{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,10,12]],"date-time":"2026-10-12T19:21:55Z","timestamp":1791832915035,"version":"4.3.4"},"reference-count":48,"publisher":"MDPI AG","issue":"1","license":[{"start":{"date-parts":[[2020,12,29]],"date-time":"2020-12-29T00:00:00Z","timestamp":1609200000000},"content-version":"vor","delay-in-days":0,"URL":"https:\/\/creativecommons.org\/licenses\/by\/4.0\/"}],"content-domain":{"domain":[],"crossmark-restriction":false},"short-container-title":["IJMS"],"abstract":"<jats:p>For the development and evaluation of new head and neck squamous cell carcinoma (HNSCC) therapeutics, suitable, well-characterized animal models are needed. Thus, by analyzing orthotopic versus subcutaneous models of HNSCC in immunocompetent mice, we evaluated the existence of adenosine-related immunosuppressive B- and T lymphocyte populations within the tumor microenvironment (TME). Applying the SCC VII model for the induction of HNSCC in immunocompetent C3H\/HeN mice, the cellular TME was characterized after tumor initiation over time by flow cytometry. The TME in orthotopic grown tumors revealed a larger population of tumor-infiltrating lymphocytes (TIL) with more B cells and CD4+ T cells than the subcutaneously grown tumors. Immune cell populations in the blood and bone marrow showed a rather distinct reaction toward tumor induction and tumor location compared to the spleen, lymph nodes, or thymus. In addition, large numbers of immunosuppressive B- and T cells were identified within the TME but also in secondary lymphoid organs, independently of the tumor initiation site. The altered immunogenic TME may influence the response to any treatment attempt. Moreover, when analyzing the TME and other lymphoid organs of tumor-bearing mice, we observed conditions reflecting largely those of patients suffering from HNSCC suggesting the C3H\/HeN mouse model as a suitable tool for studies aiming to target immunosuppression to improve anti-cancer therapies.<\/jats:p>","DOI":"10.3390\/ijms22010247","type":"journal-article","created":{"date-parts":[[2020,12,29]],"date-time":"2020-12-29T10:04:46Z","timestamp":1609236286000},"page":"247","update-policy":"https:\/\/doi.org\/10.3390\/mdpi_crossmark_policy","source":"Crossref","is-referenced-by-count":21,"title":["Characterization and Differentiation of the Tumor Microenvironment (TME) of Orthotopic and Subcutaneously Grown Head and Neck Squamous Cell Carcinoma (HNSCC) in Immunocompetent Mice"],"prefix":"10.3390","volume":"22","author":[{"given":"Matthias","family":"Brand","sequence":"first","affiliation":[{"name":"Department of Otorhinolaryngology, University Hospital Ulm, Frauensteige 12, 89075 Ulm, Germany"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0001-6732-7137","authenticated-orcid":false,"given":"Simon","family":"Laban","sequence":"additional","affiliation":[{"name":"Department of Otorhinolaryngology, University Hospital Ulm, Frauensteige 12, 89075 Ulm, Germany"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Marie-Nicole","family":"Theodoraki","sequence":"additional","affiliation":[{"name":"Department of Otorhinolaryngology, University Hospital Ulm, Frauensteige 12, 89075 Ulm, Germany"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Johannes","family":"Doescher","sequence":"additional","affiliation":[{"name":"Department of Otorhinolaryngology, University Hospital Ulm, Frauensteige 12, 89075 Ulm, Germany"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Thomas K.","family":"Hoffmann","sequence":"additional","affiliation":[{"name":"Department of Otorhinolaryngology, University Hospital Ulm, Frauensteige 12, 89075 Ulm, Germany"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-1065-8709","authenticated-orcid":false,"given":"Patrick J.","family":"Schuler","sequence":"additional","affiliation":[{"name":"Department of Otorhinolaryngology, University Hospital Ulm, Frauensteige 12, 89075 