{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,7,24]],"date-time":"2026-07-24T11:57:31Z","timestamp":1784894251623,"version":"3.55.0"},"reference-count":30,"publisher":"Springer Science and Business Media LLC","issue":"1","license":[{"start":{"date-parts":[[2021,1,20]],"date-time":"2021-01-20T00:00:00Z","timestamp":1611100800000},"content-version":"tdm","delay-in-days":0,"URL":"http:\/\/creativecommons.org\/licenses\/by\/4.0\/"},{"start":{"date-parts":[[2021,1,20]],"date-time":"2021-01-20T00:00:00Z","timestamp":1611100800000},"content-version":"vor","delay-in-days":0,"URL":"http:\/\/creativecommons.org\/licenses\/by\/4.0\/"}],"funder":[{"DOI":"10.13039\/501100004012","name":"Jane ja Aatos Erkon S\u00e4\u00e4ti\u00f6","doi-asserted-by":"publisher","id":[{"id":"10.13039\/501100004012","id-type":"DOI","asserted-by":"publisher"}]},{"name":"State Research Funding of Turku University Hospital","award":["ERVA#13303"],"award-info":[{"award-number":["ERVA#13303"]}]},{"DOI":"10.13039\/501100006306","name":"Sigrid Jus\u00e9liuksen S\u00e4\u00e4ti\u00f6","doi-asserted-by":"publisher","id":[{"id":"10.13039\/501100006306","id-type":"DOI","asserted-by":"publisher"}]},{"DOI":"10.13039\/501100008413","name":"Instrumentariumin Tiedes\u00e4\u00e4ti\u00f6","doi-asserted-by":"publisher","id":[{"id":"10.13039\/501100008413","id-type":"DOI","asserted-by":"publisher"}]},{"DOI":"10.13039\/100008969","name":"Alfred Kordelinin S\u00e4\u00e4ti\u00f6","doi-asserted-by":"publisher","id":[{"id":"10.13039\/100008969","id-type":"DOI","asserted-by":"publisher"}]},{"DOI":"10.13039\/501100007083","name":"Orionin Tutkimuss\u00e4\u00e4ti\u00f6","doi-asserted-by":"publisher","id":[{"id":"10.13039\/501100007083","id-type":"DOI","asserted-by":"publisher"}]},{"name":"Drug Research Doctoral Programme of the University of Turku Graduate School"}],"content-domain":{"domain":["link.springer.com"],"crossmark-restriction":false},"short-container-title":["J Neuroinflammation"],"published-print":{"date-parts":[[2021,12]]},"abstract":"<jats:title>Abstract<\/jats:title><jats:sec><jats:title>Background<\/jats:title><jats:p>Activated macrophages in the experimental model of multiple sclerosis (MS) express folate receptor-\u03b2 (FR-\u03b2), representing a promising target for the treatment of MS. Here, we both evaluated the efficacy of a novel folate-aminopterin construct (EC2319) in a rat focal model of multiple sclerosis (MS) and investigated the utility of<jats:sup>68<\/jats:sup>Ga-labeled 1,4,7-triazacyclononane-1,4,7-triacetic acid-conjugated folate (<jats:sup>68<\/jats:sup>Ga-FOL) for assessing inflammatory lesions. In addition, we investigated whether FR-\u03b2 is expressed in the brain of patients with MS.<\/jats:p><\/jats:sec><jats:sec><jats:title>Methods<\/jats:title><jats:p>Focal delayed-type hypersensitivity experimental autoimmune encephalomyelitis (<jats:italic>f<\/jats:italic>DTH-EAE) was induced in 40 Lewis rats; 20 healthy Lewis rats were used as controls. Rats were divided into six groups according to the duration of disease (control, acute, or chronic) and intervention (vehicle versus EC2319).<jats:sup>68<\/jats:sup>Ga-FOL analyses, histology, and immunofluorescence of the brain were performed to evaluate the efficacy of subcutaneously administered EC2319 on lesion development. Immunofluorescence was used to assess FR-\u03b2 expression in postmortem brain samples from 5 patients with MS and 5 healthy controls.