IntroductionHead and neck squamous cell carcinoma (HNSCC) strongly suppresses the immune system, resulting in increased metastasis and recurrent disease. Chemotherapy is part of the multimodal treatment but may further immunosuppression. Recently, we demonstrated that regulatory B cells (Breg), defined as CD19+CD39+CD73+ B cells, play a significant role in the production of immunosuppressive, extracellular adenosine (ADO). Here, we tested the influence of chemotherapy on Breg function.ResultsIn HNSCC patients, Breg were diminished in absolute number and frequency after chemotherapy (paired samples). Chemotherapeutic drugs had variable effects; while platinum-based chemotherapy decreased the expression of CD39, methotrexate led to a functional increase in CD39 expression and increased production of immunosuppressive ADO. These findings were confirmed in a second patient cohort. Surface expression of CD39 correlated strongly with the production of ADO as measured by mass spectrometry.ConclusionsPlatinum-based anti-tumor-therapy reduces the number of adenosine-producing B cells and, consequently, potential immunosuppression within the tumor environment. Breg function in terms of ADO production and their potential capacity to suppress CD4+ T cells are promoted by methotrexate treatment amplifying anti-inflammatory therapeutic effects. Our results add to the understanding of how chemotherapeutic drugs can influence the human immune system and may therefore help to orchestrate standard oncologic therapy with new immune modulating approaches.MethodsMononuclear cells were collected prospectively from HNSCC patients before and after chemotherapy (n = 18), from healthy donors (n = 20), and an additional cohort sampled several months after chemotherapy (n = 14). Frequency, phenotype, and function of Breg were determined by multicolor flow cytometry, ATP luminescence assay as well as mass spectrometry measuring 5′-AMP, ADO, and inosine. Isolated B cells were incubated with chemotherapeutic drugs (cisplatin, methotrexate, paclitaxel, 5-fluorouracil) in vitro for functional studies.
Background Multiple mechanisms of immunosuppression have been identified in the tumor microenvironment including regulatory B cells (B reg). Recently, we have shown that B reg suppress T cell function by production of adenosine (ADO). However, the autocrine effect of ADO on B cells and the role of B reg in head and neck cancer remains unclear. Methods Blood (n = 42) and tumor tissue (n = 39) of head and neck cancer patients and healthy donors (n = 60) were analyzed by FACS. The effect of ADO on phenotype, intracellular signaling pathways, Ca 2+ influx and ADO production was analyzed in B reg and effector B cells (B eff) by FACS, luminescence and mass spectrometry. The blockage of the ADO receptor A 2A was analyzed in a murine head and neck cancer model. Results ADO-producing B reg were found in tumor tissue and peripheral blood. ADO inhibited the intracellular Bruton's tyrosine kinase (BTK) and Ca 2+ influx only in B eff. The inhibition of BTK by ibrutinib mimicked the effect of ADO, and ibrutinib reduced the production of ADO by downregulation of CD39 in vitro. The inhibition of ADO receptor A 2A significantly reduced tumor mass and increased B cell infiltration, in vivo. Conclusion Our data demonstrate the presence of a novel ADO-producing B reg population within the tumor microenvironment in mice and humans. A new model is proposed on how ADO-producing B reg can influence the function of B eff cells in healthy donors and cancer patients. Thus, the modulation of the ADO pathway in B cells may serve as a therapeutic approach for cancer patients. Keywords Regulatory B cells • Adenosine • BTK • Head and neck cancer • ADORA 2A Abbreviations ADO Adenosine ADORA Adenosine receptor A BCR B cell receptor B reg Regulatory B cells BTK Bruton's tyrosine kinase GAPDH Glyceraldehyde 3-phosphate dehydrogenase G6PD Glucose-6-phosphate dehydrogenase HNSCC Head and neck squamous cell carcinoma PBG-D Porphobilinogen deaminase RPL13A Ribosomal protein L13a SCC Squamous cell carcinoma TBP TATA-box-binding protein T reg Regulatory T cells Electronic supplementary material The online version of this article (
BackgroundReliable on site classification of resected tumor specimens remains a challenge. Implementation of high-resolution confocal laser endoscopic techniques (CLEs) during fluorescence-guided brain tumor surgery is a new tool for intraoperative tumor tissue visualization. To overcome observer dependent errors, we aimed to predict tumor type by applying a deep learning model to image data obtained by CLE.MethodsHuman brain tumor specimens from 25 patients with brain metastasis, glioblastoma, and meningioma were evaluated within this study. In addition to routine histopathological analysis, tissue samples were stained with fluorescein ex vivo and analyzed with CLE. We trained two convolutional neural networks and built a predictive level for the outputs.ResultsMultiple CLE images were obtained from each specimen with a total number of 13,972 fluorescein based images. Test accuracy of 90.9% was achieved after applying a two-class prediction for glioblastomas and brain metastases with an area under the curve (AUC) value of 0.92. For three class predictions, our model achieved a ratio of correct predicted label of 85.8% in the test set, which was confirmed with five-fold cross validation, without definition of confidence. Applying a confidence rate of 0.999 increased the prediction accuracy to 98.6% when images with substantial artifacts were excluded before the analysis. 36.3% of total images met the output criteria.ConclusionsWe trained a residual network model that allows automated, on site analysis of resected tumor specimens based on CLE image datasets. Further in vivo studies are required to assess the clinical benefit CLE can have.
