{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,5,21]],"date-time":"2026-05-21T17:17:27Z","timestamp":1779383847116,"version":"3.53.1"},"reference-count":30,"publisher":"MDPI AG","issue":"5","license":[{"start":{"date-parts":[[2026,4,25]],"date-time":"2026-04-25T00:00:00Z","timestamp":1777075200000},"content-version":"vor","delay-in-days":0,"URL":"https:\/\/creativecommons.org\/licenses\/by\/4.0\/"}],"funder":[{"DOI":"10.13039\/501100003196","name":"Italian Ministry of Health","doi-asserted-by":"crossref","award":["T43-AN-01"],"award-info":[{"award-number":["T43-AN-01"]}],"id":[{"id":"10.13039\/501100003196","id-type":"DOI","asserted-by":"crossref"}]}],"content-domain":{"domain":[],"crossmark-restriction":false},"short-container-title":["Pharmaceuticals"],"abstract":"<jats:p>Background\/Objectives: CD19-directed chimeric antigen receptor T (CAR-T) cell therapy induces profound immune remodeling. Nonetheless, biomarkers predicting complete remission (CR) remain poorly defined. We characterized longitudinal cytokine and immune-cell dynamics after CAR-T infusion and identified early immunological features associated with CR. Methods: Longitudinal immune profiling was performed in 18 patients with non-Hodgkin lymphoma, including 14 with relapsed\/refractory diffuse large B-cell lymphoma treated with anti-CD19 CAR-T cells. Peripheral blood was collected at the baseline and days 7, 14, 21, 28, and 60 post-infusion. Multiparameter flow cytometry quantified lymphoid and myeloid subsets and Toll-like receptor (TLR)2 and TLR4 expression. Serum cytokines were measured by multiplex assays. Machine-learning-based feature selection identified variables associated with CR. Results: Two inflammatory waves were observed. The first, at day 7, featured elevated IL-6, IL-10, IFN-\u03b1, IFN-\u03b3, and TNF-\u03b1, accompanied by increased CD4+ T cells, HLA-DRhigh classical monocytes, and non-classical monocytes. The second, at days 21\u201328, showed increased IL-5, IL-6, IL-12, IFN-\u03b3, and GM-CSF, with expansion of CD4+ and CD8+ T cells, regulatory T cells, NK-T cells, and non-classical monocytes. TLR2 expression was significantly upregulated at day 7 on T-cell subsets and on classical and intermediate monocytes. An exploratory feature-selection analysis identified baseline and day-7 TLR2 and TLR4 expression on lymphoid and myeloid cells, early IFN-\u03b3 levels, and monocyte frequencies as variables associated with CR. Conclusions: Together, these data show that anti-CD19 CAR-T therapy induces two coordinated waves of cytokine release and immune-cell activation. Moreover, the findings suggest that early modulation of innate immune features, particularly TLR2 expression, is associated with complete remission, although these biomarker relationships remain exploratory and require validation in larger cohorts.<\/jats:p>","DOI":"10.3390\/ph19050671","type":"journal-article","created":{"date-parts":[[2026,4,29]],"date-time":"2026-04-29T10:34:42Z","timestamp":1777458882000},"page":"671","update-policy":"https:\/\/doi.org\/10.3390\/mdpi_crossmark_policy","source":"Crossref","is-referenced-by-count":0,"title":["Early Immune Signature Features, Including TLR2 and TLR4 Expression, Are Associated with Complete Remission After CD19 CAR-T Cell Therapy"],"prefix":"10.3390","volume":"19","author":[{"ORCID":"https:\/\/orcid.org\/0009-0007-1527-6498","authenticated-orcid":false,"given":"Serena","family":"Di Iasio","sequence":"first","affiliation":[{"name":"Hematopathology Unit, Institute for Stem Cell Biology, Regenerative Medicine and Innovative Therapeutics (ISBReMIT), Fondazione IRCCS \u201cCasa Sollievo della Sofferenza\u201d, Viale Padre Pio, 7, 71013 San Giovanni Rotondo, Italy"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-2107-9104","authenticated-orcid":false,"given":"Chiara","family":"Di Nunzio","sequence":"additional","affiliation":[{"name":"Hematopathology Unit, Institute for Stem Cell Biology, Regenerative Medicine and Innovative Therapeutics (ISBReMIT), Fondazione IRCCS \u201cCasa Sollievo della Sofferenza\u201d, Viale Padre Pio, 7, 71013 San Giovanni Rotondo, Italy"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-6026-6797","authenticated-orcid":false,"given":"Elisabetta","family":"De Santis","sequence":"additional","affiliation":[{"name":"Hematopathology Unit, Institute for Stem Cell Biology, Regenerative Medicine and Innovative Therapeutics (ISBReMIT), Fondazione IRCCS \u201cCasa Sollievo della Sofferenza\u201d, Viale Padre Pio, 7, 71013 San Giovanni Rotondo, Italy"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0009-0002-4292-057X","authenticated-orcid":false,"given":"Concetta","family":"Stella","sequence":"additional","affiliation":[{"name":"Hematopathology Unit, Institute for Stem Cell Biology, Regenerative Medicine and Innovative Therapeutics (ISBReMIT), Fondazione IRCCS \u201cCasa Sollievo della Sofferenza\u201d, Viale Padre Pio, 7, 71013 San Giovanni Rotondo, Italy"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Daniela","family":"Valente","sequence":"additional","affiliation":[{"name":"Department of Hematology and Stem Cell Transplant Unit, Fondazione IRCCS \u201cCasa Sollievo della Sofferenza\u201d, Viale Capuccini, 1, 71013 San Giovanni