<\/jats:p><\/jats:sec><jats:sec><jats:title>Results<\/jats:title><jats:p>Immunofluorescence and histological analyses revealed significant reductions in FR-\u03b2 expression (<jats:italic>P<\/jats:italic>&lt; 0.05) and lesion size (<jats:italic>P<\/jats:italic>&lt; 0.01), as well as improved inducible nitric oxide synthase\/mannose receptor C type 1 ratios (<jats:italic>P<\/jats:italic>&lt; 0.01) in macrophages and microglia during the chronic but not acute phase of<jats:italic>f<\/jats:italic>DTH-EAE in EC2319-treated rats. The uptake of IV-injected<jats:sup>68<\/jats:sup>Ga-FOL in the brain was low and did not differ between the groups, but the in vitro binding of<jats:sup>68<\/jats:sup>Ga-FOL was significantly lower in EC2319-treated rats (<jats:italic>P<\/jats:italic>&lt; 0.01). FR-\u03b2 positivity was observed in chronically active lesions and in normal-appearing white matter in MS brain samples.<\/jats:p><\/jats:sec><jats:sec><jats:title>Conclusions<\/jats:title><jats:p>EC2319 was well tolerated and attenuated inflammation and lesion development in a rat model of a chronic progressive form of MS. Human MS patients have FR-\u03b2-positive cells in chronically active plaques, which suggests that these results may have translational relevance.<\/jats:p><\/jats:sec>","DOI":"10.1186\/s12974-021-02073-7","type":"journal-article","created":{"date-parts":[[2021,1,20]],"date-time":"2021-01-20T14:03:15Z","timestamp":1611151395000},"update-policy":"https:\/\/doi.org\/10.1007\/springer_crossmark_policy","source":"Crossref","is-referenced-by-count":12,"title":["Efficacy and tolerability of folate-aminopterin therapy in a rat focal model of multiple sclerosis"],"prefix":"10.1186","volume":"18","author":[{"given":"Petri","family":"Elo","sequence":"first","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Xiang-Guo","family":"Li","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Heidi","family":"Liljenb\u00e4ck","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Maria","family":"Gardberg","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Olli","family":"Moisio","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Maxwell","family":"Miner","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Jenni","family":"Virta","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Antti","family":"Saraste","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Madduri","family":"Srinivasarao","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Michael","family":"Pugh","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Philip S.","family":"Low","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Juhani","family":"Knuuti","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Sirpa","family":"Jalkanen","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Laura","family":"Airas","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Yingjuan June","family":"Lu","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-4006-7977","authenticated-orcid":false,"given":"Anne","family":"Roivainen","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"297","published-online":{"date-parts":[[2021,1,20]]},"reference":[{"key":"2073_CR1","doi-asserted-by":"publisher","first-page":"343","DOI":"10.1016\/S1474-4422(06)70410-0","volume":"5","author":"M Rovaris","year":"2006","unstructured":"Rovaris M, Confraveux C, Roberto F, Kappos L, Comi G, Filippi M. Secondary progressive multiple sclerosis: current knowledge and future challenges. Lancet Neurol. 