Additional file 1 of Regulatory and effector T cell subsets in tumor-draining lymph nodes of patients with squamous cell carcinoma of head and neck

Additional file 1: Fig. S1. Flow cytometry analysis of unstimulated CD4+ T cell subsets in the tumor draining lymph nodes of patients with HNSCC. Lymphocytes were gated (A) CD4+ T cells were gated followed by defining these subpopulations in CD4+ T cells gate: (B) TNF-α+ cells (C) IFN-γ+ cells (D) IL-4+ cells (E) IL-17+ cells (F) IL-10+ cells (G) TGF-β+ cells subsets according to their cognate cytokine expression. Fig. S2. Flow cytometry analysis of unstimulated CD8+ T cell subsets in the tumor draining lymph nodes of patients with HNSCC. (A) Lymphocytes were gated (B) CD8+ T cells were gated followed by defining these subpopulations in CD8+ T cells gate: (C) TNF-α+ cells (D) IFN-γ+ cells (E) IL-4+ cells (E) IL-17+ cells (F) IL-10+ cells (G) TGF-β+ cells subsets according to their cognate cytokine expression. Fig. S3. Comparison of the frequencies of CD4+ and its subsets in metastatic (MLNs) and non-metastatic lymph nodes (nMLNs) of patients with tongue SCC. CD4+ T cells gated on the lymphocyte population and then the frequencies of T cell subpopulations were determined within CD4+ T cell gate. Horizontal bar is representative of the Mean ± SEM. *P value < 0.05. Fig. S4. Comparison of the frequencies of CD4+ Foxp3+ subsets in patients with tongue SCC with at least one involved lymph node (LN +) or without lymph node involvement (LN‒). CD4+ T cells gated on the lymphocyte population and then the frequencies of T cell subpopulations were determined within CD4+ T cell gate. Horizontal bar is representative of the Mean ± SEM. *P value < 0.05. Fig. S5. Comparison of the frequencies of CD8+ and its subsets in metastatic (MLNs) and non-metastatic lymph nodes (nMLNs) of patients with tongue SCC. CD8+ T cells gated on the lymphocyte population and then the frequencies of T cell subpopulations were determined within CD8+ T cell gate. Horizontal bar is representative of the Mean ± SEM. *P value < 0.05. Fig. S6. Comparison of CD8+ T cells and its subsets in patients with tongue SCC with tumor grade I and grade II + III. CD8+ T cells gated on the lymphocyte population and then the frequencies of T cell subpopulations were determined within CD8+ T cell gate. Horizontal bar is representative of the Mean ± SEM, *P value < 0.05. Fig. S7. Comparison of gMFI of TNF-α, IFN-γ, IL-4, IL-17, IL-10 and TGF-β in CD4+ lymphocytes in TDLNs of tongue SCC with or without perineural and/or lymphovascular invasion. Horizontal bar is representative of the Mean ± SEM. *P value < 0.05. Fig. S8. Comparison of CD8+ T cells and its subsets in TDLNs of tongue and laryngeal SCC with the stage IV of disease. Horizontal bar is representative of the Mean ± SEM, *P value < 0.05. Fig. S9. Comparison of CD4+Foxp3+T cells subsets in TDLNs of tongue and laryngeal SCC without LN involvement. Horizontal bar is representative of the Mean ± SEM, *P value < 0.05. Fig. S10. Comparison of CD4+ T cells subsets in TDLNs of tongue and laryngeal SCC without LN involvement. Horizontal bar is representative of the Mean ± SEM, *P value < 0.05, **P value < 0.01. Fig. S11. Comparison of CD4+ T cells and its subsets in TDLNs of tongue and laryngeal SCC with tumor size ≤ 3 cm. Horizontal bar is representative of the Mean ± SEM, *P value < 0.05. Fig. S12. Comparison of CD8+ T cells and its subsets in TDLNs of tongue and laryngeal SCC with tumor size ≤ 3 cm. Horizontal bar is representative of the Mean ± SEM, **P value < 0.01. Fig. S13. Comparison of CD4+Foxp3.+T cells subsets in TDLNs of tongue and laryngeal SCC with tumor size ≤ 3 cm. Horizontal bar is representative of the Mean ± SEM, *P value < 0.0