{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,10,20]],"date-time":"2026-10-20T15:59:37Z","timestamp":1792511977782,"version":"4.5.0"},"reference-count":28,"publisher":"MDPI AG","issue":"3","license":[{"start":{"date-parts":[[2025,8,14]],"date-time":"2025-08-14T00:00:00Z","timestamp":1755129600000},"content-version":"vor","delay-in-days":0,"URL":"https:\/\/creativecommons.org\/licenses\/by\/4.0\/"}],"content-domain":{"domain":[],"crossmark-restriction":false},"short-container-title":["Medical Sciences"],"abstract":"<jats:p>Background: Seminoma is the most common subtype of testicular germ cell tumors in young men; however, the contribution of tumor-associated macrophages (TAMs) to disease progression remains insufficiently understood. This study aimed to quantitatively and phenotypically characterize CD68+ and CD163+ TAMs in non-metastatic seminomas (pT1N0M0 and pT2N0M0). Methods: This retrospective, multicenter, cohort, observational, analytical study was conducted from 1 January 2015 to 1 January 2025 at two branches of the National Medical Research Radiological Center of the Ministry of Health of the Russian Federation: the A. Tsyb Medical Radiological Research Center and the P. Hertsen Moscow Oncology Research Institute. Archived paraffin-embedded tumor samples from 96 patients and 21 samples of normal testicular tissue were analyzed using immunohistochemistry and digital morphometric analysis with QuPath software to assess macrophage density and spatial distribution. Results: Compared to normal testicular tissue, seminomas demonstrated more than a 10-fold increase in CD68+ TAMs and over a 100-fold increase in CD163+ TAMs. CD68+ cells predominantly localized to peripheral tumor regions, while CD163+ cells formed diffuse clusters in central tumor zones and around peripheral vessels. No statistically significant differences in CD68+ cell density were found between pT1 and pT2 stages. However, pT2 tumors showed a trend toward higher CD163+ TAMs density, suggesting increased M2 polarization with advancing tumor stage. Conclusions: These findings highlight the spatial and phenotypic heterogeneity of TAMs in seminoma and indicate a shift toward an immunosuppressive tumor microenvironment during local progression. Future studies should assess macrophage polarization and progression-free survival to evaluate their potential as prognostic biomarkers and therapeutic targets in seminoma.<\/jats:p>","DOI":"10.3390\/medsci13030129","type":"journal-article","created":{"date-parts":[[2025,8,14]],"date-time":"2025-08-14T14:51:46Z","timestamp":1755183106000},"page":"129","update-policy":"https:\/\/doi.org\/10.3390\/mdpi_crossmark_policy","source":"Crossref","is-referenced-by-count":2,"title":["Phenotypic and Spatial Characterization of Tumor-Associated Macrophages in Non-Metastatic Seminoma: Association with Local Tumor Progression"],"prefix":"10.3390","volume":"13","author":[{"ORCID":"https:\/\/orcid.org\/0000-0001-8447-2600","authenticated-orcid":false,"given":"Grigory","family":"Demyashkin","sequence":"first","affiliation":[{"name":"Department of Digital Oncomorphology, National Medical Research Centre of Radiology, 2nd Botkinsky Pass, 3, 125284 Moscow, Russia"},{"name":"Laboratory of Histology and Immunohistochemistry, Institute of Translational Medicine and Biotechnology, I.M. Sechenov First Moscow State Medical University (Sechenov University), Trubetskaya Str. 8\/2, 119048 Moscow, Russia"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0003-3763-7454","authenticated-orcid":false,"given":"Vladimir","family":"Shchekin","sequence":"additional","affiliation":[{"name":"Department of Digital Oncomorphology, National Medical Research Centre of Radiology, 2nd Botkinsky Pass, 3, 125284 Moscow, Russia"},{"name":"Research and Educational Resource Center for Immunophenotyping, Digital Spatial