Functional status and spatial architecture of tumor-infiltrating CD8+ T cells are associated with lymph node metastases in non-small cell lung cancer

Abstract

Background Anti-PD-(L)1 immunotherapy has been recommended for non-small cell lung cancer (NSCLC) patients with lymph node metastases (LNM). However, the exact functional feature and spatial architecture of tumor-infiltrating CD8 + T cells remain unclear in these patients. Methods Tissue microarrays (TMAs) from 279 IA-IIIB NSCLC samples were stained by multiplex immunofluorescence (mIF) for 11 markers (CD8, CD103, PD-1, Tim3, GZMB, CD4, Foxp3, CD31, αSMA, Hif-1α, pan-CK). We evaluated the density of CD8 + T-cell functional subsets, the mean nearest neighbor distance (mNND) between CD8 + T cells and neighboring cells, and the cancer-cell proximity score (CCPS) in invasive margin (IM) as well as tumor center (TC) to investigate their relationships with LNM and prognosis. Results The densities of CD8 + T-cell functional subsets, including predysfunctional CD8 + T cells (Tpredys) and dysfunctional CD8 + T cells (Tdys), in IM predominated over those in TC (P < 0.001). Multivariate analysis identified that the densities of CD8 + Tpredys cells in TC and CD8 + Tdys cells in IM were significantly associated with LNM [OR = 0.51, 95%CI (0.29–0.88), P = 0.015; OR = 5.80, 95%CI (3.19–10.54), P < 0.001; respectively] and recurrence-free survival (RFS) [HR = 0.55, 95%CI (0.34–0.89), P = 0.014; HR = 2.49, 95%CI (1.60–4.13), P = 0.012; respectively], independent of clinicopathological factors. Additionally, shorter mNND between CD8 + T cells and their neighboring immunoregulatory cells indicated a stronger interplay network in the microenvironment of NSCLC patients with LNM and was associated with worse prognosis. Furthermore, analysis of CCPS suggested that cancer microvessels (CMVs) and cancer-associated fibroblasts (CAFs) selectively hindered CD8 + T cells from contacting with cancer cells, and were associated with the dysfunction of CD8 + T cells. Conclusion Tumor-infiltrating CD8 + T cells were in a more dysfunctional status and in a more immunosuppressive microenvironment in patients with LNM compared with those without LNM.

FOXP3: A Player of Immunogenetic Architecture in Lung Cancer

“…The shorter distance between regulatory CD4+ and intratumoral CD8+ T cells was associated with a worse prognosis. In addition, the density of regulatory CD4+ T cells in the tumor center is predominant over those in the invasive margin; however, the difference was not statistically significant [49]. To conclude, the NSCLC patients with lymph node metastases are characterized by the high density of intratumoral-infiltrating CD8+ T cells and more immunosuppressive TME-approaching CD8+ T cells [49].…”

“…The machine learning-assisted image analysis was applied to tissue microarrays from 1116 tissue sites of 279 IA-IIIB NSCLS patients, stained by multiplex immunofluorescence for CD8, CD103, PD-1, Tim3, GZMB, CD4, Foxp3, CD31, αSMA, Hif-1α and pan-CK markers [49]. The authors observed a higher density of dysfunctional CD8+ T cells in the invasive margin compared to the tumor center.…”

“…The authors observed a higher density of dysfunctional CD8+ T cells in the invasive margin compared to the tumor center. An evaluation of the mean nearest neighbor distance (mNND) revealed a stronger immunomodulatory network in NSCLC patients with lymph node metastases (LNMs) [49]. The mNND between regulatory CD4+ and intratumoral CD8+ T cells was shorter in patients with LNM compared to those without LNM.…”

Advances in lymphatic metastasis of non-small cell lung cancer

“…Tumor-infiltrating CD8 + T cells become dysfunctional and more immunosuppressive. PD-1 + CD + 8 T lymphocytes might be immune checkpoint blockade (ICB) -responsive and essential for anti-tumor immunity [ 107 – 109 ]. Moreover, host regulatory T cells (Treg) could induce conversion of naive tumor antigen-specific CD4 + T cells into anergy and peripherally induced Tregs in a genetically engineered lung adenocarcinoma mouse model.…”

Advancing Understanding of Non-Small Cell Lung Cancer with Multiplexed Antibody-Based Spatial Imaging Technologies

“…A pro-inflammatory function, divergent from that of T reg cells, was proposed in view of their TNF-α production, and negativity for CD127 also indicated they were distinct from innate lymphoid cells [72]. A CD3-CD4+CD127+ population was previously identified in autoimmune diseases such as rheumatoid arthritis and psoriasis; despite its T-cell lineage, this was activated by innate signals such as IL-7, which can downregulate CD127 expression in CD3-CD4+ cells [73,74].…”