2006;5:343\u201354.","journal-title":"Lancet Neurol"},{"key":"2073_CR2","doi-asserted-by":"publisher","first-page":"221","DOI":"10.1056\/NEJMoa1606468","volume":"376","author":"X Montalban","year":"2017","unstructured":"Montalban X, Hauser SL, Kappos L, et al. Ocrelizumab versus placebo in primary progressive multiple sclerosis. N Engl J Med. 2017;376:221\u201334.","journal-title":"N Engl J Med"},{"key":"2073_CR3","first-page":"205521731667786","volume":"2","author":"EE Longbrake","year":"2016","unstructured":"Longbrake EE, Cross AH, Salter A. Efficacy and tolerability of oral versus injectable disease-modifying therapies for multiple sclerosis in clinical practice. Mult Scler J Exp Transl Clin. 2016;2:2055217316677868.","journal-title":"Mult Scler J Exp Transl Clin"},{"key":"2073_CR4","doi-asserted-by":"publisher","first-page":"64","DOI":"10.1016\/j.clim.2013.10.010","volume":"150","author":"Y Lu","year":"2014","unstructured":"Lu Y, Wollak KN, Cross VA, et al. Folate receptor-targeted aminopterin therapy is highly effective and specific in experimental models of autoimmune uveitis and autoimmune encephalomyelitis. Clin Immunol. 2014;150:64\u201377.","journal-title":"Clin Immunol"},{"key":"2073_CR5","doi-asserted-by":"publisher","first-page":"252","DOI":"10.1186\/s12974-019-1612-3","volume":"16","author":"P Elo","year":"2019","unstructured":"Elo P, Li XG, Liljenb\u00e4ck H, et al. Folate receptor-targeted positron emission tomography of experimental autoimmune encephalomyelitis. J Neuroinflammation. 2019;16:252.","journal-title":"J Neuroinflammation"},{"key":"2073_CR6","unstructured":"Vlahov IR, Leamon CP, You F, et al. Antifolate conjugates for treating inflammation. US patent application. 2018. US20180280528A1."},{"key":"2073_CR7","doi-asserted-by":"publisher","first-page":"183","DOI":"10.1016\/j.rdc.2009.03.009","volume":"35","author":"E Martin-Mola","year":"2009","unstructured":"Martin-Mola E, Balsa A. Infectious complications of biologic agents. Rheum Dis Clin N Am. 2009;35:183\u201399.","journal-title":"Rheum Dis Clin N Am"},{"key":"2073_CR8","doi-asserted-by":"publisher","first-page":"R56","DOI":"10.1186\/ar3304","volume":"13","author":"Y Lu","year":"2011","unstructured":"Lu Y, Stinnette TW, Westrick E, et al. Treatment of experimental adjuvant arthritis with a novel folate receptor-targeted folic acid aminopterin conjugate. Arthritis Res Ther. 2011;13:R56.","journal-title":"Arthritis Res Ther"},{"key":"2073_CR9","doi-asserted-by":"publisher","first-page":"12","DOI":"10.1002\/art.24219","volume":"60","author":"JW Van der Heijden","year":"2009","unstructured":"Van der Heijden JW, Oerlemans R, Dijkmans BA. Folate receptor beta as a potential delivery route for novel folate antagonists to macrophages in the synovial tissue of rheumatoid arthritis patients. Arthritis Rheum. 2009;60:12\u201321.","journal-title":"Arthritis Rheum"},{"key":"2073_CR10","doi-asserted-by":"publisher","first-page":"2724","DOI":"10.3389\/fimmu.2019.02724","volume":"10","author":"J Schniering","year":"2019","unstructured":"Schniering J, Bene\u0161ov\u00e1 M, Brunner M, et al. 18F-AzaFol for detection of folate receptor-\u03b2 positive macrophages in experimental interstitial lung disease - a proof-of-concept study. Front Immunol. 2019;10:2724.","journal-title":"Front Immunol"},{"key":"2073_CR11","doi-asserted-by":"publisher","first-page":"8092781","DOI":"10.1155\/2018\/8092781","volume":"2018","author":"DMSH Chandrupatla","year":"2018","unstructured":"Chandrupatla DMSH, Jansen G, Mantel E, et al. Imaging and methotrexate response monitoring of systemic inflammation in arthritic rats employing the macrophage PET tracer [18F]fluoro-PEG-folate. Contrast Media Mol Imaging. 