Ulm, Germany"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-6456-5508","authenticated-orcid":false,"given":"Cornelia","family":"Brunner","sequence":"additional","affiliation":[{"name":"Department of Otorhinolaryngology, University Hospital Ulm, Frauensteige 12, 89075 Ulm, Germany"}],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"1968","published-online":{"date-parts":[[2020,12,29]]},"reference":[{"key":"ref_1","doi-asserted-by":"crossref","first-page":"67","DOI":"10.1016\/j.oraloncology.2019.01.018","article-title":"The effects of checkpoint inhibition on head and neck squamous cell carcinoma: A systematic review","volume":"90","author":"Ghanizada","year":"2019","journal-title":"Oral Oncol."},{"key":"ref_2","doi-asserted-by":"crossref","first-page":"309","DOI":"10.1016\/j.oraloncology.2008.06.002","article-title":"Global epidemiology of oral and oropharyngeal cancer","volume":"45","author":"Warnakulasuriya","year":"2009","journal-title":"Oral Oncol."},{"key":"ref_3","doi-asserted-by":"crossref","first-page":"489","DOI":"10.4065\/83.4.489","article-title":"Head and neck cancer: Changing epidemiology, diagnosis, and treatment","volume":"Volume 83","author":"Marur","year":"2008","journal-title":"Proceedings of the Mayo Clinic Proceedings"},{"key":"ref_4","doi-asserted-by":"crossref","first-page":"52","DOI":"10.1007\/s12026-013-8462-3","article-title":"The immune system and head and neck squamous cell carcinoma: From carcinogenesis to new therapeutic opportunities","volume":"57","author":"Freiser","year":"2013","journal-title":"Immunol. Res."},{"key":"ref_5","doi-asserted-by":"crossref","first-page":"1","DOI":"10.1186\/s40425-016-0145-x","article-title":"B cell regulation of the anti-tumor response and role in carcinogenesis","volume":"4","author":"Schwartz","year":"2016","journal-title":"J. Immunother. Cancer"},{"key":"ref_6","doi-asserted-by":"crossref","first-page":"1856","DOI":"10.1056\/NEJMoa1602252","article-title":"Nivolumab for recurrent squamous-cell carcinoma of the head and neck","volume":"375","author":"Ferris","year":"2016","journal-title":"N. Engl. J. Med."},{"key":"ref_7","doi-asserted-by":"crossref","first-page":"113","DOI":"10.1007\/s00005-017-0492-4","article-title":"Myeloid-Derived Suppressor Cells in the Tumor Microenvironment: Current Knowledge and Future Perspectives","volume":"66","author":"Zuazo","year":"2018","journal-title":"Arch. Immunol. Ther. Exp. (Warsz.)"},{"key":"ref_8","doi-asserted-by":"crossref","first-page":"103713","DOI":"10.1016\/j.cellimm.2017.10.013","article-title":"Targeting myeloid cells in the tumor sustaining microenvironment","volume":"343","author":"Schupp","year":"2019","journal-title":"Cell. Immunol."},{"key":"ref_9","doi-asserted-by":"crossref","first-page":"1205","DOI":"10.1007\/s00262-020-02535-6","article-title":"Adenosine-producing regulatory B cells in head and neck cancer","volume":"69","author":"Jeske","year":"2020","journal-title":"Cancer Immunol. Immunother."},{"key":"ref_10","doi-asserted-by":"crossref","first-page":"55","DOI":"10.3342\/ceo.2009.2.2.55","article-title":"Animal models of cancer in the head and neck region","volume":"2","author":"Kim","year":"2009","journal-title":"Clin. Exp. Otorhinolaryngol."},{"key":"ref_11","doi-asserted-by":"crossref","first-page":"199","DOI":"10.1016\/j.canlet.2011.11.027","article-title":"Optimization of an orthotopic murine model of head and neck squamous cell carcinoma in fully immunocompetent mice - Role of toll-like-receptor 4 expressed on host cells","volume":"317","author":"Vahle","year":"2012","journal-title":"Cancer Lett."},{"key":"ref_12","doi-asserted-by":"crossref","first-page":"20","DOI":"10.1001\/archotol.1997.01900010022003","article-title":"A new immunocompetent murine model for oral cancer","volume":"123","author":"Cope","year":"1997","journal-title":"Arch. Otolaryngol.-Head Neck Surg."