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. 2021 May 21;9(5):e001772. doi: 10.1136/jitc-2020-001772

Figure 3.

Figure 3

Multiparameter flow cytometry (MFC) scatterplots demonstrate that tumor-infiltrating lymphocytes (TILs) from pulmonary metastases (PM) express higher levels of checkpoint molecules, effector cytokines and transcription factors compared with primary bone tumors. (A) Relative proportions of CD4+Foxp3+ (regulatory T cell), CD4+Foxp3-, and CD8+TILs isolated from osteosarcoma primary bone tumors (B) and PM. (B) Expression of checkpoint molecules in TILs. Given the sequential manner in which TILs may express checkpoint molecules as they become progressively exhausted, TIM-3+ and LAG-3+ populations are gated out of PD-1+ positive population. Note: For CD4+Foxp3-PD-1+LAG-3+ cells, p=0.064. (C) Expression of the transcription factors T-bet and eomesodermin (EOMES) in PD-1+TILs. Note: For CD4+Foxp3-PD-1+T-bet+ cells, p=0.073. (D) Expression of effector cytokine IFNγ in PD-1+TILs. NS=p>0.05; *p<0.05; **p<0.01. IFNγ, interferon-γ; LAG-3, lymphocyte-activation gene 3; PD-1, programmed cell death 1; TIM-3, T-cell immunoglobulin and mucin domain-containing protein 3.