Profiling and Ultrastructural Analysis Innovative Technologies, Peoples\u2019 Friendship University of Russia (RUDN University), Miklukho-Maklaya Str.6, 117198 Moscow, Russia"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Dmitriy","family":"Belokopytov","sequence":"additional","affiliation":[{"name":"Department of Digital Oncomorphology, National Medical Research Centre of Radiology, 2nd Botkinsky Pass, 3, 125284 Moscow, Russia"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Tatyana","family":"Borovaya","sequence":"additional","affiliation":[{"name":"Department of Digital Oncomorphology, National Medical Research Centre of Radiology, 2nd Botkinsky Pass, 3, 125284 Moscow, Russia"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Ivan","family":"Zaborsky","sequence":"additional","affiliation":[{"name":"Department of Digital Oncomorphology, National Medical Research Centre of Radiology, 2nd Botkinsky Pass, 3, 125284 Moscow, Russia"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Kadir","family":"Safiullin","sequence":"additional","affiliation":[{"name":"Department of Digital Oncomorphology, National Medical Research Centre of Radiology, 2nd Botkinsky Pass, 3, 125284 Moscow, Russia"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Oleg","family":"Karyakin","sequence":"additional","affiliation":[{"name":"Department of Digital Oncomorphology, National Medical Research Centre of Radiology, 2nd Botkinsky Pass, 3, 125284 Moscow, Russia"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Alexey","family":"Krasheninnikov","sequence":"additional","affiliation":[{"name":"Department of Digital Oncomorphology, National Medical Research Centre of Radiology, 2nd Botkinsky Pass, 3, 125284 Moscow, Russia"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Nikolay","family":"Vorobyev","sequence":"additional","affiliation":[{"name":"Department of Digital Oncomorphology, National Medical Research Centre of Radiology, 2nd Botkinsky Pass, 3, 125284 Moscow, Russia"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Petr","family":"Shegay","sequence":"additional","affiliation":[{"name":"Department of Digital Oncomorphology, National Medical Research Centre of Radiology, 2nd Botkinsky Pass, 3, 125284 Moscow, Russia"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Andrei","family":"Kaprin","sequence":"additional","affiliation":[{"name":"Department of Digital Oncomorphology, National Medical Research Centre of Radiology, 2nd Botkinsky Pass, 3, 125284 Moscow, Russia"},{"name":"Department of Urology and Operative Nephrology, Peoples\u2019 Friendship University of Russia (RUDN University), Miklukho-Maklaya Str.6, 117198 Moscow, Russia"}],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"1968","published-online":{"date-parts":[[2025,8,14]]},"reference":[{"key":"ref_1","first-page":"10","article-title":"Cancer statistics, 2025","volume":"75","author":"Siegel","year":"2025","journal-title":"CA Cancer J. Clin."},{"key":"ref_2","doi-asserted-by":"crossref","first-page":"177","DOI":"10.5603\/FHC.a2015.0020","article-title":"Diagnostic markers for germ cell neoplasms: From placental-like alkaline phosphatase to micro-RNAs","volume":"53","author":"De","year":"2015","journal-title":"Folia Histochem. Cytobiol."},{"key":"ref_3","doi-asserted-by":"crossref","first-page":"1553","DOI":"10.1200\/JCO.20.03292","article-title":"Survival and new prognosticators in metastatic seminoma: Results from the IGCCCG-Update Consortium","volume":"39","author":"Beyer","year":"2021","journal-title":"J. Clin. Oncol."},{"key":"ref_4","doi-asserted-by":"crossref","first-page":"59","DOI":"10.1186\/s12964-020-0530-4","article-title":"Tumor microenvironment complexity and therapeutic implications","volume":"18","author":"Baghban","year":"2020","journal-title":"Cell Commun. Signal."