{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,10,1]],"date-time":"2026-10-01T05:57:48Z","timestamp":1790834268260,"version":"4.1.0"},"reference-count":31,"publisher":"Wiley","issue":"6","license":[{"start":{"date-parts":[[2021,4,29]],"date-time":"2021-04-29T00:00:00Z","timestamp":1619654400000},"content-version":"vor","delay-in-days":0,"URL":"http:\/\/onlinelibrary.wiley.com\/termsAndConditions#vor"}],"content-domain":{"domain":["onlinelibrary.wiley.com"],"crossmark-restriction":true},"short-container-title":["Cytometry Part B Clinical"],"published-print":{"date-parts":[[2021,11]]},"abstract":"<jats:title>Abstract<\/jats:title><jats:sec><jats:title>Background<\/jats:title><jats:p>Chimeric antigen receptor\u2010modified T\u2010cells targeting CD19 (CAR\u2010T19) are licensed for treating relapsed\/refractory diffuse large B\u2010cell lymphoma and B\u2010acute lymphoblastic leukemia. Predicting treatment responses and toxicity (e.g., cytokine release syndrome and neurotoxicity) remains a big challenge. CAR\u2010T19 monitoring could increase our understanding of treatment responses and be of relevance to patient management. A robust method for accurate CAR\u2010T19 detection is therefore extremely desirable.<\/jats:p><\/jats:sec><jats:sec><jats:title>Methods<\/jats:title><jats:p>An assay that uses fluorochrome\u2010conjugated human recombinant soluble CD19 was tested against two commercially available CAR\u2010T19 therapies and a CAR\u2010T19 cell line developed in\u2010house. Precision, concordance, and analyte stability were tested using peripheral blood obtained from CAR\u2010T19\u2010treated patients and controls.<\/jats:p><\/jats:sec><jats:sec><jats:title>Results<\/jats:title><jats:p>The assay showed good accuracy, and had a limit of blank for whole blood samples of 0.13%. Reproducibility and inter\u2010operator concordance were satisfactory (CVs &lt;15%). The assay distinguished CAR\u2010T19 from reactive T\u2010cells in cerebrospinal fluid (CSF) from patients with suspected immune effector cell\u2010associated neurotoxicity syndrome (ICANS), and was adapted to study memory T\u2010cell compartments in treated patients.<\/jats:p><\/jats:sec><jats:sec><jats:title>Conclusion<\/jats:title><jats:p>The assay enabled routine monitoring of CAR\u2010T19 in blood and CSF samples. Despite profound cytopenia in many lymphoma patients, results were obtained regularly from only 4\u00a0ml of blood. The assay can be adapted easily to characterize the memory and exhaustion status of CAR\u2010T19 and native T\u2010cells. Importantly, it does not rely on CAR construct specificity; thus, it can be used to detect any CD19\u2010targeted CAR cell. Finally, our validation process can serve as a blueprint for other fluorochrome proteins used to detect CAR cells.<\/jats:p><\/jats:sec>","DOI":"10.1002\/cyto.b.22005","type":"journal-article","created":{"date-parts":[[2021,4,29]],"date-time":"2021-04-29T17:24:06Z","timestamp":1619717046000},"page":"622-631","update-policy":"https:\/\/doi.org\/10.1002\/crossmark_policy","source":"Crossref","is-referenced-by-count":9,"title":["Detection of <scp>CAR\u2010T19<\/scp> cells in peripheral blood and cerebrospinal fluid: An assay applicable to routine diagnostic laboratories"],"prefix":"10.1002","volume":"100","author":[{"ORCID":"https:\/\/orcid.org\/0000-0003-0743-7400","authenticated-orcid":false,"given":"Ulrika","family":"Johansson","sequence":"first","affiliation":[{"name":"SI\u2010HMDS University Hospitals and Weston NHS Foundation Trust  Bristol United Kingdom"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Kathleen","family":"Gallagher","sequence":"additional","affiliation":[{"name":"Cellular Immunotherapy Program Massachusetts General Hospital Cancer Center  Boston Massachusetts USA"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Victoria","family":"Burgoyne","sequence":"additional","affiliation":[{"name":"SI\u2010HMDS University Hospitals and Weston NHS Foundation Trust  Bristol United Kingdom"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Marcela V.","family":"Maus","sequence":"additional","affiliation":[{"name":"Cellular Immunotherapy Program Massachusetts General Hospital Cancer Center  Boston Massachusetts USA"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Keagan S.","family":"Casey","sequence":"additional","affiliation":[{"name":"Cellular Immunotherapy Program Massachusetts General Hospital Cancer Center  Boston Massachusetts USA"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Gabrielle G.","family":"Brini","sequence":"additional","affiliation":[{"name":"Cellular Immunotherapy Program Massachusetts General Hospital Cancer Center  Boston Massachusetts USA"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Matthew J.","family":"Frigault","sequence":"additional","affiliation":[{"name":"Cellular Immunotherapy Program Massachusetts General Hospital Cancer Center  Boston Massachusetts USA"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Jennifer Y.","family":"Yam","sequence":"additional","affiliation":[{"name":"Cellular Immunotherapy Program Massachusetts General Hospital Cancer Center  Boston Massachusetts USA"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Nikesh","family":"Chavda","sequence":"additional","affiliation":[{"name":"Department of Haematology University Hospitals and Weston NHS Foundation Trust  Bristol United Kingdom"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Caroline","family":"Besley","sequence":"additional","affiliation":[{"name":"Department of Haematology University Hospitals and Weston NHS Foundation Trust  Bristol United Kingdom"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Sanne","family":"Lugthart","sequence":"additional","affiliation":[{"name":"Department of Haematology University Hospitals and Weston NHS Foundation Trust  Bristol United Kingdom"},{"name":"Department of Cellular and Molecular Medicine University of Bristol  Bristol United Kingdom"}],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"311","published-online":{"date-parts":[[2021,4,29]]},"reference":[{"key":"e_1_2_6_2_1","first-page":"S49","article-title":"Limit of blank, limit of detection and limit of quantitation","volume":"29","author":"Armbruster D. 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