Zhai Jie, Tamegnon Auriole, Jiang Mei, Pandurengan Renganayaki Krishna, Parra Edwin Roger
Department of Translational Molecular Pathology, Unit 951, The University of Texas MD Anderson Cancer Center, 2130 Holcombe Blvd, Houston, 77030, TX, USA.
Lab Anim Res. 2024 Jun 14;40(1):24. doi: 10.1186/s42826-024-00210-w.
Immune profiling has become an important tool for identifying predictive, prognostic and response biomarkers for immune checkpoint inhibitors from tumor microenvironment (TME). We aimed to build a multiplex immunofluorescence (mIF) panel to apply to formalin-fixed and paraffin-embedded tissues in mice tumors and to explore the programmed cell death protein 1/ programmed cell death 1 ligand 1 (PD-1/PD-L1) axis.
An automated eight-color mIF panel was evaluated to study the TME using seven antibodies, including cytokeratin 19, CD3e, CD8a, CD4, PD-1, PD-L1, F4-80 and DAPI, then was applied in six mice lung adenocarcinoma samples. Cell phenotypes were quantified by software to explore the co-localization and spatial distribution between immune cells within the TME. This mice panel was successfully optimized and applied to a small cohort of mice lung adenocarcinoma cases. Image analysis showed a sparse degree of immune cell expression pattern in this cohort. From the spatial analysis we found that T cells and macrophages expressing PD-L1 were close to the malignant cells and other immune cells.
Comprehensive immune profiling using mIF in translational studies improves our ability to correlate the PD-1/PD-L1 axis and spatial distribution of lymphocytes and macrophages in mouse lung cancer cells to provide new cues for immunotherapy, that can be translated to human tumors for cancer intervention.
免疫谱分析已成为从肿瘤微环境（TME）中识别免疫检查点抑制剂的预测、预后和反应生物标志物的重要工具。我们旨在构建一个多重免疫荧光（mIF）检测板，应用于小鼠肿瘤的福尔马林固定石蜡包埋组织，并探索程序性细胞死亡蛋白1/程序性细胞死亡蛋白1配体1（PD-1/PD-L1）轴。
使用七种抗体（包括细胞角蛋白19、CD3e、CD8a、CD4、PD-1、PD-L1、F4-80和DAPI）对一个自动化八色mIF检测板进行评估，以研究TME，然后将其应用于六个小鼠肺腺癌样本。通过软件对细胞表型进行量化，以探索TME内免疫细胞之间的共定位和空间分布。该小鼠检测板成功优化并应用于一小群小鼠肺腺癌病例。图像分析显示该队列中免疫细胞表达模式的稀疏程度。从空间分析中我们发现，表达PD-L1的T细胞和巨噬细胞靠近恶性细胞和其他免疫细胞。
在转化研究中使用mIF进行全面的免疫谱分析提高了我们将PD-1/PD-L1轴与小鼠肺癌细胞中淋巴细胞和巨噬细胞的空间分布相关联的能力，为免疫治疗提供了新线索，可转化至人类肿瘤用于癌症干预。