},{"key":"ref_5","doi-asserted-by":"crossref","unstructured":"Salemme, V., Centonze, G., Cavallo, F., Defilippi, P., and Conti, L. (2021). The crosstalk between tumor cells and the immune microenvironment in breast cancer: Implications for immunotherapy. Front. Oncol., 11.","DOI":"10.3389\/fonc.2021.610303"},{"key":"ref_6","doi-asserted-by":"crossref","first-page":"1","DOI":"10.1080\/01635581.2022.2096909","article-title":"Complexity of tumor microenvironment: Therapeutic role of curcumin and its metabolites","volume":"75","author":"Prasad","year":"2023","journal-title":"Nutr. Cancer"},{"key":"ref_7","doi-asserted-by":"crossref","first-page":"373","DOI":"10.1038\/s41586-022-05254-3","article-title":"STING-induced regulatory B cells compromise NK function in cancer immunity","volume":"610","author":"Li","year":"2022","journal-title":"Nature"},{"key":"ref_8","doi-asserted-by":"crossref","first-page":"66","DOI":"10.1111\/j.1600-065X.2006.00438.x","article-title":"The testis in immune privilege","volume":"213","author":"Fijak","year":"2006","journal-title":"Immunol. Rev."},{"key":"ref_9","doi-asserted-by":"crossref","unstructured":"Savelyeva, A.V., and Medvedev, K.E. (2023). Seminoma subtypes differ in the organization and functional state of the immune microenvironment. 3 Biotech, 13.","DOI":"10.1007\/s13205-023-03530-1"},{"key":"ref_10","doi-asserted-by":"crossref","first-page":"47","DOI":"10.1016\/j.canlet.2018.05.009","article-title":"Trastuzumab upregulates PD-L1 as a potential mechanism of trastuzumab resistance","volume":"430","author":"Chaganty","year":"2018","journal-title":"Cancer Lett."},{"key":"ref_11","doi-asserted-by":"crossref","first-page":"43","DOI":"10.1111\/cas.13424","article-title":"PD-L1 expression is mainly regulated by interferon gamma in gastric cancer","volume":"109","author":"Mimura","year":"2018","journal-title":"Cancer Sci."},{"key":"ref_12","doi-asserted-by":"crossref","unstructured":"Wu, K., Lin, K., Li, X., Yuan, X., Xu, P., Ni, P., and Xu, D. (2020). Redefining Tumor-Associated Macrophage Subpopulations and Functions in the Tumor Microenvironment. Front. Immunol., 11.","DOI":"10.3389\/fimmu.2020.01731"},{"key":"ref_13","doi-asserted-by":"crossref","first-page":"1512","DOI":"10.3389\/fonc.2019.01512","article-title":"Evaluating the Polarization of Tumor-Associated Macrophages Into M1 and M2 Phenotypes in Human Cancer Tissue: Technicalities and Challenges in Routine Clinical Practice","volume":"9","author":"Jayasingam","year":"2020","journal-title":"Front. Oncol."},{"key":"ref_14","doi-asserted-by":"crossref","unstructured":"Miki, S., Suzuki Ji Takashima, M., Ishida, M., Kokubo, H., and Yoshizumi, M. (2021). S-1-Propenylcysteine promotes IL-10-induced M2c macrophage polarization through prolonged activation of IL-10R\/STAT3 signaling. Sci. Rep., 11.","DOI":"10.1038\/s41598-021-01866-3"},{"key":"ref_15","first-page":"4","article-title":"NCCN Guidelines\u00ae Insights: Testicular Cancer, Version 2.2025","volume":"23","author":"Gilligan","year":"2025","journal-title":"J. Natl. Compr. Cancer Netw."},{"key":"ref_16","doi-asserted-by":"crossref","unstructured":"Wei, Q., Deng, Y., Yang, Q., Zhan, A., and Wang, L. (2023). The markers to delineate different phenotypes of macrophages related to metabolic disorders. Front. Immunol., 14.","DOI":"10.3389\/fimmu.2023.1084636"},{"key":"ref_17","doi-asserted-by":"crossref","first-page":"387","DOI":"10.1093\/ajcp\/aqz184","article-title":"Differences in PD-L1\u2013Expressing Macrophages and Immune Microenvironment in Testicular Germ Cell Tumors","volume":"153","author":"Sadigh","year":"2020","journal-title":"Am. J. Clin. Pathol."