{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,3,2]],"date-time":"2026-03-02T13:04:32Z","timestamp":1772456672543,"version":"3.50.1"},"reference-count":56,"publisher":"Frontiers Media SA","license":[{"start":{"date-parts":[[2023,3,23]],"date-time":"2023-03-23T00:00:00Z","timestamp":1679529600000},"content-version":"vor","delay-in-days":0,"URL":"https:\/\/creativecommons.org\/licenses\/by\/4.0\/"}],"funder":[{"DOI":"10.13039\/501100004281","name":"Narodowe Centrum Nauki","doi-asserted-by":"publisher","award":["2015\/19\/N\/NZ6\/02908"],"award-info":[{"award-number":["2015\/19\/N\/NZ6\/02908"]}],"id":[{"id":"10.13039\/501100004281","id-type":"DOI","asserted-by":"publisher"}]}],"content-domain":{"domain":["frontiersin.org"],"crossmark-restriction":true},"short-container-title":["Front. Immunol."],"abstract":"<jats:sec><jats:title>Background<\/jats:title><jats:p>Understanding the negative impact of the tumor microenvironment on the creation of an effective immune response has contributed to the development of new therapeutic anti-cancer strategies. One such solution is combined therapy consisting of chemotherapeutic administration followed by dendritic cell (DC)-based vaccines. The use of cytostatic leads to the elimination of cancer cells, but can also modulate the tumor milieu. Moreover, great efforts are being made to increase the therapeutic outcome of immunotherapy, e.g. by enhancing the ability of DCs to generate an efficient immune response, even in the presence of immunosuppressive cytokines such as IL-10. The study aimed to determine the effectiveness of combined therapy with chemotherapeutic with immunomodulatory potential \u2013 HES-MTX nanoconjugate (composed of methotrexate (MTX) and hydroxyethyl starch (HES)) and DCs with downregulated expression of IL-10 receptor stimulated with tumor antigens (DC\/shIL-10R\/TAg) applied in MC38 murine colon carcinoma model.<\/jats:p><\/jats:sec><jats:sec><jats:title>Methods<\/jats:title><jats:p>With the use of lentiviral vectors the DCs with decreased expression of IL-10R were obtained and characterized. During <jats:italic>in vivo<\/jats:italic> studies MC38-tumor bearing mice received MTX or HES-MTX nanoconjugate as a sole treatment or combined with DC-based immunotherapy containing unmodified DCs or DCs transduced with shRNA against IL-10R (or control shRNA sequence). Tumor volume was monitored during the experiment. One week after the last injection of DC-based vaccines, tumor nodules and spleens were dissected for <jats:italic>ex vivo<\/jats:italic> analysis. The changes in the local and systemic anti-tumor immune response were estimated with the use of flow cytometry and ELISA methods.<\/jats:p><\/jats:sec><jats:sec><jats:title>Results and conclusions<\/jats:title><jats:p><jats:italic>In vitro<\/jats:italic> studies showed that the downregulation of IL-10R expression in DCs enhances their ability to activate the specific anti-tumor immune response. The use of HES-MTX nanoconjugate and DC\/shIL-10R\/TAg in the therapy of MC38-tumor bearing mice resulted in the greatest tumor growth inhibition. At the local anti-tumor immune response level a decrease in the infiltration of cells with suppressor activity and an increase in the influx of effector cells into MC38 tumor tissue was observed. These changes were crucial to enhance the effective specific immune response at the systemic level, which was revealed in the greatest cytotoxic activity of spleen cells against MC38 cells.