},{"key":"ref_13","doi-asserted-by":"crossref","first-page":"1263","DOI":"10.1001\/archotol.133.12.1263","article-title":"The effect of altered toll-like receptor 4 signaling on cancer cachexia","volume":"133","author":"Cannon","year":"2007","journal-title":"Arch. Otolaryngol. Head Neck Surg."},{"key":"ref_14","doi-asserted-by":"crossref","first-page":"1473","DOI":"10.1016\/0360-3016(84)90371-7","article-title":"Modification of the effects of continuous low dose rate irradiation by concurrent chemotherapy infusion","volume":"10","author":"Fu","year":"1984","journal-title":"Int. J. Radiat. Oncol. Biol. Phys."},{"key":"ref_15","doi-asserted-by":"crossref","first-page":"51","DOI":"10.1016\/j.oraloncology.2016.12.010","article-title":"Preclinical models in HNSCC: A comprehensive review","volume":"65","author":"Rancoule","year":"2017","journal-title":"Oral Oncol."},{"key":"ref_16","doi-asserted-by":"crossref","first-page":"2111","DOI":"10.1002\/ijc.21694","article-title":"Animal models for the study of squamous cell carcinoma of the upper aerodigestive tract: A historical perspective with review of their utility and limitations. Part A. Chemically-inducedde novo cancer, syngeneic animal models of HNSCC, animal models of transplanted xenogeneic human tumors","volume":"118","author":"Smith","year":"2006","journal-title":"Int. J. Cancer"},{"key":"ref_17","doi-asserted-by":"crossref","first-page":"899","DOI":"10.1002\/hed.1130","article-title":"Characterization of a spontaneously arising murine squamous cell carcinoma (SCC VII) as a prerequisite for head and neck cancer immunotherapy","volume":"23","author":"Khurana","year":"2001","journal-title":"Head Neck"},{"key":"ref_18","doi-asserted-by":"crossref","first-page":"13132","DOI":"10.1073\/pnas.0605251103","article-title":"A2A adenosine receptor protects tumors from antitumor T cells","volume":"103","author":"Ohta","year":"2006","journal-title":"Proc. Natl. Acad. Sci. USA"},{"key":"ref_19","doi-asserted-by":"crossref","first-page":"4977","DOI":"10.4049\/jimmunol.1001323","article-title":"CD20 + B Cells: The Other Tumor-Infiltrating Lymphocytes","volume":"185","author":"Nelson","year":"2010","journal-title":"J. Immunol."},{"key":"ref_20","doi-asserted-by":"crossref","first-page":"26","DOI":"10.1016\/j.smim.2007.12.004","article-title":"Ectopic lymphoid tissues and local immunity","volume":"20","author":"Carragher","year":"2008","journal-title":"Semin. Immunol."},{"key":"ref_21","doi-asserted-by":"crossref","first-page":"4006","DOI":"10.4049\/jimmunol.0903009","article-title":"B Cells Are Required for Optimal CD4 + and CD8 + T Cell Tumor Immunity: Therapeutic B Cell Depletion Enhances B16 Melanoma Growth in Mice","volume":"184","author":"DiLillo","year":"2010","journal-title":"J. Immunol."},{"key":"ref_22","doi-asserted-by":"crossref","first-page":"128","DOI":"10.1158\/0008-5472.CAN-12-2606","article-title":"PD-1-expressing tumor-infiltrating T cells are a favorable prognostic biomarker in HPV-Associated head and neck cancer","volume":"73","author":"Badoual","year":"2013","journal-title":"Cancer Res."},{"key":"ref_23","doi-asserted-by":"crossref","first-page":"465","DOI":"10.1158\/1078-0432.CCR-05-1886","article-title":"Prognostic value of tumor-infiltrating CD4+ T-cell subpopulations in head and neck cancers","volume":"12","author":"Badoual","year":"2006","journal-title":"Clin. Cancer Res."},{"key":"ref_24","first-page":"3675","article-title":"p53110-124-specific human CD4+ T-helper cells enhance in vitro generation and antitumor function of tumor-reactive CD8+ T cells","volume":"63","author":"Chikamatsu","year":"2003","journal-title":"Cancer Res."},{"key":"ref_25","doi-asserted-by":"crossref","first-page":"393","DOI":"10.1007\/s00262-016-1805-x","article-title":"A beneficial tumor microenvironment in oropharyngeal squamous cell carcinoma is characterized by a high T cell and low IL-17+ cell frequency","volume":"65","author":"Punt","year":"2016","journal-title":"Cancer Immunol. Immunother."