},{"key":"ref_18","doi-asserted-by":"crossref","unstructured":"Bankhead, P., Loughrey, M., Fernandez, J., Dombrowski, Y., McArt, D., Dunne, P., McQuaid, S., Gray, R.T., Murray, L.J., and Coleman, H.G. (2017). QuPath: Open source software for digital pathology image analysis. Sci. Rep., 7.","DOI":"10.1038\/s41598-017-17204-5"},{"key":"ref_19","doi-asserted-by":"crossref","first-page":"e1440166","DOI":"10.1080\/2162402X.2018.1440166","article-title":"Neurotensin\/IL-8 pathway induces M2 polarization of TAMs and EMT in HCC","volume":"7","author":"Xiao","year":"2018","journal-title":"Oncoimmunology"},{"key":"ref_20","doi-asserted-by":"crossref","first-page":"200","DOI":"10.1097\/CMR.0000000000000352","article-title":"M1 and M2 Macrophages\u2019 Clinicopathological Significance in Cutaneous Melanoma","volume":"27","author":"Falleni","year":"2017","journal-title":"Melanoma Res."},{"key":"ref_21","doi-asserted-by":"crossref","unstructured":"Edin, S., Wikberg, M., Dahlin, A., Rutegard, J., Oberg, A., Oldenborg, P., and Palmqvist, R. (2012). The distribution of macrophages with a M1 or M2 phenotype in relation to prognosis and the molecular characteristics of colorectal cancer. PLoS ONE, 7.","DOI":"10.1371\/journal.pone.0047045"},{"key":"ref_22","doi-asserted-by":"crossref","unstructured":"Nowak, M., and Klink, M. (2020). The Role of Tumor-Associated Macrophages in the Progression and Chemoresistance of Ovarian Cancer. Cells, 9.","DOI":"10.3390\/cells9051299"},{"key":"ref_23","doi-asserted-by":"crossref","first-page":"186","DOI":"10.1186\/s12957-021-02299-y","article-title":"Prognostic significance of CD163+ tumor-associated macrophages in colorectal cancer","volume":"19","author":"Xue","year":"2021","journal-title":"World J. Surg. Oncol."},{"key":"ref_24","doi-asserted-by":"crossref","unstructured":"Shinohara, H., Kobayashi, M., Hayashi, K., Nogawa, D., Asakawa, A., Ohata, Y., Kubota, K., Takahashi, H., Yamada, M., and Tokunaga, M. (2022). CD163\u2212\/PD-L1+ TAMs as a marker of favorable outcome in TNBC. Int. J. Mol. Sci., 23.","DOI":"10.3390\/ijms232113235"},{"key":"ref_25","doi-asserted-by":"crossref","unstructured":"Jeremiasen, M., Borg, D., Hedner, C., Svensson, M., Nodin, B., Leandersson, K., Johansson, J., and Jirstr\u00f6m, K. (2020). CD68+, CD163+, and MARCO+ macrophages as prognostic biomarkers in gastroesophageal adenocarcinoma. Front. Oncol., 10.","DOI":"10.3389\/fonc.2020.534761"},{"key":"ref_26","doi-asserted-by":"crossref","unstructured":"Fei, L., Ren, X., Yu, H., and Zhan, Y. (2021). Targeting the CCL2\/CCR2 axis in cancer immunotherapy: One stone, three birds?. Front. Immunol., 12.","DOI":"10.3389\/fimmu.2021.771210"},{"key":"ref_27","doi-asserted-by":"crossref","first-page":"110297","DOI":"10.1016\/j.intimp.2023.110297","article-title":"IL-10 with IL-12 and TNF-\u03b1 attenuates CXCL8\/CXCR1 axis in S. aureus-infected macrophages","volume":"120","author":"Dutta","year":"2023","journal-title":"Int. Immunopharmacol."},{"key":"ref_28","doi-asserted-by":"crossref","unstructured":"Aizaz, M., Khan, A., Khan, F., Khan, M., Musad Saleh, E.A., Nisar, M., and Baran, N. (2023). The cross-talk between macrophages and tumor cells as a target for cancer treatment. Front. Oncol., 13.","DOI":"10.3389\/fonc.2023.1259034"}],"container-title":["Medical Sciences"],"original-title":[],"language":"en","link":[{"URL":"https:\/\/www.mdpi.com\/2076-3271\/13\/3\/129\/pdf","content-type":"unspecified","content-version":"vor","intended-application":"similarity-checking"}],"deposited":{"date-parts":[[2025,10,9]],"date-time":"2025-10-09T18:27:08Z","timestamp":1760034428000},"score":1,"resource":{"primary":{"URL":"https:\/\/www.mdpi.com\/2076-3271\/13\/3\/129"}},"subtitle":[],"short-title":[],"issued":{"date-parts":[[2025,8,14]]},"references-count":28,"journal-issue":{"issue":"3","published-online":{"date-parts":[[2025,9]]}},"alternative-id":["medsci13030129"],"URL":"https:\/\/doi.org\/10.3390\/medsci13030129","relation":{},"ISSN":["2076-3271"],"issn-type":[{"value":"2076-3271","type":"electronic"}],"subject":[],"published":{"date-parts":[[2025,8,14]]}}}