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Updated: Aug 14, 2026
Multiplexed Immunofluorescence Analysis and Quantification of Intratumoral PD-1+ Tim-3+ CD8+ T Cells
Published on: February 8, 2018
Multiplex Immunofluorescence Captures Progressive Immune Exhaustion with Advancing Penile Squamous Cell Cancer Stage
Filip Ionescu1, Jonathan Nguyen2, Carlos Moran Segura2
1Genitourinary Oncology Department, H. Lee Moffitt Cancer Center, Tampa, FL 33612, USA.
Abstract:
Penile squamous cell carcinoma (PSCC) is a rare and deadly malignancy. Therapeutic advances have been stifled by a poor understanding of disease biology. Specifically, the immune microenvironment is an underexplored component in PSCC and the activity of immune checkpoint inhibitors observed in a subset of patients suggests immune escape may play an important role in tumorigenesis. Herein, we explored for the first time the immune microenvironment of 57 men with PSCC and how it varies with the presence of human papillomavirus (HPV) infection and across tumor stages using multiplex immunofluorescence of key immune cell markers. We observed an increase in the density of immune effector cells in node-negative tumors and a progressive rise in inhibitory immune players such as type 2 macrophages and upregulation of the PD-L1 checkpoint in men with N1 and N2-3 disease. There were no differences in immune cell densities with HPV status.
Insights
This study reveals changes in the penile squamous cell carcinoma (PSCC) immune microenvironment. Immune cells and checkpoints like PD-L1 increase with advanced tumor stage, but not with human papillomavirus (HPV) status.
Area of Science:
- Oncology
- Immunology
- Pathology
Background:
- Penile squamous cell carcinoma (PSCC) is a rare but aggressive cancer with limited treatment options.
- Understanding the tumor immune microenvironment is crucial for developing novel therapeutic strategies, particularly immune checkpoint inhibitors.
- Previous research has not fully explored the immune landscape in PSCC or its correlation with human papillomavirus (HPV) status and disease progression.
Purpose of the Study:
- To investigate the immune cell composition and immune checkpoint expression within the PSCC microenvironment.
- To determine how the immune microenvironment differs based on HPV infection status and tumor stage (N0 vs. N1-N2-3).
Main Methods:
- Multiplex immunofluorescence was employed to analyze key immune cell markers in tumor samples from 57 men with PSCC.
- Immune cell densities were quantified and compared across different tumor stages and HPV infection statuses.
Main Results:
- An increased density of immune effector cells was observed in node-negative (N0) tumors.
- A progressive increase in inhibitory immune players, including type 2 macrophages and programmed death-ligand 1 (PD-L1) expression, was noted in advanced stages (N1 and N2-3 disease).
- No significant differences in immune cell densities were found in relation to HPV status.
Conclusions:
- The immune microenvironment in PSCC exhibits distinct characteristics that vary with tumor stage, suggesting a role for immune escape in disease progression.
- Upregulation of inhibitory immune cells and PD-L1 in advanced stages may represent therapeutic targets for penile cancer.
- HPV status does not appear to influence the immune cell composition in the studied cohort.
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