},{"key":"ref_26","doi-asserted-by":"crossref","first-page":"3706","DOI":"10.1158\/1078-0432.CCR-07-5126","article-title":"T regulatory type 1 cells in squamous cell carcinoma of the head and neck: Mechanisms of suppression and expansion in advanced disease","volume":"14","author":"Bergmann","year":"2008","journal-title":"Clin. Cancer Res."},{"key":"ref_27","doi-asserted-by":"crossref","first-page":"6780","DOI":"10.4049\/jimmunol.177.10.6780","article-title":"T Regulatory and Primed Uncommitted CD4 T Cells Express CD73, Which Suppresses Effector CD4 T Cells by Converting 5\u2032-Adenosine Monophosphate to Adenosine","volume":"177","author":"Kobie","year":"2006","journal-title":"J. Immunol."},{"key":"ref_28","first-page":"2290","article-title":"Experimental cancer cachexia induced by transplantable colon 26 adenocarcinoma in mice","volume":"50","author":"Tanaka","year":"1990","journal-title":"Cancer Res."},{"key":"ref_29","doi-asserted-by":"crossref","first-page":"1650","DOI":"10.1056\/NEJM198612253152606","article-title":"Tumors: Wounds That Do Not Heal","volume":"315","author":"Flier","year":"1986","journal-title":"N. Engl. J. Med."},{"key":"ref_30","doi-asserted-by":"crossref","first-page":"654","DOI":"10.1016\/j.jaci.2016.07.006","article-title":"Role of regulatory B cells in immune tolerance to allergens and beyond","volume":"138","author":"Stanic","year":"2016","journal-title":"J. Allergy Clin. Immunol."},{"key":"ref_31","doi-asserted-by":"crossref","first-page":"5541","DOI":"10.4049\/jimmunol.175.8.5541","article-title":"Inhibition of NK Cell Activity through TGF-\u03b21 by Down-Regulation of NKG2D in a Murine Model of Head and Neck Cancer","volume":"175","author":"Dasgupta","year":"2005","journal-title":"J. Immunol."},{"key":"ref_32","doi-asserted-by":"crossref","first-page":"2523","DOI":"10.1158\/0008-5472.CAN-08-2793","article-title":"Interleukin-27 activates natural killer cells and suppresses NK-resistant head and neck squamous cell carcinoma through inducing antibody-dependent cellular cytotoxicity","volume":"69","author":"Matsui","year":"2009","journal-title":"Cancer Res."},{"key":"ref_33","doi-asserted-by":"crossref","first-page":"236","DOI":"10.1016\/j.vascn.2010.12.001","article-title":"Development of a murine model of lymph node metastases suitable for immunotoxicity studies","volume":"63","author":"Egenolf","year":"2011","journal-title":"J. Pharmacol. Toxicol. Methods"},{"key":"ref_34","doi-asserted-by":"crossref","first-page":"1565","DOI":"10.1126\/science.1203486","article-title":"Cancer immunoediting: Integrating immunity\u2019s roles in cancer suppression and promotion","volume":"331","author":"Schreiber","year":"2011","journal-title":"Science"},{"key":"ref_35","doi-asserted-by":"crossref","first-page":"3755","DOI":"10.1158\/1078-0432.CCR-04-0054","article-title":"Decreased absolute counts of T lymphocyte subsets and their relation to disease in squamous cell carcinoma of the head and neck","volume":"10","author":"Kuss","year":"2004","journal-title":"Clin. Cancer Res."},{"key":"ref_36","first-page":"6811","article-title":"Tumor-induced apoptosis of T cells: Amplification by a mitochondrial cascade","volume":"60","author":"Gastman","year":"2000","journal-title":"Cancer Res."},{"key":"ref_37","doi-asserted-by":"crossref","first-page":"3105","DOI":"10.1158\/0008-5472.CAN-08-3838","article-title":"Triggering of toll-like receptor 4 expressed on human head and neck squamous cell carcinoma promotes tumor development and protects the tumor from immune attack","volume":"69","author":"Szczepanski","year":"2009","journal-title":"Cancer Res."},{"key":"ref_38","doi-asserted-by":"crossref","first-page":"1241","DOI":"10.1517\/14712598.2013.810716","article-title":"Immune alterations and immunotherapy prospects in head and neck cancer","volume":"13","author":"Varilla","year":"2013","journal-title":"Expert Opin. Biol. Ther."