Immune Subtypes and Survival in Patients with Primary Glioma

Abstract Background The tumor microenvironment (TME) is a critical determinant of glioma response to therapies, including immunotherapy. We have classified the immune TME of gliomas by multiplex immunofluorescence (mIF). Methods Tissue taken at initial resection from 354 patients with newly−diagnosed glioma grades 2-4 were analyzed using three mIF panels of markers for T, B, and myeloid cells. Tumor cores were characterized by the relative abundances of: (i) 15 primary immune phenotypes, (ii) 96 secondary immune phenotypes, and, (iii) 29 biologically meaningful multi-marker immune phenotypes. Results Unsupervised cluster analysis of WHO grade 4 diffuse gliomas revealed four subtypes α, β, γ, and δ that were internally reproducible. Immune subtype α was characterized by high abundance of antigen-presenting cells (APCs) and low levels of MHC II− monocytes. Subtype β was high in regulatory T cells and myeloid cells, but low in lymphocytes with effector functions. Subtype γ displayed high abundance of immune cell phenotypes, particularly lymphocytes with effector or helper functions. Subtype δ was low in immune phenotypes, with poorer outcomes. Grade 3 tumors could also be classified into α, β, γ, and δ subtypes, indicating generalizability of these immune TME subtypes across high grade gliomas. Additionally, grade 2 tumors could be classified into subtypes ε, ρ and τ identified in grade 3 tumors. Conclusions We have identified internally reproducible criteria for classifying gliomas according to the immune microenvironment, findings that could aid our understanding of the natural progression of low- and high-grade gliomas and inform the rational application of immune-oncologic therapeutic interventions.