<\/jats:p><\/jats:sec>","DOI":"10.3389\/fimmu.2023.1155377","type":"journal-article","created":{"date-parts":[[2023,3,23]],"date-time":"2023-03-23T04:47:14Z","timestamp":1679546834000},"update-policy":"https:\/\/doi.org\/10.3389\/crossmark-policy","source":"Crossref","is-referenced-by-count":11,"title":["Combined therapy with methotrexate nanoconjugate and dendritic cells with downregulated IL-10R expression modulates the tumor microenvironment and enhances the systemic anti-tumor immune response in MC38 murine colon carcinoma"],"prefix":"10.3389","volume":"14","author":[{"given":"Agnieszka","family":"Szczygie\u0142","sequence":"first","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Katarzyna","family":"W\u0119gierek-Ciura","sequence":"additional","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Anna","family":"Wr\u00f3blewska","sequence":"additional","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Jagoda","family":"Mierzejewska","sequence":"additional","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Joanna","family":"Rossowska","sequence":"additional","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Bo\u017cena","family":"Szermer-Olearnik","sequence":"additional","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Marta","family":"\u015awitalska","sequence":"additional","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Natalia","family":"Anger-G\u00f3ra","sequence":"additional","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Tomasz M.","family":"Goszczy\u0144ski","sequence":"additional","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"El\u017cbieta","family":"Pajtasz-Piasecka","sequence":"additional","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]}],"member":"1965","published-online":{"date-parts":[[2023,3,23]]},"reference":[{"key":"B1","doi-asserted-by":"publisher","DOI":"10.1084\/jem.137.5.1142","article-title":"Identification of a novel cell type in peripheral lymphoid organs of mice. i. morphology, quantitation, tissue distribution","volume":"137","author":"Steinman","year":"1973","journal-title":"J Exp Med"},{"key":"B2","doi-asserted-by":"publisher","first-page":"1","DOI":"10.1146\/annurev-immunol-100311-102839","article-title":"Decisions about dendritic cells: past, present, and future","volume":"30","author":"Steinman","year":"2012","journal-title":"Annu Rev Immunol"},{"key":"B3","doi-asserted-by":"publisher","DOI":"10.3389\/fcell.2021.686544","article-title":"Insights into dendritic cells in cancer immunotherapy: From bench to clinical applications","volume":"9","author":"Salah","year":"2021","journal-title":"Front Cell Dev Biol"},{"key":"B4","doi-asserted-by":"publisher","DOI":"10.3389\/fonc.2021.730824","article-title":"Recent advances in experimental dendritic cell vaccines for cancer","volume":"11","author":"Filin","year":"2021","journal-title":"Front Oncol"},{"key":"B5","doi-asserted-by":"publisher","first-page":"179","DOI":"10.1016\/bs.ircmb.2019.07.008","article-title":"Interplay between dendritic cells and cancer cells","volume":"348","author":"Martinek","year":"2019","journal-title":"Int Rev Cell Mol Biol"},{"key":"B6","doi-asserted-by":"publisher","first-page":"289","DOI":"10.2217\/imt-2016-0135","article-title":"Tumor-associated macrophages: Implications in cancer immunotherapy","volume":"9","author":"Petty","year":"2017","journal-title":"Immunotherapy"},{"key":"B7","doi-asserted-by":"publisher","DOI":"10.3389\/fimmu.2011.00029","article-title":"IL-10 immunomodulation of myeloid cells regulates a murine model of ovarian cancer","volume":"2","author":"Hart","year":"2011","journal-title":"Front Immunol"},{"key":"B8","doi-asserted-by":"publisher","DOI":"10.1111\/cas.14069","article-title":"(Treg) cells in cancer: Can treg cells be a new therapeutic target","volume":"110","author":"Ohue","year":"2019","journal-title":"Cancer Sci"},{"key":"B9","doi-asserted-by":"publisher","DOI":"10.4049\/jimmunol.166.7.4312","article-title":"Regulatory activity of autocrine IL-10 on dendritic cell functions","volume":"166","author":"Corinti","year":"2001","journal-title":"J