INTEGRATED IMMUNOPHENOTYPING REVEALS DISTINCT MULTICOMPARTMENT IMMUNE SIGNATURES ASSOCIATED WITH CYTOKINE RELEASE SYNDROME FOLLOWING ANTI-CD19 CAR-T CELL THERAPY
- 1A.C. Camargo Cancer Center, São Paulo, SP, Brazil
Abstract
Background: Anti-CD19 chimeric antigen receptor (CAR) T-cell therapy has transformed the treatment of relapsed or refractory hematologic malignancies but is frequently limited by cytokine release syndrome (CRS) and immune effector cell-associated neurotoxicity syndrome (ICANS). Although both toxicities arise from immune dysregulation, their underlying multicompartment immune remodeling remains incompletely understood. Objective: To characterize immune signatures associated with CRS and ICANS by integrating T-cell, NK-cell, and monocyte immunophenotypes with systemic cytokine profiles following anti-CD19 CAR-T therapy. Methods: Peripheral blood mononuclear cells (PBMCs) from CAR-T recipients (CAAE: 65750722.3.0000.5432) and healthy controls were analyzed by multiparameter flow cytometry. T cells, NK cells, and monocytes characterized according to activation, differentiation, exhaustion, and cytotoxicity markers. Circulating cytokines were quantified by cytometric bead array (CBA), and longitudinal cellular and soluble immune profiles were integrated to identify multicompartment immune signatures associated with CRS and ICANS. Results: Analysis of the first four patients identified distinct immune trajectories following anti-CD19 CAR-T infusion. Patients who developed CRS and achieved clinical response exhibited preservation of circulating CD4⁺ T cells, greater contraction of CD8⁺ and CD56^dim^ NK-cell compartments, increased frequencies of CD14⁺ monocytes, and lower PD-1 expression on CD8⁺ T cells. In contrast, patients without CRS and without clinical response showed expansion of total CD3⁺ T cells, higher CD38 expression on conventional and CAR⁺ CD4⁺ and CD8⁺ T cells, and increased PD-1 on CD8⁺ and CAR⁺CD8⁺ T cells. HLA-DR expression decreased in both groups, but to a lesser extent in patients with CRS. Memory T-cell compartments also underwent remodeling, with distinct patterns across CD4⁺ and CD8⁺ subsets. Discussion: CRS was not associated with a uniform activation but rather by coordinated multicompartment immune remodeling involving adaptive, innate, and myeloid compartments. Instead, patients with CRS/response showed a CD4 predominance, reduced CD8 and NK-dim cell counts, preserved monocytes, and lower PD-1 levels on CD8 cells; conversely, patients without CRS and without response exhibited marked CD38 induction and an accumulation of CD8⁺ PD-1⁺ cells. Because CRS and clinical response coincided in this preliminary cohort, toxicity-specific immune changes could not be distinguished from response-associated remodeling. Conclusion: CRS following anti-CD19 CAR-T therapy was associated with distinct multicompartment immune remodeling rather than isolated cellular alterations. These preliminary findings support the concept of integrated immune signatures underlying CAR-T toxicity and justify validation in larger cohorts, including responders with and without CRS and patients with ICANS. Larger cohorts, including responding patients with and without CRS, as well as patients with ICANS, are needed to identify specific toxicity signatures.
