# Influence and Mechanism of Immunophenotyping on the Efficacy of Neoadjuvant Therapy in Non-small Cell Lung Cancer

## TL;DR

This study shows that a specific immune cell pattern in lung cancer patients predicts better response to a combination of immunotherapy and chemotherapy.

## Contribution

The study identifies a novel immune infiltration subtype (inflammatory type) as an independent predictor of treatment response in NSCLC patients.

## Key findings

- Inflammatory-type NSCLC patients had a significantly higher major pathological response rate (71.4%) compared to non-inflammatory types (33.3%).
- Inflammatory type was confirmed as an independent protective factor for achieving major pathological response.
- In inflammatory-type NSCLC, MPR groups had lower effective density of cytotoxic CD8+ T cells near regulatory CD4+ T cells in epithelial regions.

## Abstract

免疫治疗反应率低，可部分归因于异质性肿瘤微环境导致的肿瘤免疫逃逸机制，本研究旨在通过组织学层面确定炎症型非小细胞肺癌（non-small cell lung cancer, NSCLC）对新辅助免疫联合化疗疗效的影响，并探究特定CD8+ T及CD4+ T细胞的数量和空间接近性在疗效预测中的价值。

回顾性纳入2021年1月至2023年6月在山东省肿瘤医院接受新辅助免疫联合化疗的43例NSCLC患者，收集术前活检标本并进行多重免疫荧光染色[CD8/程序性细胞死亡受体1（programmed cell death protein 1, PD-1）/T细胞免疫球蛋白及黏蛋白结构域蛋白 3（T-cell immunoglobulin and mucin-domain containing protein 3, TIM-3）/CD4/叉头框蛋白3（forkhead box protein 3, FoxP3）/细胞角蛋白（cytokeratin, CK）/4',6-二脒基-2-苯基吲哚（4',6-diamidino-2-phenylindole, DAPI）]。使用InForm软件行组织分割（上皮区和间质区），并量化肿瘤细胞、CD8+ T细胞及分群（细胞毒性、预耗竭和耗竭）、CD4+ T细胞及分群（常规和调节性）的密度及其空间接近性。基于CD8+ T细胞在上皮及间质区中的相对浸润程度，将NSCLC分为炎症型（上皮区及间质区均>10/1000）、排除型（上皮区<10/1000，间质区>10/1000）和荒漠型（上皮区及间质区均<10/1000）。使用Kolmogorov-Smirnov检验、Fisher's精确检验、Mann-Whitney U检验及Logistic回归确定与主要病理缓解（major pathological response, MPR）有关的因素。

炎症型、排除型及荒漠型NSCLC分别占65.1%、27.9%和7.0%。与非炎症型NSCLC患者相比，炎症型MPR率更高（71.4% vs 33.3%, P=0.016）。单因素以及多因素Logistic回归分析均证实炎症型是NSCLC患者获得MPR的独立保护因素（OR=0.20，95%CI: 0.05-0.74，P=0.020；校正后OR=0.17，95%CI: 0.03-0.80，P=0.030）。在炎症型NSCLC上皮区内分析CD8+ T与CD4+ T细胞空间距离时发现，MPR组上皮区内调节性CD4+ T细胞30 μm半径内细胞毒性CD8+ T细胞的有效密度低于non-MPR组（0.00 vs 0.33, P=0.037）。

炎症型NSCLC患者接受新辅助免疫联合化疗的疗效更佳，这可能与调节性CD4+ T细胞与细胞毒性CD8+ T细胞接近性减弱有关。

Clinicopathological characteristics of NSCLC patients receiving neoadjuvant immunotherapy combined with chemotherapy (n=43)

Relationship between tumor immune infiltration subtypes and clinical-pathological characteristics

Logistic regression analysis of related factors and treatment response

Analysis of differences in cell proportions between non-MPR and MPR groups in inflamed NSCLC

Analysis of differences in effective density between non-MPR and MPR groups in inflamed NSCLC