Characterization of CD8+ and FOXP3+ T-Cell Ratios in Tumor and Stromal Compartments of Penile Squamous Cell Carcinoma: An Analysis Across Histologic Subtypes and Grades
Restricted accessResearch articleFirst published online February, 2026
Characterization of CD8+ and FOXP3+ T-Cell Ratios in Tumor and Stromal Compartments of Penile Squamous Cell Carcinoma: An Analysis Across Histologic Subtypes and Grades
The balance between CD8+ cytotoxic T cells and FOXP3+ regulatory T cells within the tumor microenvironment is crucial in cancer progression. This study aimed to characterize CD8+/FOXP3+ T-cell ratios in tumor and stromal compartments of penile squamous cell carcinoma (SCC) across different histologic subtypes and grades.
Methods
This retrospective study analyzed 1 tissue microarray block (144 spots from 35 patients). CD8 and FOXP3 immunohistochemistry was performed. Ratios of CD8+/FOXP3+ T cells per high-power field were categorized as CD8 > FOXP3 or CD8 ≤ FOXP3 in tumor and stromal compartments. Associations with histologic subtype (including WHO classification) and grade were examined.
Results
Seventy-nine spots (55%), representing 23 unique patients, were evaluable. Median intratumoral CD8+ and FOXP3+ counts were 5 and 4 per high-power field, respectively. Stromal median counts were 22 (CD8+) and 38 (FOXP3+) per high-power field. No statistically significant associations were found between CD8+/FOXP3+ ratios and histologic grade in either tumor (P = .529) or stromal compartments (P = .660). Similarly, no significant associations were observed with histologic subtype, though a non-significant trend (P = .09) was noted between stromal ratio and subtype.
Conclusions
This study characterized CD8+ and FOXP3+ T-cell infiltrates and their ratios in penile SCC, revealing distinct patterns between tumor and stromal compartments. In this cohort, significant associations with histologic grade or subtype were not observed. These findings underscore the need for larger, prospective studies incorporating HPV status and clinical outcomes to elucidate the role of these immune ratios.
BrouwerORAlbersenMParnhamA, et al.European Association of Urology-American Society of Clinical Oncology collaborative guideline on penile cancer: 2023 update. Eur Urol. 2023;83(6):548-560.
2.
ChauxANettoGJRodríguezIM, et al.Epidemiologic profile, sexual history, pathologic features, and human papillomavirus status of 103 patients with penile carcinoma. World J Urol. 2013;31(4):861-867.
3.
FicarraVAkdumanBBouchotO, et al.Prognostic factors in penile cancer. Urology. 2010;76(2):S66-S73.
4.
HanahanDWeinbergRA. Hallmarks of cancer: the next generation. Cell. 2011;144(5):646-674.
5.
VriesHM deOttenhofSRHorenblasS, et al.Defining the tumor microenvironment of penile cancer by means of the cancer immunogram. Eur Urol Focus. 2019;5(5):718-721.
6.
FridmanWHPagèsFSautès-FridmanC, et al.The immune contexture in human tumours: impact on clinical outcome. Nat Rev Cancer. 2012;12(4):298-306.
7.
KimHKwonHJHanYB, et al.Increased CD3+ T cells with a low FOXP3+/CD8+ T cell ratio can predict anti-Pd-1 therapeutic response in non-small cell lung cancer patients. Mod Pathol. 2019;32(3):367-375.
8.
RuiterEJ dOoftMLDevrieseLA, et al.The prognostic role of tumor infiltrating T-lymphocytes in squamous cell carcinoma of the head and neck: a systematic review and meta-analysis. Oncoimmunology. 2017;6(11):e1356148.
9.
CocksMTaheriDBallMW, et al.Immune-checkpoint status in penile squamous cell carcinoma: a North American cohort. Hum Pathol. 2017;59:55-61.
10.
MontellaMSabettaRRonchiA, et al.Immunotherapy in penile squamous cell carcinoma: present or future? Multitarget analysis of programmed cell death ligand 1 expression and microsatellite instability. Front Med. 2022;9:874213. doi:https://doi.org/10.3389/fmed.2022.874213
11.
UdagerAMLiuT-YSkalaSL, et al.Frequent PD-L1 expression in primary and metastatic penile squamous cell carcinoma: potential opportunities for immunotherapeutic approaches. Ann Oncol. 2016;27(9):1706-1712.
12.
AldreesR. The peritumoral CD8+/FOXP3+ cell ratio has prognostic value in triple-negative breast cancer. Appl Immunohistochem Mol Morphol. 2023;31(9):621. doi:https://doi.org/10.1097/pai.0000000000001147
MochHAminMBBerneyDM, et al.The 2022 World Health Organization classification of tumours of the urinary system and male genital organs—part A: renal, penile, and testicular tumours. Eur Urol. 2022;82(5):458-468.
15.