Immunol"},{"key":"B10","doi-asserted-by":"publisher","DOI":"10.1182\/blood.V99.7.2468","article-title":"CD4(+) and CD8(+) anergic T cells induced by interleukin-10-treated human dendritic cells display antigen-specific suppressor activity","volume":"99","author":"Steinbrink","year":"2002","journal-title":"Blood"},{"key":"B11","doi-asserted-by":"publisher","DOI":"10.1159\/000094255","article-title":"Interleukin-10-treated dendritic cells do not inhibit Th2 immune responses in ovalbumin\/alum-sensitized mice","volume":"141","author":"Bellinghausen","year":"2006","journal-title":"Int Arch Allergy Immunol"},{"key":"B12","doi-asserted-by":"publisher","first-page":"2113","DOI":"10.1038\/s41467-018-04524-x","article-title":"Induction of anergic or regulatory tumor-specific CD4+ T cells in the tumor-draining lymph node","volume":"9","author":"Alonso","year":"2018","journal-title":"Nat Commun"},{"key":"B13","doi-asserted-by":"publisher","first-page":"e0139598","DOI":"10.1371\/journal.pone.0139598","article-title":"Serum IL-10 predicts worse outcome in cancer patients: A meta-analysis","volume":"10","author":"Zhao","year":"2015","journal-title":"PloS One"},{"key":"B14","doi-asserted-by":"publisher","DOI":"10.4103\/0973-1482.199448","article-title":"Dual association of serum interleukin-10 levels with colorectal cancer","volume":"13","author":"Abtahi","year":"2017","journal-title":"J Cancer Res Ther"},{"key":"B15","doi-asserted-by":"publisher","DOI":"10.3892\/ol.2019.10338","article-title":"Predictive value of IL-18 and IL-10 in the prognosis of patients with colorectal cancer","volume":"18","author":"Li","year":"2019","journal-title":"Oncol Lett"},{"key":"B16","doi-asserted-by":"publisher","first-page":"120","DOI":"10.1186\/1479-5876-11-120","article-title":"Comparative analysis of cancer vaccine settings for the selection of an effective protocol in mice","volume":"11","author":"Kalli","year":"2013","journal-title":"J Transl Med"},{"key":"B17","doi-asserted-by":"publisher","DOI":"10.1016\/j.imbio.2014.10.009","article-title":"Temporary elimination of IL-10 enhanced the effectiveness of cyclophosphamide and BMDC-based therapy by decrease of the suppressor activity of MDSCs and activation of antitumour immune response","volume":"220","author":"Rossowska","year":"2015","journal-title":"Immunobiology"},{"key":"B18","doi-asserted-by":"publisher","DOI":"10.1097\/CJI.0000000000000212","article-title":"Anti-IL-10-mediated enhancement of antitumor efficacy of a dendritic cell-targeting MIP3\u03b1-gp100 vaccine in the B16F10 mouse melanoma model is dependent on type I interferons","volume":"41","author":"Gordy","year":"2018","journal-title":"J Immunother"},{"key":"B19","doi-asserted-by":"publisher","DOI":"10.1016\/j.ccell.2014.09.006","article-title":"Macrophage IL-10 blocks CD8+ T cell-dependent responses to chemotherapy by suppressing IL-12 expression in intratumoral dendritic cells","volume":"26","author":"Ruffell","year":"2014","journal-title":"Cancer Cell"},{"key":"B20","doi-asserted-by":"publisher","DOI":"10.1016\/j.cellimm.2014.06.002","article-title":"IL-10 signalling blockade at the time of immunization inhibits human papillomavirus 16 E7 transformed TC-1 tumour cells growth in mice","volume":"290","author":"Chen","year":"2014","journal-title":"Cell Immunol"},{"key":"B21","doi-asserted-by":"publisher","DOI":"10.1080\/2162402X.2015.1075113","article-title":"Vaccine-induced but not tumor-derived interleukin-10 dictates the efficacy of interleukin-10 blockade in therapeutic vaccination","volume":"5","author":"Llopiz","year":"2016","journal-title":"Oncoimmunology"},{"key":"B22","doi-asserted-by":"publisher","DOI":"10.1111\/j.1365-2249.2011.04410.x","article-title":"Blocking the immunosuppressive axis with small interfering RNA targeting interleukin (IL)-10 receptor