T-lymphocyte profiles differ between keratoacanthomas and invasive squamous cell carcinomas of the human skin
T-lymphocytes are involved in tumor progression and regression. Actinic keratoses (AK) are atypical proliferations of keratinocytes of the skin. Some AK progress into invasive cutaneous squamous cell carcinomas (cSCC). Keratoacanthomas (KA) are either classified as a cSCC subtype or a benign tumor with histologic resemblance to well-differentiated cSCC as it is supposed to regress spontaneously. In contrast, cSCC represent malignant tumors that may metastasize.
To compare the T-lymphocyte profiles of AK, KA and cSCC in relation to PD-L1 expression.
Tissue micro-arrays of 103 cases of AK, 43 cases of KA and 106 cases of cSCC were stained by immunohistochemistry for E-cadherin, CD3, CD4, CD8, FOXp3, and the receptor–ligand pair PD-1/PD-L1. Immunohistological scores were computationally determined to assess PD-L1 expression as well as the expression profiles of T-lymphocytes.
AK had lower numbers of CD3+ and PD-1+ cells compared to KA and lower numbers of CD3+, CD8+ and PD-1+ cells in comparison with cSCC. KA showed significantly higher numbers of CD4+ and FOXp3+ cells as well as lower numbers of CD8+ cells in comparison with invasive cSCC. cSCC expressed significantly more PD-L1 in comparison with AK and KA. Among cSCC PD-L1 expression was higher in moderately and poorly-differentiated cSCC than in well-differentiated cSCC. Increased PD-L1 expression also correlated with increased numbers of CD4+, CD8+ and FOXp3+ cells in cSCC.
Tumor-associated T-lymphocyte infiltrates showed significant differences between AK, KA and invasive cSCC. PD-L1 expression correlated with invasion of T-cell infiltrates in invasive cSCC.
KeywordsPD-1 PD-L1 FOXp3 Tumor microenvironment
Cutaneous squamous cell carcinoma
National Center of Tumor Diseases
Organ transplant recipients
Region of interest
Transforming growth factor beta
We like to thank Dr. Damir Krunic from the Imaging Core Facility of the German Cancer Research Center (Deutsches Krebsforschungszentrum, DKFZ, Germany) as well as Helmut Hübers for technical support with image analysis. This research was supported by the tissue bank of the NCT (Heidelberg, Germany) by staining the skin tumor samples.
Study conception and design were done by Corinne Bauer, Ashik Ahmed Abdul Pari, Viktor Umansky, Petra Boukamp, Cyrill Géraud and Moritz Felcht. Corinne Bauer, Ashik Ahmed Abdul Pari, Cyrill Géraud and Moritz Felcht are responsible for the integrity of acquired data. Statistical analysis was performed by Corinne Bauer, Ashik Ahmed Abdul Pari, Cyrill Géraud and Moritz Felcht. Corinne Bauer, Ashik Ahmed Abdul Pari, Viktor Umansky, Petra Boukamp, Cyrill Géraud, Moritz Felcht prepared the initial manuscript. Corinne Bauer, Ashik Ahmed Abdul Pari, Viktor Umansky, Jochen Utikal, Petra Boukamp, Hellmut G. Augustin, Sergij Goerdt, Cyrill Géraud and Moritz Felcht made substantial contributions to data analysis and interpretation of results, rewriting of the manuscript, review and approval.
This work was supported by Grants from the Deutsche Forschungsgemeinschaft (German Research Council) [Förderprojekt Nr. FE 1282/2-1 (Moritz Felcht) and GRK2099/RTG2099 “Hallmarks of Skin Cancer” (to Viktor Umansky, Jochen Utikal, Hellmut G. Augustin, Sergij Goerdt, Cyrill Géraud and Moritz Felcht)] and partially supported by the Federal Ministry for Research and Education (Bundesmininsterium für Bildung und Forschung, BMBF) (FKZ 02NUK036A to Petra Boukamp).
Compliance with ethical standards
Conflict of interest
Moritz Felcht received travel and congress participation funding by TEVA company, honoraria as an advisory board member of AbbVie Ltd. and lecture fees by Periderm GmbH and Mibe Vertrieb GmbH. All other authors declare no conflict of interest.
The study was performed with archived paraffin-embedded tissue samples. The study was approved by the ethical committee II of Heidelberg University (2014-835R-MA).
Informed consent by individual patients cannot be given, as the study only included paraffin-embedded archived tissue. With the approval of the ethical committee informed consent was not required as all patient data were anonymized.
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