Spatial Distribution and Prognostic Implications of Tumor-Infiltrating FoxP3- CD4+ T Cells in Biliary Tract Cancer

Abstract

The clinical implications of tumor-infiltrating T cell subsets and their spatial distribution in biliary tract cancer (BTC) patients treated with gemcitabine plus cisplatin were investigated. Materials and Methods A total of 52 BTC patients treated with palliative gemcitabine plus cisplatin were included. Multiplexed immunohistochemistry was performed on tumor tissues, and immune infiltrates were separately analyzed for the stroma, tumor margin, and tumor core. Results The density of CD8 + T cells, FoxP3-CD4 + helper T cells, and FoxP3 + CD4 + regulatory T cells was significantly higher in the tumor margin than in the stroma and tumor core. The density of LAG3-or TIM3-expressing CD8 + T cell and FoxP3-CD4 + helper T cell infiltrates was also higher in the tumor margin. In extrahepatic cholangiocarcinoma, there was a higher density of T cell subsets in the tumor core and regulatory T cells in all regions. A high density of FoxP3-CD4 + helper T cells in the tumor margin showed a trend toward better progression-free survival (PFS) (p = 0.092) and significantly better overall survival (OS) (p=0.012). In multivariate analyses, a high density of FoxP3-CD4 + helper T cells in the tumor margin was independently associated with favorable PFS and OS. Conclusion The tumor margin is the major site for the active infiltration of T cell subsets with higher levels of LAG3 and TIM3 expression in BTC. The density of tumor margin-infiltrating FoxP3-CD4 + helper T cells may be associated with clinical outcomes in BTC patients treated with gemcitabine plus cisplatin.

The role of tumor-infiltrating lymphocytes in cholangiocarcinoma

“…In 11 of the 33 studies, the distribution of TILs between peritumoral and intratumoral areas in CCA was studied [ 17 , 18 , 20 , 23 , 26 , 27 , 29 – 33 ] (Table 2 ). For iCCA, 5 studies show that CD8+, CD4+ and CD3+ T cells mainly distributed around the cancer itself [ 17 , 18 , 26 , 29 , 30 ].…”

“…For CCA (including both iCCA and eCCA), three studies revealed that CD8+, CD4+ and CD3+ T cells mainly infiltrated outside of the tumor [ 26 , 31 , 32 ]. Foxp3+ T cells were also found located in peritumoral areas mostly [ 20 ]. However, another study by Zhou et al did not observe any significant difference for the location of Foxp3+ T cells [ 31 ].…”

“…Among them, 5 studies investigated the effect of CD8+ T cells with respect to spatial distribution (intratumoral (IT), peritumoral (PT) or tumor margin (TM)) on the prognosis of iCCA and all revealed that a higher number of CD8+ T cells in the tumor margin is associated with prolonged OS. Similarly, a high density of CD4+ T cells in the tumor margin was independently associated with favourable OS or DFS [ 18 , 20 ]. Four groups investigated the relationship between Foxp3+T cells and prognosis.…”

“… [ 26 ] Tian L 2020 China iCCA (322) IT CD8+ mIHC Median (cell count, x400 HPF) 27 (median) OS / TTR Patients with a high density of CD8+ T cells displayed longer OS ( p =0.006) and more favorable TTR ( p =0.003) [ 29 ] Wu H 2021 China iCCA (50) IT / TM CD8+ IHC Median (cell count, x200 HPF) 26.5 (median) OS / RPF A higher number of CD8+ cells in the tumor margin is associated with prolonged OS and RFS ( p =0.048, p =0.033). [ 20 ] Kim H 2021 Korea CCA (52) IT / TM FoxP3-CD4+ mIHC Median (cell count, x400 HPF) Not reported OS / RFS A high density of FoxP3-CD4+ cells in the tumor margin is independently associated with favorable DFS and OS ( p =0.005, p =0.004). Various studies reported oncological outcome with respect to tumor-infiltrating lymphocytes in cholangiocarcinoma CCA cholangiocarcinoma, DFS disease free survival, eCCA extrahepatic cholangiocarcinoma, FOXP3 forkhead box P3, GBAC gallbladder cancer, HPF high power field, iCCA intrahepatic cholangiocarcinoma, IT intratumoral, PT peritumoral, Ref reference, RFS relapse-free survival, RRS recurrence rate, TIL tumor-infiltrating lymphocytes, TM tumor margin …”

Dynamic increase of M2 macrophages is associated with disease progression of colorectal cancers following cetuximab-based treatment

“…Optimized fluorescent multiplexed immunohistochemistry was performed using tyramide signal amplification in the Leica Bond Rx Automated Stainer (Leica Biosystems, Newcastle, UK) as previously described 42 . Cells were stained with antibodies against CD68 (M0876, DAKO, Glostrup, Denmark), CD206 (NBP1-90020, Novus Biological, Littleton, CO, USA), PD-L1 (13684S, Abcam, Cambridge, UK) and cytokeratin (NBP2-29429, Novus, Littleton, CO, USA), and the fluorescence signals were captured with the following fluorophores: Opal 570, Opal 620, Opal 690, and Opal 780.…”