enhances dendritic cell-based vaccine potency","volume":"165","author":"Kim","year":"2011","journal-title":"Clin Exp Immunol"},{"key":"B23","doi-asserted-by":"publisher","DOI":"10.3389\/fimmu.2021.690817","article-title":"Impact of interleukin 10 deficiency on intestinal epithelium responses to inflammatory signals","volume":"12","author":"Papoutsopoulou","year":"2021","journal-title":"Front Immunol"},{"key":"B24","doi-asserted-by":"publisher","first-page":"126","DOI":"10.1186\/s13046-018-0799-y","article-title":"Reprogramming the murine colon cancer microenvironment using lentivectors encoding shRNA against IL-10 as a component of a potent DC-based chemoimmunotherapy","volume":"37","author":"Rossowska","year":"2018","journal-title":"J Exp Clin Cancer Res"},{"key":"B25","doi-asserted-by":"publisher","DOI":"10.1080\/21645515.2019.1701913","article-title":"Suppression of TGF-\u03b2 and IL-10 receptors on self-differentiated dendritic cells by short-hairpin RNAs enhanced activation of effector T-cells against cholangiocarcinoma cells","volume":"16","author":"Thepmalee","year":"2020","journal-title":"Hum Vaccin Immunother"},{"key":"B26","doi-asserted-by":"publisher","DOI":"10.1111\/cei.12620","article-title":"The siRNA cocktail targeting interleukin 10 receptor and transforming growth factor-\u03b2 receptor on dendritic cells potentiates tumour antigen-specific CD8(+) T cell immunity","volume":"181","author":"Ahn","year":"2015","journal-title":"Clin Exp Immunol"},{"key":"B27","doi-asserted-by":"publisher","DOI":"10.1097\/CJI.0b013e31826f79a6","article-title":"Improvement of antitumor effect of intratumoral injection of immature dendritic cells into irradiated tumor by cyclophosphamide in mouse colon cancer model","volume":"35","author":"Son","year":"2012","journal-title":"J Immunother"},{"key":"B28","doi-asserted-by":"publisher","DOI":"10.1097\/CJI.0000000000000054","article-title":"Cyclophosphamide and IL-12-transduced DCs enhance the antitumor activity of tumor antigen-stimulated DCs and reduce tregs and MDSCs number","volume":"37","author":"Rossowska","year":"2014","journal-title":"J Immunother"},{"key":"B29","doi-asserted-by":"publisher","DOI":"10.1158\/1078-0432.CCR-07-0517","article-title":"Low-dose paclitaxel prior to intratumoral dendritic cell vaccine modulates intratumoral cytokine network and lung cancer growth","volume":"13","author":"Zhong","year":"2007","journal-title":"Clin Cancer Res"},{"key":"B30","doi-asserted-by":"publisher","DOI":"10.1002\/prp2.47","article-title":"Hydroxyethyl starch as an effective methotrexate carrier in anticancer therapy","volume":"2","author":"Goszczy\u0144ski","year":"2014","journal-title":"Pharmacol Res Perspect"},{"key":"B31","doi-asserted-by":"publisher","DOI":"10.1021\/acs.biochem.8b00607","article-title":"Internalization of methotrexate conjugates by folate receptor-\u03b1","volume":"57","author":"Nogueira","year":"2018","journal-title":"Biochemistry"},{"key":"B32","doi-asserted-by":"publisher","DOI":"10.3892\/or.2021.7930","article-title":"Immunomodulatory potential of anticancer therapy composed of methotrexate nanoconjugate and dendritic cell\u2212based vaccines in murine colon carcinoma","volume":"45","author":"Szczygie\u0142","year":"2021","journal-title":"Oncol Rep"},{"key":"B33","doi-asserted-by":"publisher","DOI":"10.4049\/jimmunol.0900734","article-title":"Chemotherapeutic agents in noncytotoxic concentrations increase antigen presentation by dendritic cells via an IL-12-dependent mechanism","volume":"183","author":"Shurin","year":"2009","journal-title":"J Immunol"},{"key":"B34","doi-asserted-by":"publisher","first-page":"58","DOI":"10.1186\/1479-5876-7-58","article-title":"Chemomodulation of human dendritic cell function by antineoplastic agents in low noncytotoxic concentrations","volume":"7","author":"Kaneno","year":"2009","journal-title":"J Transl Med"},{"key":"B35","doi-asserted-by":"publisher","first-page":"97","DOI":"10.1007\/s13402-010-0005-5","article-title":"Chemotherapeutic agents in low noncytotoxic concentrations increase immunogenicity of human colon cancer cells","volume":"34","author":"Kaneno","year":"2011","journal-title":"Cell Oncol (Dordr)."