},{"key":"ref_39","doi-asserted-by":"crossref","first-page":"309","DOI":"10.1016\/j.otohns.2004.04.018","article-title":"Prognostic significance of Langerhans cell infiltration in cancer of the larynx","volume":"132","author":"Yilmaz","year":"2005","journal-title":"Otolaryngol. Head Neck Surg."},{"key":"ref_40","doi-asserted-by":"crossref","first-page":"259","DOI":"10.1111\/j.0959-9673.2004.00361.x","article-title":"Correlation of c-erbB-2 and S-100 expression with the malignancy grading and anatomical site in oral squamous cell carcinoma","volume":"84","author":"Albuquerque","year":"2003","journal-title":"Int. J. Exp. Pathol."},{"key":"ref_41","doi-asserted-by":"crossref","first-page":"273","DOI":"10.1016\/j.smim.2005.05.009","article-title":"Cutaneous dendritic cells","volume":"17","author":"Valladeau","year":"2005","journal-title":"Semin. Immunol."},{"key":"ref_42","doi-asserted-by":"crossref","first-page":"481","DOI":"10.1016\/S0092-8674(00)00139-2","article-title":"MMP-9 supplied by bone marrow-derived cells contributes to skin carcinogenesis","volume":"103","author":"Coussens","year":"2000","journal-title":"Cell"},{"key":"ref_43","first-page":"1128","article-title":"Vascular endothelial growth factor in squamous cell head and neck carcinoma: Expression and prognostic significance","volume":"10","author":"Salven","year":"1997","journal-title":"Mod. Pathol."},{"key":"ref_44","doi-asserted-by":"crossref","unstructured":"Pretscher, D., Distel, L.V., Grabenbauer, G.G., Wittlinger, M., Buettner, M., and Niedobitek, G. (2009). Distribution of immune cells in head and neck cancer: CD8+T-cells and CD20+B-cells in metastatic lymph nodes are associated with favourable outcome in patients with oro- and hypopharyngeal carcinoma. BMC Cancer, 9.","DOI":"10.1186\/1471-2407-9-292"},{"key":"ref_45","doi-asserted-by":"crossref","first-page":"e167","DOI":"10.1016\/j.oraloncology.2009.05.640","article-title":"Tumour infiltrating lymphocytes in squamous cell carcinoma of the oro- and hypopharynx: Prognostic impact may depend on type of treatment and stage of disease","volume":"45","author":"Distel","year":"2009","journal-title":"Oral Oncol."},{"key":"ref_46","doi-asserted-by":"crossref","first-page":"1","DOI":"10.1186\/1476-4598-9-4","article-title":"Different subsets of tumor infiltrating lymphocytes correlate with NPC progression in different ways","volume":"9","author":"Zhang","year":"2010","journal-title":"Mol. Cancer"},{"key":"ref_47","doi-asserted-by":"crossref","first-page":"270","DOI":"10.1158\/2159-8290.CD-15-0827","article-title":"Bruton tyrosine kinase\u2013Dependent immune cell cross-talk drives pancreas cancer","volume":"6","author":"Gunderson","year":"2016","journal-title":"Cancer Discov."},{"key":"ref_48","doi-asserted-by":"crossref","first-page":"1287","DOI":"10.1084\/jem.188.7.1287","article-title":"Involvement of guanosine triphosphatases and phospholipase C-\u03b32 in extracellular signal-regulated kinase, c-Jun NH2-terminal kinase, and p38 mitogen-activated protein kinase activation by the B cell antigen receptor","volume":"188","author":"Hashimoto","year":"1998","journal-title":"J. Exp. Med."}],"container-title":["International Journal of Molecular Sciences"],"original-title":[],"language":"en","link":[{"URL":"https:\/\/www.mdpi.com\/1422-0067\/22\/1\/247\/pdf","content-type":"unspecified","content-version":"vor","intended-application":"similarity-checking"}],"deposited":{"date-parts":[[2025,10,11]],"date-time":"2025-10-11T10:47:15Z","timestamp":1760179635000},"score":1,"resource":{"primary":{"URL":"https:\/\/www.mdpi.com\/1422-0067\/22\/1\/247"}},"subtitle":[],"short-title":[],"issued":{"date-parts":[[2020,12,29]]},"references-count":48,"journal-issue":{"issue":"1","published-online":{"date-parts":[[2021,1]]}},"alternative-id":["ijms22010247"],"URL":"https:\/\/doi.org\/10.3390\/ijms22010247","relation":{},"ISSN":["1422-0067"],"issn-type":[{"value":"1422-0067","type":"electronic"}],"subject":[],"published":{"date-parts":[[2020,12,29]]}}}