2018;2018:8092781.","journal-title":"Contrast Media Mol Imaging"},{"key":"2073_CR12","doi-asserted-by":"publisher","first-page":"3103","DOI":"10.1021\/mp4001684","volume":"10","author":"SA Kularatne","year":"2013","unstructured":"Kularatne SA, Belanger MJ, Meng X, et al. Comparative analysis of folate derived PET imaging agents with [18F]-2-fluoro-2-deoxy-d-glucose using a rodent inflammatory paw model. Mol Pharm. 2013;10:3103\u201311.","journal-title":"Mol Pharm"},{"key":"2073_CR13","doi-asserted-by":"publisher","first-page":"1520","DOI":"10.1021\/acs.molpharmaceut.5b00989","volume":"13","author":"Q Chen","year":"2016","unstructured":"Chen Q, Meng X, McQuade P, et al. Synthesis and preclinical evaluation of folate-NOTA-Al18F for PET imaging of folate-receptor-positive tumors. Mol Pharm. 2016;13:1520\u20137.","journal-title":"Mol Pharm"},{"key":"2073_CR14","doi-asserted-by":"publisher","first-page":"9720","DOI":"10.1038\/s41598-018-27618-4","volume":"8","author":"JMU Silvola","year":"2018","unstructured":"Silvola JMU, Li XG, Virta J, et al. Aluminum fluoride-18 labeled folate enables in vivo detection of atherosclerotic plaque inflammation by positron emission tomography. Sci Rep. 2018;8:9720.","journal-title":"Sci Rep"},{"key":"2073_CR15","doi-asserted-by":"crossref","first-page":"1014","DOI":"10.1002\/nbm.1404","volume":"22","author":"S Serres","year":"2009","unstructured":"Serres S, Anthony DC, Jiang Y, et al. Comparison of MRI signatures in pattern I and II multiple sclerosis models. NMR Biomed. 2009;22:1014\u201324.","journal-title":"NMR Biomed"},{"key":"2073_CR16","doi-asserted-by":"publisher","first-page":"967","DOI":"10.1016\/0306-4522(94)00448-E","volume":"64","author":"MK Matyszak","year":"1995","unstructured":"Matyszak MK, Perry VH. Demyelination in the central nervous system following a delayed-type hypersensitivity response to bacillus Calmette-Guerin. Neuroscience. 1995;64:967\u201377.","journal-title":"Neuroscience."},{"key":"2073_CR17","doi-asserted-by":"publisher","first-page":"305","DOI":"10.2967\/jnumed.114.149955","volume":"56","author":"L Airas","year":"2015","unstructured":"Airas L, Dickens AM, Elo P, et al. In vivo PET imaging demonstrates diminished microglial activation after fingolimod treatment in an animal model of multiple sclerosis. J Nucl Med. 2015;56:305\u201310.","journal-title":"J Nucl Med"},{"key":"2073_CR18","doi-asserted-by":"publisher","first-page":"1175","DOI":"10.1093\/brain\/awp070","volume":"132","author":"JM Frischer","year":"2009","unstructured":"Frischer JM, Bramow S, Dal-Bianco A, et al. The relation between inflammation and neurodegeneration in multiple sclerosis brains. Brain. 2009;132:1175\u201389.","journal-title":"Brain."},{"key":"2073_CR19","doi-asserted-by":"publisher","first-page":"438","DOI":"10.1182\/blood-2008-04-150789","volume":"113","author":"W Xia","year":"2009","unstructured":"Xia W, Hilgenbrink AR, Matteson EL, Lockwood MB, Cheng JX, Low PS. A functional folate receptor is induced during macrophage activation and can be used to target drugs to activated macrophages. Blood. 2009;113:438\u201346.","journal-title":"Blood."},{"key":"2073_CR20","doi-asserted-by":"publisher","first-page":"3126","DOI":"10.1002\/art.22082","volume":"54","author":"T Nagai","year":"2006","unstructured":"Nagai T, Tanaka M, Tsuneyoshi Y, Matsushita K, Sunahara N, Matsuda T. In vitro and in vivo efficacy of a recombinant immunotoxin against folate receptor beta on the activation and proliferation of rheumatoid arthritis synovial cells. Arthritis Rheum. 