Rotondo, Italy"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Dalila","family":"Salvatore","sequence":"additional","affiliation":[{"name":"Department of Hematology and Stem Cell Transplant Unit, Fondazione IRCCS \u201cCasa Sollievo della Sofferenza\u201d, Viale Capuccini, 1, 71013 San Giovanni Rotondo, Italy"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Emanuela","family":"Merla","sequence":"additional","affiliation":[{"name":"Department of Hematology and Stem Cell Transplant Unit, Fondazione IRCCS \u201cCasa Sollievo della Sofferenza\u201d, Viale Capuccini, 1, 71013 San Giovanni Rotondo, Italy"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Grazia","family":"Dell\u2019Olio","sequence":"additional","affiliation":[{"name":"Department of Hematology and Stem Cell Transplant Unit, Fondazione IRCCS \u201cCasa Sollievo della Sofferenza\u201d, Viale Capuccini, 1, 71013 San Giovanni Rotondo, Italy"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Costanzo","family":"Padovano","sequence":"additional","affiliation":[{"name":"Hematopathology Unit, Institute for Stem Cell Biology, Regenerative Medicine and Innovative Therapeutics (ISBReMIT), Fondazione IRCCS \u201cCasa Sollievo della Sofferenza\u201d, Viale Padre Pio, 7, 71013 San Giovanni Rotondo, Italy"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Mattia","family":"Colucci","sequence":"additional","affiliation":[{"name":"Hematopathology Unit, Institute for Stem Cell Biology, Regenerative Medicine and Innovative Therapeutics (ISBReMIT), Fondazione IRCCS \u201cCasa Sollievo della Sofferenza\u201d, Viale Padre Pio, 7, 71013 San Giovanni Rotondo, Italy"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0009-0008-2320-2170","authenticated-orcid":false,"given":"Gaja","family":"Bruno","sequence":"additional","affiliation":[{"name":"Hematopathology Unit, Institute for Stem Cell Biology, Regenerative Medicine and Innovative Therapeutics (ISBReMIT), Fondazione IRCCS \u201cCasa Sollievo della Sofferenza\u201d, Viale Padre Pio, 7, 71013 San Giovanni Rotondo, Italy"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-3333-0255","authenticated-orcid":false,"given":"Barbara","family":"Pasculli","sequence":"additional","affiliation":[{"name":"Laboratory of Oncology, Fondazione IRCCS \u201cCasa Sollievo della Sofferenza\u201d, Viale Cappuccini, 1, 71013 San Giovanni Rotondo, Italy"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0001-5308-7072","authenticated-orcid":false,"given":"Mario","family":"Caldarelli","sequence":"additional","affiliation":[{"name":"Department of Translational Medicine and Surgery, Catholic University of Sacred Heart, Largo Agostino Gemelli, 8, 00168 Rome, Italy"},{"name":"Fondazione Policlinico Universitario A. Gemelli, Istituto di Ricerca e Cura a Carattere Scientifico (IRCCS), Largo Agostino Gemelli, 8, 00168 Rome, Italy"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0001-6031-9636","authenticated-orcid":false,"given":"Paola","family":"Parrella","sequence":"additional","affiliation":[{"name":"Laboratory of Oncology, Fondazione IRCCS \u201cCasa Sollievo della Sofferenza\u201d, Viale Cappuccini, 1, 71013 San Giovanni Rotondo, Italy"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Giovanni","family":"Gambassi","sequence":"additional","affiliation":[{"name":"Department of Translational Medicine and Surgery, Catholic University of Sacred Heart, Largo Agostino Gemelli, 8, 00168 Rome, Italy"},{"name":"Fondazione Policlinico Universitario A. Gemelli, Istituto di Ricerca e Cura a Carattere Scientifico (IRCCS), Largo Agostino Gemelli, 8, 00168 Rome, Italy"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0001-5378-8442","authenticated-orcid":false,"given":"Rossella","family":"Cianci","sequence":"additional","affiliation":[{"name":"Department of Translational Medicine and Surgery, Catholic University of Sacred Heart, Largo Agostino Gemelli, 8, 00168 Rome, Italy"},{"name":"Fondazione Policlinico Universitario A. Gemelli, Istituto di Ricerca e Cura a Carattere Scientifico (IRCCS), Largo Agostino Gemelli, 8, 00168 Rome, Italy"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-4967-4204","authenticated-orcid":false,"given":"Angelo","family":"Carella","sequence":"additional","affiliation":[{"name":"Department of Hematology and Stem Cell Transplant Unit, Fondazione IRCCS \u201cCasa Sollievo della Sofferenza\u201d, Viale Capuccini, 1, 71013 San Giovanni Rotondo, Italy"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0003-2607-2080","authenticated-orcid":false,"given":"Vincenzo","family":"Giambra","sequence":"additional","affiliation":[{"name":"Hematopathology Unit, Institute for Stem Cell Biology, Regenerative Medicine and Innovative Therapeutics (ISBReMIT), Fondazione IRCCS \u201cCasa Sollievo della Sofferenza\u201d, Viale Padre Pio, 7, 71013 San Giovanni Rotondo, Italy"}],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"1968","published-online":{"date-parts":[[2026,4,25]]},"reference":[{"key":"ref_1","doi-asserted-by":"crossref","first-page":"116","DOI":"10.1186\/s12967-025-07571-3","article-title":"Meta-analysis of comparing CAR-T and bispecific antibody therapy in relapsed\/refractory indolent B-cell non-Hodgkin\u2019s lymphomas","volume":"24","author":"Zhang","year":"2025","journal-title":"J. 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