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BibTeX
@article{gomes2026integrated,
title = {{INTEGRATED IMMUNOPHENOTYPING REVEALS DISTINCT MULTICOMPARTMENT IMMUNE SIGNATURES ASSOCIATED WITH CYTOKINE RELEASE SYNDROME FOLLOWING ANTI-CD19 CAR-T CELL THERAPY}},
author = {Mariana Soares GOMES and Arianne Fagotti GUSMãO and Larissa Campos SOUZA and Ana Luiza Sionti PACHECO and Gabriel ANDRADE DE PAULA and Raquel Garcia DA SILVA and Filipe Freitas DOS SANTOS and Vanessa Anjos BOVOLENTA and Ana Costa CORDEIRO and Bianca Batista CAL and Daniela DELECHIAVE and Sabrina PINHO and Lais SENDA and Marta Maria Moreira LEMOS and Marina Mattos NASCIMENTO and Mariela Pinho CABRAL PICCIN and Jayr FILHO SCHMIDT and Marjorie Vieira BATISTA},
journal = {Hematology, Transfusion and Cell Therapy},
year = {2026},
month = oct,
doi = {10.1016/j.htct.2026.106883},
url = {https://doi.org/10.1016/j.htct.2026.106883},
note = {CC BY},
}
RIS
TY - JOUR
TI - INTEGRATED IMMUNOPHENOTYPING REVEALS DISTINCT MULTICOMPARTMENT IMMUNE SIGNATURES ASSOCIATED WITH CYTOKINE RELEASE SYNDROME FOLLOWING ANTI-CD19 CAR-T CELL THERAPY
AU - Mariana Soares GOMES
AU - Arianne Fagotti GUSMãO
AU - Larissa Campos SOUZA
AU - Ana Luiza Sionti PACHECO
AU - Gabriel ANDRADE DE PAULA
AU - Raquel Garcia DA SILVA
AU - Filipe Freitas DOS SANTOS
AU - Vanessa Anjos BOVOLENTA
AU - Ana Costa CORDEIRO
AU - Bianca Batista CAL
AU - Daniela DELECHIAVE
AU - Sabrina PINHO
AU - Lais SENDA
AU - Marta Maria Moreira LEMOS
AU - Marina Mattos NASCIMENTO
AU - Mariela Pinho CABRAL PICCIN
AU - Jayr FILHO SCHMIDT
AU - Marjorie Vieira BATISTA
PY - 2026
DA - 2026/10/23
JO - Hematology, Transfusion and Cell Therapy
DO - 10.1016/j.htct.2026.106883
UR - https://doi.org/10.1016/j.htct.2026.106883
AB - Background: Anti-CD19 chimeric antigen receptor (CAR) T-cell therapy has transformed the treatment of relapsed or refractory hematologic malignancies but is frequently limited by cytokine release syndrome (CRS) and immune effector cell-associated neurotoxicity syndrome (ICANS). Although both toxicities arise from immune dysregulation, their underlying multicompartment immune remodeling remains incompletely understood. Objective: To characterize immune signatures associated with CRS and ICANS by integrating T-cell, NK-cell, and monocyte immunophenotypes with systemic cytokine profiles following anti-CD19 CAR-T therapy. Methods: Peripheral blood mononuclear cells (PBMCs) from CAR-T recipients (CAAE: 65750722.3.0000.5432) and healthy controls were analyzed by multiparameter flow cytometry. T cells, NK cells, and monocytes characterized according to activation, differentiation, exhaustion, and cytotoxicity markers. Circulating cytokines were quantified by cytometric bead array (CBA), and longitudinal cellular and soluble immune profiles were integrated to identify multicompartment immune signatures associated with CRS and ICANS. Results: Analysis of the first four patients identified distinct immune trajectories following anti-CD19 CAR-T infusion. Patients who developed CRS and achieved clinical response exhibited preservation of circulating CD4⁺ T cells, greater contraction of CD8⁺ and CD56^dim^ NK-cell compartments, increased frequencies of CD14⁺ monocytes, and lower PD-1 expression on CD8⁺ T cells. In contrast, patients without CRS and without clinical response showed expansion of total CD3⁺ T cells, higher CD38 expression on conventional and CAR⁺ CD4⁺ and CD8⁺ T cells, and increased PD-1 on CD8⁺ and CAR⁺CD8⁺ T cells. HLA-DR expression decreased in both groups, but to a lesser extent in patients with CRS. Memory T-cell compartments also underwent remodeling, with distinct patterns across CD4⁺ and CD8⁺ subsets. Discussion: CRS was not associated with a uniform activation but rather by coordinated multicompartment immune remodeling involving adaptive, innate, and myeloid compartments. Instead, patients with CRS/response showed a CD4 predominance, reduced CD8 and NK-dim cell counts, preserved monocytes, and lower PD-1 levels on CD8 cells; conversely, patients without CRS and without response exhibited marked CD38 induction and an accumulation of CD8⁺ PD-1⁺ cells. Because CRS and clinical response coincided in this preliminary cohort, toxicity-specific immune changes could not be distinguished from response-associated remodeling. Conclusion: CRS following anti-CD19 CAR-T therapy was associated with distinct multicompartment immune remodeling rather than isolated cellular alterations. These preliminary findings support the concept of integrated immune signatures underlying CAR-T toxicity and justify validation in larger cohorts, including responders with and without CRS and patients with ICANS. Larger cohorts, including responding patients with and without CRS, as well as patients with ICANS, are needed to identify specific toxicity signatures.
SN - 2531-1379
SN - 2531-1387
ER -