tumor samples.