},{"key":"B36","doi-asserted-by":"publisher","first-page":"1758835920987056","DOI":"10.1177\/1758835920987056","article-title":"Methotrexate enhances antigen presentation and maturation of tumour antigen-loaded dendritic cells through NLRP3 inflammasome activation: A strategy for dendritic cell-based cancer vaccine","volume":"13","author":"Shi","year":"2021","journal-title":"Ther Adv Med Oncol"},{"key":"B37","doi-asserted-by":"publisher","DOI":"10.1158\/1078-0432.CCR-15-1399","article-title":"Dendritic cell-based immunotherapy: State of the art and beyond","volume":"22","author":"Bol","year":"2016","journal-title":"Clin Cancer Res"},{"key":"B38","first-page":"7","article-title":"Loss of tumorigenicity of murine colon carcinoma MC38\/0 cell line after transduction with a retroviral vector carrying murine IL-12 genes","volume":"50","author":"Pajtasz-Piasecka","year":"2004","journal-title":"Folia Biol (Praha)"},{"key":"B39","doi-asserted-by":"publisher","DOI":"10.3892\/or_00000386","article-title":"Tumour antigen-loaded mouse dendritic cells maturing in the presence of inflammatory cytokines are potent activators of immune response in vitro but not in vivo","volume":"21","author":"Rossowska","year":"2009","journal-title":"Oncol Rep"},{"key":"B40","doi-asserted-by":"publisher","DOI":"10.3389\/fimmu.2017.00713","article-title":"Intratumoral lentivector-mediated TGF-\u03b21 gene downregulation as a potent strategy for enhancing the antitumor effect of therapy composed of cyclophosphamide and dendritic cells","volume":"8","author":"Rossowska","year":"2017","journal-title":"Front Immunol"},{"key":"B41","doi-asserted-by":"publisher","first-page":"7508928","DOI":"10.1155\/2022\/7508928","article-title":"The beneficial effect of IL-12 and IL-18 transduced dendritic cells stimulated with tumor antigens on generation of an antitumor response in a mouse colon carcinoma model","volume":"2022","author":"Mierzejewska","year":"2022","journal-title":"J Immunol Res"},{"key":"B42","doi-asserted-by":"publisher","DOI":"10.3892\/or.17.5.1249","article-title":"Generation of anti-tumor response by JAWS II mouse dendritic cells transduced with murine interleukin 12 genes","volume":"17","author":"Pajtasz-Piasecka","year":"2007","journal-title":"Oncol Rep"},{"key":"B43","doi-asserted-by":"publisher","DOI":"10.1080\/15384101.2018.1471312","article-title":"An engineered novel lentivector specifically transducing dendritic cells and eliciting robust HBV-specific CTL response by upregulating autophagy in T cells","volume":"17","author":"Ma","year":"2018","journal-title":"Cell Cycle"},{"key":"B44","doi-asserted-by":"publisher","DOI":"10.1128\/JVI.00014-10","article-title":"HIV-1 lentiviral vector immunogenicity is mediated by toll-like receptor 3 (TLR3) and TLR7","volume":"84","author":"Breckpot","year":"2010","journal-title":"J Virol"},{"key":"B45","doi-asserted-by":"publisher","DOI":"10.1080\/21645515.2018.1431598","article-title":"Inhibition of IL-10 and TGF-\u03b2 receptors on dendritic cells enhances activation of effector T-cells to kill cholangiocarcinoma cells","volume":"14","author":"Thepmalee","year":"2018","journal-title":"Hum Vaccin Immunother"},{"key":"B46","doi-asserted-by":"publisher","first-page":"901","DOI":"10.1016\/j.ccell.2019.05.005","article-title":"Targeting tumors with IL-10 prevents dendritic cell-mediated CD8+ T cell apoptosis","volume":"35","author":"Qiao","year":"2019","journal-title":"Cancer