{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,7,15]],"date-time":"2026-07-15T16:40:41Z","timestamp":1784133641307,"version":"3.55.0"},"reference-count":55,"publisher":"Wiley","issue":"2","license":[{"start":{"date-parts":[[2023,9,27]],"date-time":"2023-09-27T00:00:00Z","timestamp":1695772800000},"content-version":"vor","delay-in-days":0,"URL":"http:\/\/creativecommons.org\/licenses\/by-nc\/4.0\/"}],"funder":[{"DOI":"10.13039\/501100005010","name":"Associazione Italiana per la Ricerca sul Cancro","doi-asserted-by":"publisher","award":["22791"],"award-info":[{"award-number":["22791"]}],"id":[{"id":"10.13039\/501100005010","id-type":"DOI","asserted-by":"publisher"}]}],"content-domain":{"domain":["onlinelibrary.wiley.com"],"crossmark-restriction":true},"short-container-title":["Cytometry Pt A"],"published-print":{"date-parts":[[2024,2]]},"abstract":"<jats:title>Abstract<\/jats:title><jats:p>CD19\u2010targeted chimeric antigen receptor T (CAR\u2010T) cell therapy has shown unprecedented results in patients with B cell relapsed\/refractory acute lymphoblastic leukemia (R\/R\u2010ALL) and B cell non\u2010Hodgkin lymphomas where no other curative options are available. In vivo monitoring of CAR\u2010T cell kinetics is fundamental to understand the correlation between CAR\u2010T cells expansion and persistence with treatment response and toxicity development. The aim of this study was to define a robust, sensitive, and universal method for CAR\u2010T cell detection using flow cytometry. We set up and compared with each other three assays for CD19 CAR\u2010T cell detection, all based on commercially available reagents. All methods used a recombinant human CD19 protein fragment recognized by the single\u2010chain variable fragment of the CAR construct. The two indirect staining assays (CD19his\u2009+\u2009APC\u2010conjugated antihistidine antibody and CD19bio\u2009+\u2009APC\u2010conjugated antibiotin antibody) showed better sensitivity and specificity compared with the direct staining with CD19\u2010FITC, and CD19his had a better cost\u2010effective profile. We validated CAR detection with CD19his with parallel quantitative real\u2010time polymerase chain reaction data and we could demonstrate a strong positive correlation. We also showed that CD19his staining can be easily included in a multicolor flow cytometry panel to achieve additional information about the cell phenotype of CAR\u2010T cell positive subpopulations. Finally, this method can be used for different anti\u2010CD19 CAR\u2010T cell products and for different sample sources. These data demonstrate that detection of CAR\u2010T cells by CD19his flow cytometry staining is a reliable, robust, and broadly applicable tool for in vivo monitoring of CAR\u2010T cells.<\/jats:p>","DOI":"10.1002\/cyto.a.24796","type":"journal-article","created":{"date-parts":[[2023,9,14]],"date-time":"2023-09-14T17:44:31Z","timestamp":1694713471000},"page":"112-123","update-policy":"https:\/\/doi.org\/10.1002\/crossmark_policy","source":"Crossref","is-referenced-by-count":8,"title":["Optimization and validation of in vivo flow cytometry <scp>chimeric antigen receptor T<\/scp> cell detection method using <scp>CD19his<\/scp> indirect staining"],"prefix":"10.1002","volume":"105","author":[{"given":"Silvia","family":"Zaninelli","sequence":"first","affiliation":[{"name":"Division of Hematology, Center of Cellular Therapy \u201cG. Lanzani\u201d ASST Papa Giovanni XXIII Hospital  Bergamo Italy"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Cristian","family":"Meli","sequence":"additional","affiliation":[{"name":"Hematology and Bone Marrow Transplant Unit ASST Papa Giovanni XXIII Hospital  Bergamo Italy"},{"name":"Master of Science Programme in Biology Applied to Research in Biomedicine, Facolt\u00e0 di Scienze e Tecnologie Universit\u00e0 degli Studi di Milano  Milan Italy"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Gianmaria","family":"Borleri","sequence":"additional","affiliation":[{"name":"Hematology and Bone Marrow Transplant Unit ASST Papa Giovanni XXIII Hospital  Bergamo Italy"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Michele","family":"Quaroni","sequence":"additional","affiliation":[{"name":"Laboratory of Cell and Gene Therapy Stefano Verri Fondazione IRCCS San Gerardo dei Tintori  Monza Italy"},{"name":"M. Tettamanti Center Fondazione IRCCS San Gerardo dei Tintori  Monza Italy"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Chiara","family":"Pavoni","sequence":"additional","affiliation":[{"name":"Hematology and Bone Marrow Transplant Unit ASST Papa Giovanni XXIII Hospital  Bergamo Italy"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Giuseppe","family":"Gaipa","sequence":"additional","affiliation":[{"name":"Laboratory of Cell and Gene Therapy Stefano Verri Fondazione IRCCS San Gerardo dei Tintori  Monza Italy"},{"name":"M. Tettamanti Center Fondazione IRCCS San Gerardo dei Tintori  Monza Italy"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Andrea","family":"Biondi","sequence":"additional","affiliation":[{"name":"Laboratory of Cell and Gene Therapy Stefano Verri Fondazione IRCCS San Gerardo dei Tintori  Monza Italy"},{"name":"M. Tettamanti Center Fondazione IRCCS San Gerardo dei Tintori  Monza Italy"},{"name":"Department of Pediatrics University of Milano \u2013 Bicocca  Monza Italy"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Martino","family":"Introna","sequence":"additional","affiliation":[{"name":"Division of Hematology, Center of Cellular Therapy \u201cG. Lanzani\u201d ASST Papa Giovanni XXIII Hospital  Bergamo Italy"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Jos\u00e9e","family":"Golay","sequence":"additional","affiliation":[{"name":"Division of Hematology, Center of Cellular Therapy \u201cG. 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