2006;54:3126\u201334.","journal-title":"Arthritis Rheum"},{"key":"2073_CR21","doi-asserted-by":"publisher","first-page":"59","DOI":"10.1016\/j.bbi.2015.11.002","volume":"53","author":"W Kong","year":"2016","unstructured":"Kong W, Hooper KM, Ganea D. The natural dual cyclooxygenase and 5-lipoxygenase inhibitor flavocoxid is protective in EAE through effects on Th1\/Th17 differentiation and macrophage\/microglia activation. Brain Behav Immun. 2016;53:59\u201371.","journal-title":"Brain Behav Immun"},{"key":"2073_CR22","doi-asserted-by":"publisher","DOI":"10.1371\/journal.pone.0054841","volume":"8","author":"C Liu","year":"2013","unstructured":"Liu C, Li Y, Yu J, et al. Targeting the shift from M1 to M2 macrophages in experimental autoimmune encephalomyelitis mice treated with Fasudil. PLoS One. 2013;8:e54841.","journal-title":"PLoS One"},{"key":"2073_CR23","doi-asserted-by":"publisher","first-page":"268","DOI":"10.1016\/j.intimp.2018.12.001","volume":"67","author":"Z Li","year":"2019","unstructured":"Li Z, Liu F, He X, et al. Exosomes derived from mesenchymal stem cells attenuate inflammation and demyelination of the central nervous system in EAE rats by regulating the polarization of microglia. Int Immunopharmacol. 2019;67:268\u201380.","journal-title":"Int Immunopharmacol"},{"key":"2073_CR24","doi-asserted-by":"publisher","first-page":"2","DOI":"10.1177\/1352458510379243","volume":"17","author":"J Mikita","year":"2011","unstructured":"Mikita J, Dubourdieu-Cassagno N, Deloire MS, et al. Altered M1\/M2 activation patterns of monocytes in severe relapsing experimental rat model of multiple sclerosis. Amelioration of clinical status by M2 activated monocyte administration. Mult Scler. 2011;17:2\u201315.","journal-title":"Mult Scler"},{"key":"2073_CR25","doi-asserted-by":"publisher","first-page":"905","DOI":"10.1172\/JCI26836","volume":"116","author":"O Butovsky","year":"2006","unstructured":"Butovsky O, Landa G, Kunis G, Ziv Y, Avidan H, Greenberg N, et al. Induction and blockage of oligodendrogenesis by differently activated microglia in an animal model of multiple sclerosis. J Clin Invest. 2006;116:905\u201315.","journal-title":"J Clin Invest"},{"key":"2073_CR26","doi-asserted-by":"publisher","first-page":"31","DOI":"10.1016\/j.jneuroim.2005.03.008","volume":"164","author":"AA Vilcaes","year":"2005","unstructured":"Vilcaes AA, Degano AL, L\u00f3pez PHH, Nores GA, Roth GA. Antibodies which block anti-myelin basic protein antibodies associated with development of experimental autoimmune encephalomyelitis in Wistar rats. J Neuroimmunol. 2005;164:31\u20136.","journal-title":"J Neuroimmunol"},{"key":"2073_CR27","doi-asserted-by":"publisher","first-page":"517","DOI":"10.1093\/brain\/awh707","volume":"129","author":"L Boven","year":"2006","unstructured":"Boven L, Van Meurs M, Van Zwam M, et al. Myelin-laden macrophages are anti-inflammatory, consistent with foam cells in multiple sclerosis. Brain. 2006;129:517\u201326.","journal-title":"Brain."},{"key":"2073_CR28","doi-asserted-by":"publisher","first-page":"3466","DOI":"10.1182\/blood-2014-11-612721","volume":"125","author":"RC Lynn","year":"2015","unstructured":"Lynn RC, Poussin M, Kalota A, et al. Targeting of folate receptor \u03b2 on acute myeloid leukemia blasts with chimeric antigen receptor\u2013expressing T cells. Blood. 