Cell"},{"key":"B47","doi-asserted-by":"publisher","first-page":"15","DOI":"10.1007\/s10637-020-00972-9","article-title":"Hydroxyethylcellulose as a methotrexate carrier in anticancer therapy","volume":"39","author":"Ciekot","year":"2021","journal-title":"Invest New Drugs"},{"key":"B48","doi-asserted-by":"publisher","first-page":"1748","DOI":"10.3390\/ijms22041748","article-title":"In vitro and In vivo efficacy of a novel glucose-methotrexate conjugate in targeted cancer treatment","volume":"22","author":"Wo\u017aniak","year":"2021","journal-title":"Int J Mol Sci"},{"key":"B49","doi-asserted-by":"publisher","first-page":"75","DOI":"10.1038\/s41392-021-00484-9","article-title":"Targeting tumor-associated macrophages to synergize tumor immunotherapy","volume":"6","author":"Xiang","year":"2021","journal-title":"Signal Transduct Target Ther"},{"key":"B50","doi-asserted-by":"publisher","first-page":"8470","DOI":"10.3390\/ijms22168470","article-title":"Tumor-associated macrophages (TAMs) in colorectal cancer (CRC): From mechanism to therapy and prognosis","volume":"22","author":"Wang","year":"2021","journal-title":"Int J Mol Sci"},{"key":"B51","doi-asserted-by":"publisher","DOI":"10.1158\/0008-5472.CAN-09-4672","article-title":"Different tumor microenvironments contain functionally distinct subsets of macrophages derived from Ly6C(high) monocytes","volume":"70","author":"Movahedi","year":"2010","journal-title":"Cancer Res"},{"key":"B52","doi-asserted-by":"publisher","first-page":"24","DOI":"10.1158\/0008-5472.CAN-13-1196","article-title":"Tumor hypoxia does not drive differentiation of tumor-associated macrophages but rather fine-tunes the M2-like macrophage population","volume":"74","author":"Laoui","year":"2014","journal-title":"Cancer Res"},{"key":"B53","doi-asserted-by":"publisher","DOI":"10.1172\/JCI59643","article-title":"Macrophage plasticity and polarization: In vivo veritas","volume":"122","author":"Sica","year":"2012","journal-title":"J Clin Invest."},{"key":"B54","doi-asserted-by":"publisher","DOI":"10.1158\/0008-5472.CAN-09-3767","article-title":"Immature immunosuppressive CD14+HLA-DR-\/low cells in melanoma patients are Stat3hi and overexpress CD80, CD83, and DC-sign","volume":"70","author":"Poschke","year":"2010","journal-title":"Cancer Res"},{"key":"B55","doi-asserted-by":"publisher","DOI":"10.1016\/j.immuni.2014.10.020","article-title":"Myeloid-derived suppressor activity is mediated by monocytic lineages maintained by continuous inhibition of extrinsic and intrinsic death pathways","volume":"41","author":"Haverkamp","year":"2014","journal-title":"Immunity"},{"key":"B56","doi-asserted-by":"publisher","first-page":"200111","DOI":"10.1098\/rsob.200111","article-title":"Mechanisms of immune suppression by myeloid-derived suppressor cells: The role of interleukin-10 as a key immunoregulatory cytokine","volume":"10","author":"Yaseen","year":"2020","journal-title":"Open Biol"}],"container-title":["Frontiers in Immunology"],"original-title":[],"link":[{"URL":"https:\/\/www.frontiersin.org\/articles\/10.3389\/fimmu.2023.1155377\/full","content-type":"unspecified","content-version":"vor","intended-application":"similarity-checking"}],"deposited":{"date-parts":[[2023,3,23]],"date-time":"2023-03-23T04:47:18Z","timestamp":1679546838000},"score":1,"resource":{"primary":{"URL":"https:\/\/www.frontiersin.org\/articles\/10.3389\/fimmu.2023.1155377\/full"}},"subtitle":[],"short-title":[],"issued":{"date-parts":[[2023,3,23]]},"references-count":56,"alternative-id":["10.3389\/fimmu.2023.1155377"],"URL":"https:\/\/doi.org\/10.3389\/fimmu.2023.1155377","relation":{},"ISSN":["1664-3224"],"issn-type":[{"value":"1664-3224","type":"electronic"}],"subject":[],"published":{"date-parts":[[2023,3,23]]},"article-number":"1155377"}}