Tumor-infiltrating lymphocyte subsets and tertiary lymphoid structures in pulmonary metastases from colorectal cancer

Abstract

The presence of tumor-infiltrating lymphocytes (TILs) and tertiary lymphoid structures (TLSs) reflects an active inflammatory tumor microenvironment. High density of TILs as well as presence of TLS is associated with improved survival in various solid cancer types. We aimed to describe the density and distribution of TILs and TLS in pulmonary metastases (PMs) from primary colorectal cancer (CRC) and its correlation with clinicopathological variables. Fifty-seven CRC pulmonary metastasectomy specimen (PM) and 31 matched primary CRC specimen were included. Cluster of differentiation (CD)3+, CD8+, CD45RO+ and FoxP3+ TILs were evaluated by immunohistochemistry and density was scored semiquantitatively. TLS were evaluated based on morphological criteria. Survival time was defined from pulmonary metastasectomy to death or last follow up. A marked infiltration with CD3+, CD8+, CD45RO+ and FoxP3+ TILs was evident in CRC PM and matched primary CRC. Further assessment of the immune infiltrate in PM showed that a high density of FOXP3+ TILs at the invasive margin [HR 2.40 (1.11-6.96); P = 0.031] and low density of CD8+ cells in TLS [HR 0.30 (0.14-0.79); P = 0.016] were associated with a worse prognosis in univariate analysis. Moreover, a low CD8/FoxP3-ratio of TILs at the invasive margin (P = 0.042) and in TLS (P = 0.027) conferred an impaired prognosis after pulmonary metastasectomy. Our findings suggest that CRC PM harbor an immune active microenvironment. The balance of CD8+ and FoxP3+ T-cells at the tumor border and in TLS provides prognostic information in patients with CRC PM.

Keywords: Colorectal cancer; Pulmonary metastasis; TILs; Tertiary lymphoid structures; Tumor-infiltrating lymphocytes.

Fig. 1

Representative images of high densities of a CD3+, b CD8+, c CD45RO+ and d FoxP3+ TILs (DAB; brown) in pulmonary metastases. e CD3+ tertiary lymphoid structure at the invasive margin between tumor cells and lung parenchyma (magnification ×200/400). (Color figure online)

Fig. 2

Kaplan–Meier estimates regarding recurrence-free survival and overall survival of pulmonary metastases dependent on the density of CD8+ and FoxP3+ TILs. Additionally the outcome for the CD8/FoxP3-ratio was calculated

Fig. 3

Kaplan–Meier estimates regarding recurrence-free survival and overall survival of pulmonary metastases dependent on the density of CD8+ and FoxP3+ cells in TLS. Additionally the outcome for the CD8/FoxP3-ratio was calculated