2015;125:3466\u201376.","journal-title":"Blood."},{"key":"2073_CR29","doi-asserted-by":"publisher","first-page":"15","DOI":"10.1016\/B978-0-444-52001-2.00002-9","volume":"122","author":"A Kutzelnigg","year":"2014","unstructured":"Kutzelnigg A, Lassmann H. Pathology of multiple sclerosis and related inflammatory demyelinating diseases. Handb Clin Neurol. 2014;122:15\u201358.","journal-title":"Handb Clin Neurol"},{"key":"2073_CR30","doi-asserted-by":"publisher","first-page":"389","DOI":"10.1053\/j.sult.2006.06.004","volume":"27","author":"A Kamaya","year":"2006","unstructured":"Kamaya A, Weinstein S, Desser TS. Multiple lesions of the spleen: differential diagnosis of cystic and solid lesions. Semin Ultrasound CT MR. 2006;27:389\u2013403.","journal-title":"Semin Ultrasound CT MR"}],"container-title":["Journal of Neuroinflammation"],"original-title":[],"language":"en","link":[{"URL":"http:\/\/link.springer.com\/content\/pdf\/10.1186\/s12974-021-02073-7.pdf","content-type":"application\/pdf","content-version":"vor","intended-application":"text-mining"},{"URL":"http:\/\/link.springer.com\/article\/10.1186\/s12974-021-02073-7\/fulltext.html","content-type":"text\/html","content-version":"vor","intended-application":"text-mining"},{"URL":"http:\/\/link.springer.com\/content\/pdf\/10.1186\/s12974-021-02073-7.pdf","content-type":"application\/pdf","content-version":"vor","intended-application":"similarity-checking"}],"deposited":{"date-parts":[[2024,8,22]],"date-time":"2024-08-22T18:23:45Z","timestamp":1724351025000},"score":1,"resource":{"primary":{"URL":"https:\/\/jneuroinflammation.biomedcentral.com\/articles\/10.1186\/s12974-021-02073-7"}},"subtitle":[],"short-title":[],"issued":{"date-parts":[[2021,1,20]]},"references-count":30,"journal-issue":{"issue":"1","published-print":{"date-parts":[[2021,12]]}},"alternative-id":["2073"],"URL":"https:\/\/doi.org\/10.1186\/s12974-021-02073-7","relation":{},"ISSN":["1742-2094"],"issn-type":[{"value":"1742-2094","type":"electronic"}],"subject":[],"published":{"date-parts":[[2021,1,20]]},"assertion":[{"value":"12 August 2020","order":1,"name":"received","label":"Received","group":{"name":"ArticleHistory","label":"Article History"}},{"value":"5 January 2021","order":2,"name":"accepted","label":"Accepted","group":{"name":"ArticleHistory","label":"Article History"}},{"value":"20 January 2021","order":3,"name":"first_online","label":"First Online","group":{"name":"ArticleHistory","label":"Article History"}},{"value":"All animal experiments were approved by the National Animal Experiment Board of Finland and the Regional State Administrative Agency for Southern Finland (license number: ESAVI\/2979\/2018) and conducted in accordance with the relevant European Union directive. The study of archival human tissues was approved by the Auria Biobank Scientific Steering Committee (decision AB19-2623). According to the Finnish Biobank Act (688\/2012), separate informed consent from individual patients is not needed.","order":1,"name":"Ethics","group":{"name":"EthicsHeading","label":"Ethics approval and consent to participate"}},{"value":"Not applicable.","order":2,"name":"Ethics","group":{"name":"EthicsHeading","label":"Consent for publication"}},{"value":"Michael Pugh and Yingjuan June Lu are full-time employees of Novartis. All other authors declare no potential conflicts of interest.","order":3,"name":"Ethics","group":{"name":"EthicsHeading","label":"Competing interests"}}],"article-number":"30"}}