ChauxA. Clinicopathologic and outcome features of superficial high-grade and deep low-grade squamous cell carcinomas of the penis. SpringerPlus. 2015;4(1):248.
16.
LadoireSArnould´LMignotG, et al.Presence of Foxp3 expression in tumor cells predicts better survival in HER2-overexpressing breast cancer patients treated with neoadjuvant chemotherapy. Breast Cancer Res Treat. 2010;125(1):65-72.
17.
LiuSFoulkesWDLeungS, et al.Prognostic significance of FOXP3 + tumor-infiltrating lymphocytes in breast cancer depends on estrogen receptor and human epidermal growth factor receptor-2 expression status and concurrent cytotoxic T-cell infiltration. Breast Cancer Res. 2014;16(5):432.
18.
YingLYanFMengQ, et al.PD-L1 expression is a prognostic factor in subgroups of gastric cancer patients stratified according to their levels of CD8 and FOXP3 immune markers. Oncoimmunology. 2018;7(6):e1433520. doi:https://doi.org/10.1080/2162402x.2018.1433520
19.
GoodenMJMBockGH deLeffersN, et al.The prognostic influence of tumour-infiltrating lymphocytes in cancer: a systematic review with meta-analysis. Br J Cancer. 2011;105(1):93-103.
20.
ShangBLiuYJiangS, et al.Prognostic value of tumor-infiltrating FoxP3+ regulatory T cells in cancers: a systematic review and meta-analysis. Sci Rep. 2015;5(1):15179. doi:https://doi.org/10.1038/srep15179
21.
LiXGruossoTZuoD, et al.Infiltration of CD8+ T cells into tumor cell clusters in triple-negative breast cancer. Proc Natl Acad Sci. 2019;116(9):3678-3687.
22.
SherryLAndersonMCowanM, et al.Abstract 2937: utilizing multiplex chromogenic IHC and digital image analysis to evaluate immune cell content and spatial distribution within NSCLC tumor tissue. Cancer Res. 2017;77(13_Supplement):2937-2937.
23.
Lequerica-FernándezPSuárez-CantoJRodriguez-SantamartaT, et al.Prognostic relevance of CD4+, CD8 + and FOXP3+ TILs in oral squamous cell carcinoma and correlations with PD-L1 and cancer stem cell markers. Biomedicines. 2021;9(6):653.
24.
WangJTianSSunJ, et al.The presence of tumour-infiltrating lymphocytes (TILs) and the ratios between different subsets serve as prognostic factors in advanced hypopharyngeal squamous cell carcinoma. BMC Cancer. 2020;20:731. doi:https://doi.org/10.1186/s12885-020-07234-0
25.
LiangYLüWZhangX, et al.Tumor-infiltrating CD8 + and FOXP3 + lymphocytes before and after neoadjuvant chemotherapy in cervical cancer. Diagn Pathol. 2018;13(1):93.
26.
ZhangBWuCZhangZ, et al. CXCL12 is associated with FoxP3+ tumor-infiltrating lymphocytes and affects the survival of patients with oral squamous cell carcinoma. Oncol Lett. 2019;18:1099-1106. doi:https://doi.org/10.3892/ol.2019.10415
27.
LohneisPBoralŞKaufmannAM, et al.Human papilloma virus status of penile squamous cell carcinoma is associated with differences in tumour-infiltrating T lymphocytes. Virchows Arch. 2014;466(3):323-331.
28.
NäsmanARomanitanMNordforsC, et al.Tumor infiltrating CD8 + and Foxp3+ lymphocytes correlate to clinical outcome and human papillomavirus (HPV) status in tonsillar cancer. PLoS ONE. 2012;7(6):e38711.
29.
WansomDLightEThomasD, et al.Infiltrating lymphocytes and human papillomavirus-16–associated oropharyngeal cancer. Laryngoscope.2011;122(1):121-127.
30.
MüllerTDemesMLehnA, et al.The peri- and intratumoral immune cell infiltrate and PD-L1 status in invasive squamous cell carcinomas of the penis. Clin Transl Oncol. 2021;24(2):331-341.
31.
OnoTAzumaKKawaharaA, et al.Predictive value of CD8/FOXP3 ratio combined with PD-L1 expression for radiosensitivity in patients with squamous cell carcinoma of the larynx receiving definitive radiation therapy. Head Neck. 2020;42(12):3518-3530.
32.
ChakrabortySBhattacharjeePPandaAK, et al.Providence of the CD25 + KIR + CD127−FOXP3−CD8+ T-cell subset determines the dynamics of tumor immune surveillance. Immunol Cell Biol. 2018;96(10):1035-1048. doi:https://doi.org/10.1111/imcb.12166
33.
YangYYangXWangY, et al.Combined consideration of tumor-associated immune cell density and immune checkpoint expression in the peritumoral microenvironment for prognostic stratification of non-small-cell lung cancer patients. Front Immunol. 2022;13:811007. doi:https://doi.org/10.3389/fimmu.2022.811007
