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Multiplexed Immunofluorescence Analysis and Quantification of Intratumoral PD-1+ Tim-3+ CD8+ T Cells
Published on: February 8, 2018
Spatial tertiary lymphoid structures imply response to anti-PD-1 plus anlotinib in advanced non-small cell lung
Jianli Ma1, Yuwei Deng2, Minghui Zhang2
1Department of Radiation Oncology, Harbin Medical University Cancer Hospital, Harbin, Heilongjiang Province, People's Republic of China.
Abstract:
Despite breakthroughs of immunotherapy synergistically combined with blockade of vascular endothelial growth factor receptor, several patients with advanced non-small cell lung cancer (NSCLC) experience non-response or followed relapse. Organized lymphoid aggregates, termed tertiary lymphoid structures (TLSs), are found to be associated with improved response to immunotherapy. Here, we explore the landscapes of TLSs in tumour tissues from a real-world retrospective study. Our investigation showed that with a median follow-up of 11.2 months, the ORR was 28.6% (18/63, 95% CI 17.9-41.3) and the median PFS was 6.1 (95% CI 5.5-6.6) months in NSCLC patients treated with PD-1 blockade combined with anlotinib. By multiplex immunofluorescence (mIF) analysis, spatially, more TLSs and high CD20+ B-cell ratio in TLSs were associated with higher ORR. High density of intratumoral CD8+ T cells showed better ORR and PFS. The numbers of CD8+ T cells with a distance within 20 μm and 20-50 μm between tumour cells were higher in responders than non-responders. But responders had significantly higher TLSs within 20 μm rather than within 20-50 μm of tumour cells than non-responders. The inflamed immunophenotyping occupied higher proportions in responders and was associated with better PFS. Besides, tumour cells in non-responders were found more temporal cell-in-cell structures than responders, which could protect inner cells from T-cell attacks. Taken together, landscape of TLSs and proximity architecture may imply superior responses to PD-1 blockade combined with anlotinib for patients with advanced non-small cell lung cancer.
Insights
Tertiary lymphoid structures (TLSs) in tumors correlate with better outcomes for advanced non-small cell lung cancer (NSCLC) patients treated with PD-1 blockade and anlotinib. Higher TLS density and specific immune cell proximity predict improved response and survival.
Area of Science:
- Oncology
- Immunology
- Cancer Research
Background:
- Advanced non-small cell lung cancer (NSCLC) patients often show limited response or relapse despite combined immunotherapy and VEGF receptor blockade.
- Tertiary lymphoid structures (TLSs), organized lymphoid aggregates in tumors, are linked to improved immunotherapy response.
- Understanding the tumor microenvironment, including TLSs, is crucial for optimizing NSCLC treatment strategies.
Purpose of the Study:
- To investigate the landscape of tertiary lymphoid structures (TLSs) in tumor tissues from advanced NSCLC patients.
- To correlate TLS characteristics and immune cell distribution with treatment response to PD-1 blockade combined with anlotinib.
- To identify predictive biomarkers for response in NSCLC patients undergoing this combination therapy.
Main Methods:
- Retrospective analysis of real-world NSCLC patient data.
- Multiplex immunofluorescence (mIF) to analyze TLS density, CD20+ B-cell ratios, and CD8+ T-cell proximity to tumor cells.
- Evaluation of tumor immunophenotyping and cell-in-cell structures in relation to treatment outcomes.
Main Results:
- An overall response rate (ORR) of 28.6% and median progression-free survival (PFS) of 6.1 months were observed.
- Higher TLS density and a greater CD20+ B-cell ratio within TLSs were associated with increased ORR.
- Increased intratumoral CD8+ T-cell density and proximity to tumor cells, along with inflamed immunophenotyping, correlated with better ORR and PFS. Responders showed significantly more TLSs within 20μm of tumor cells.
Conclusions:
- The spatial distribution and composition of TLSs, along with immune cell proximity, are significant predictors of response to PD-1 blockade plus anlotinib in advanced NSCLC.
- Tumor cell-in-cell structures may represent a mechanism of resistance by shielding inner cells from T-cell attack.
- TLS landscape and proximity architecture offer potential biomarkers for predicting treatment efficacy in NSCLC.
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