Objective: The subarachnoid haemorrhage (SAH) outcome tool (SAHOT) is the first SAH-specific patient reported outcome measure, and was developed in the UK. We aimed to validate the SAHOT outside the UK, and therefore endeavored to adapt the SAHOT into German and to test its psychometric properties. Methods: We adapted and pilot tested the German version. We applied the SAHOT, Quality of Life after Brain Injury, Hospital Anxiety and Depression Scale, and EuroQol questionnaires in a cohort of 89 patients with spontaneous SAH after discharge. We assessed internal consistency by Cronbach’s α, test-retest reliability by intraclass correlation, and validity by Pearson correlations with established measures. Sensitivity to change was evaluated following neurorehabilitation by effect sizes. Results: The translation of SAHOT resulted in a German version that is semantically and conceptually equivalent to the English version. Internal consistency was good regarding the physical domain (α = 0.83) and excellent for the other domains (α = 0.92–0.93). Test–retest reliability indicated a high level of stability with an intraclass correlation of 0.85 (95% CI: 0.83–0.86). All domains correlated moderately or strongly with established measures ( r = 0.41–0.74; p < 0.01). SAHOT total scores showed moderate sensitivity to change (Cohen’s d = −0.68), while mRS and GOSE showed no significant sensitivity to change. Conclusion: The SAHOT can be adapted to other health care systems and societies than the UK. The German version of the SAHOT is a reliable and valid instrument, and can be used in future clinical studies and individual assessment after spontaneous SAH.
Purpose: Head and Neck Squamous Cell Carcinoma (HNSCC) strongly affects the immune system resulting in profound immune defects. These are associated with increased metastasis and recurrent disease. Standard treatment options include among others various regimens of chemotherapy. However, besides the anti-neoplastic effects of chemotherapy, adverse immune modulations are described. Recently, we demonstrated that regulatory B cells (Breg) play a significant role in the production of extracellular adenosine (ADO) using ectonucleotidases CD39 and CD73, which is known to be highly immunosuppressive. Here, we tested the influence of chemotherapy on the production of ADO by Breg. Experimental Design: Mononuclear cells were isolated from peripheral blood collected from 30 healthy donors and from patients with HNSCC before (n=30) and after chemotherapy (n=30). The number, frequency and phenotype of Breg was determined by multicolor flow cytometry. Next, B cells were isolated from healthy donors by CD19+ magnetic beads, stimulated (CD40L, IL-4, hemagglutinin) and incubated for 7 days with physiologic concentrations of various chemotherapeutic drugs (cisplatin, methotrexate, paclitaxel, 5-fluorouracil, cetuximab) for 7 days. Effects on Breg proliferation were determined by CFSE proliferation assay. Production of extracellular ADO before and after chemotherapy was determined by flow cytometry, ATP luminescence assay as well as highly sensitive mass spectrometry measuring 5'AMP, ADO and inosine. Long-term effects were described analyzing Breg from patients who had received chemotherapy several years ago. Results: Breg were numerically diminished in untreated HNSCC patients. However, subsequently to a strong numerical decrease of CD4+ T cells during chemotherapy, the frequency of B cells was increased. Chemotherapeutic drugs had variable effects in vitro and in vivo: While cisplatin, paclitaxel and 5-fluorouracil decreased expression of CD39, it increased under methotrexate. Importantly, hydrolysis of ATP to exogenous ADO correlated strongly with CD39 expression levels (p < 0.05). CD73 expression levels were increased by all drugs, except for cetuximab, which showed no effect on phenotype or function. Conclusion: Highly suppressive Breg increase in frequency after chemotherapy and could be responsible for suppression of antitumor immune responses and recurrent disease in HNSCC. B cell function in terms of ADO production and their capacity to suppress CD4+ T cells is compromised by various chemotherapeutic drugs in vitro and in vivo amplifying therapeutic benefits. Citation Format: Andreas Ziebart, Ulrich Huber, Silke Schwarz, Simon Laban, Thomas K. Hoffmann, Patrick Schuler. The influence of chemotherapy on adenosine-producing B cells in patients with head and neck cancer. [abstract]. In: Proceedings of the AACR Special Conference: Tumor Immunology and Immunotherapy: A New Chapter; December 1-4, 2014; Orlando, FL. Philadelphia (PA): AACR; Cancer Immunol Res 2015;3(10 Suppl):Abstract nr B50.
scite is a Brooklyn-based organization that helps researchers better discover and understand research articles through Smart Citations–citations that display the context of the citation and describe whether the article provides supporting or contrasting evidence. scite is used by students and researchers from around the world and is funded in part by the National Science Foundation and the National Institute on Drug Abuse of the National Institutes of Health.
hi@scite.ai
10624 S. Eastern Ave., Ste. A-614
Henderson, NV 89052, USA
Copyright © 2024 scite LLC. All rights reserved.
Made with 💙 for researchers
Part of the Research Solutions Family.