Age and sex have no impact on expression levels of markers of immune cell infiltration and immune checkpoint pathways in patients with muscle-invasive urothelial carcinoma of the bladder treated with radical cystectomy

  • Bradley C. Holland
  • Akshay Sood
  • Kristin Delfino
  • Danuta I. Dynda
  • Sophia Ran
  • Natalie Freed
  • Shaheen AlaneeEmail author
Original Article



Advanced age and female sex have been associated with worse outcomes in patients undergoing radical cystectomy for muscle-invasive bladder cancer. A reduced immune response has been implicated as a mechanism. The objective of our study was to analyze the expression patterns of various cellular proteins active in bladder cancer immune pathways, and assess the correlation between age, sex, and the expression of these immune markers.


We obtained surgical tissue samples from equally distributed male/female patients with/without lymph node metastasis who had undergone radical cystectomy for urothelial carcinoma (UC) of the bladder (n = 50). Immunohistochemistry (IHC) for CD3 (cluster of differentiation), CD4, CD8, CD56, LAG-3 (lymphocyte-activation gene), TIM-3 (T-cell immunoglobulin and mucin-domain), PD-1 (programmed death) and PD-L1 molecules was performed and scored by a single pathologist (high versus low). Spearman’s correlation and Chi square tests investigated the association between age, sex, and IHC results.


Mean age at surgery was 67 years (range 50–78 years); all patients were Caucasians. The following percent of patients scored high for a stain: 18% CD3, 10% CD4, 0% CD8, 0% CD56, 20% LAG-3, 4% TIM-3, 0% PD-1 and 0% PD-L1. There was no association between patients’ age, sex, and the expression of any of the immune markers (p > 0.05 for all).


The association between advanced age, female sex, and worse outcomes in bladder cancer may be independent of the immune pathways active in the disease that we examined in this study.


Bladder Cancer Immunohistochemistry Age Sex 



Cluster of differentiation




Lymphocyte-activation gene 3


Programmed death receptor 1


Programmed death receptor ligand 1


Surveillance, Epidemiology and End Results program


T-cell immunoglobulin and mucin-domain 3


Tumor, node, metastasis


Urothelial carcinoma of the bladder


Author contributions

BCH: data collection, manuscript writing. AS: manuscript writing. KD: statistical analysis, manuscript writing. DID: regulatory assistance. SR: immune staining, manuscript writing. NF: immune staining, manuscript writing. SA: study concept, manuscript writing, supervision.


Southern Illinois University School of Medicine research fund.

Compliance with ethical standards

Conflict of interest

The authors declare that they have no conflict of interest.

Ethical approval

This study was approved by the institutional review board of Southern Illinois University School of Medicine (5/13/2015). No human subjects or animals were involved in the study as it was done on stored tissue.

Informed consent

No informed consent was required.


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Copyright information

© Springer-Verlag GmbH Germany, part of Springer Nature 2019

Authors and Affiliations

  1. 1.Department of Surgery, Division of UrologySouthern Illinois University School of MedicineSpringfieldUSA
  2. 2.VCORE-Center for Outcomes Research, Analytics, and EvaluationVattikuti Urology Institute, Henry Ford HospitalDetroitUSA
  3. 3.Department of Medical Microbiology, Immunology and Cell BiologySouthern Illinois University School of MedicineSpringfieldUSA
  4. 4.Pathology Associates of Central IllinoisSpringfieldUSA

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