Related Experiment Video
Updated: Jun 23, 2025
Author Spotlight: Enhancing Multicolor Fluorescence Localization in Lung Carcinoma Sample
Published on: November 21, 2023
Immune profiling of mouse lung adenocarcinoma paraffin tissues using multiplex immunofluorescence panel: a pilot
Jie Zhai1, Auriole Tamegnon1, Mei Jiang1
1Department of Translational Molecular Pathology, Unit 951, The University of Texas MD Anderson Cancer Center, 2130 Holcombe Blvd, Houston, 77030, TX, USA.
Insights
Multiplex immunofluorescence (mIF) profiling of mouse lung tumors reveals spatial relationships between immune cells and PD-1/PD-L1 expression. This approach enhances understanding of the tumor microenvironment for potential immunotherapy translation.
Area of Science:
- Oncology
- Immunology
- Biomarker Discovery
Background:
- Immune profiling is crucial for identifying biomarkers predictive of response to immune checkpoint inhibitors.
- The tumor microenvironment (TME) is a key area for studying these biomarkers.
- The programmed cell death protein 1/programmed cell death 1 ligand 1 (PD-1/PD-L1) axis is a significant target in immunotherapy.
Purpose of the Study:
- To develop a multiplex immunofluorescence (mIF) panel for analyzing formalin-fixed, paraffin-embedded mouse tumors.
- To investigate the PD-1/PD-L1 axis within the TME of mouse lung adenocarcinoma.
- To explore the spatial distribution and co-localization of immune cells.
Main Methods:
- An automated eight-color mIF panel was designed and validated using seven antibodies (cytokeratin 19, CD3e, CD8a, CD4, PD-1, PD-L1, F4-80) plus DAPI.
- The panel was applied to six mouse lung adenocarcinoma samples.
- Image analysis software quantified cell phenotypes, co-localization, and spatial distribution.
Main Results:
- The mIF panel was successfully optimized and applied to mouse lung adenocarcinoma samples.
- Image analysis revealed a sparse immune cell expression pattern in the studied cohort.
- Spatial analysis indicated that PD-L1 expressing T cells and macrophages were located near malignant and other immune cells.
Conclusions:
- Comprehensive immune profiling via mIF in translational research aids in correlating the PD-1/PD-L1 axis with immune cell spatial distribution in mouse lung cancer.
- This approach offers insights into immunotherapy strategies.
- Findings can potentially be translated to human tumors for cancer intervention.
Background:
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.
Results:
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.
Conclusions:
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.
More Related Videos
10:49Automated Multiplex Immunofluorescence Panel for Immuno-oncology Studies on Formalin-fixed Carcinoma Tissue Specimens
Published on: January 21, 2019
09:52DNA-barcode-based Multiplex Immunofluorescence Imaging to Analyze FFPE Specimens from Genetically Reprogrammed Murine Melanoma
Published on: June 6, 2025