BUSCADOR RECOLECTA

Additional file 1: Supplementary Fig. 1. Study design and Flow-chart., Consejería de Transformación Económica, Industria, Conocimiento y Universidades Instituto de Salud Carlos III Consejería de Salud y Familias, Junta de Andalucía, Peer reviewed

Additional file 2: Supplementary Table 1. Demographical and anthropometrics characteristics of the study population., Consejería de Transformación Económica, Industria, Conocimiento y Universidades Instituto de Salud Carlos III Consejería de Salud y Familias, Junta de Andalucía, Peer reviewed

Additional file 3: Supplementary Table 2. Comorbidities and behavioral factors of residents from the Santa Caridad Nursing Home., Consejería de Transformación Económica, Industria, Conocimiento y Universidades Instituto de Salud Carlos III Consejería de Salud y Familias, Junta de Andalucía, Peer reviewed

Additional file 4: Supplementary Table 3.Thymic activity, Biochemical, Inflammatory and Immunological profiles of the study populations one month after the second dose (T1)., Consejería de Transformación Económica, Industria, Conocimiento y Universidades Instituto de Salud Carlos III Consejería de Salud y Familias, Junta de Andalucía, Peer reviewed

Additional file 5: Supplementary Fig. 2. Levels of Thymosin-α1 in higher vs. lower responders of the older group., Consejería de Transformación Económica, Industria, Conocimiento y Universidades Instituto de Salud Carlos III Consejería de Salud y Familias, Junta de Andalucía, Peer reviewed

Additional file 6: Supplementary Fig. 3. Correlations among Log of anti-S antibody titers at all different study points., Consejería de Transformación Económica, Industria, Conocimiento y Universidades Instituto de Salud Carlos III Consejería de Salud y Familias, Junta de Andalucía, Peer reviewed

Additional file 7: Supplementary Fig. 4. Factors associated with the magnitude of the initial response to the BNT162B2 vaccine at T1 by age-groups., Consejería de Transformación Económica, Industria, Conocimiento y Universidades Instituto de Salud Carlos III Consejería de Salud y Familias, Junta de Andalucía, Peer reviewed

Additional file 8. Detailed methodology., Consejería de Transformación Económica, Industria, Conocimiento y Universidades Instituto de Salud Carlos III Consejería de Salud y Familias, Junta de Andalucía, Peer reviewed

Supplementary Figures: Supplementary Figure 1. Evaluation of the specificity of the detection reagents for the identification of academic CAR-T cells by flow cytometry.-- Supplementary figure 2. Evaluation of the specificity of the detection reagents for the identification of commercial CAR-T cells by flow cytometry.-- Supplementary Figure 3. Commercial CD19 CAR-T cell expansion in the blood of patients with lymphoma.-- Supplementary Figure 4. Comparison of the expansion dynamics of different commercial CD19 CAR-T cell products.-- Supplementary Figure 5. Immunophenotype characterization of non-modified T cells and CAR-T cells at the time of peak expansion in blood of patients with lymphoma.-- Supplementary Figure 6. Immunophenotype characterization of non-modified T cells and CAR-T cells at the time of peak expansion in blood of patients infused with Tisa-cel vs Axi-cel.-- Supplementary Figure 7. Comparison of T cell subsets between leukapheresis and non-modified T and CAR-T cells at the time of peak expansion.-- Supplementary Figure 8. Quantitative determination of the TCR Vβ repertoire of human T lymphocytes by flow cytometry.-- Supplementary Figure 9. Correlation between CAR-T cell expansion in blood of patients with lymphoma and toxicity or response.-- Supplementary Figure 10. Comparison of the toxicity and efficacy of Tisa-cel and Axi-cel products. // Supplementary Table 1. Flow cytometry reagents for the validation of the detection and immune-phenotype characterization of academic and commercial CD19 CAR-T cells, Purpose: CAR-T cell therapy has proven to be a disruptive treatment in the hematology field, however, less than 50% of patients maintain long-term response and early predictors of outcome are still inconsistently defined. Here, we aimed to optimize the detection of CD19 CAR-T cells in blood and to identify phenotypic features as early biomarkers associated with toxicity and outcomes., Experimental design: In this study, monitoring by flow cytometry and digital PCR (dPCR), and immunophenotypic characterization of circulating CAR-T cells from 48 patients treated with Tisa-cel or Axi-cel was performed., Results: Validation of the flow cytometry reagent for the detection of CAR-T cells in blood revealed CD19 protein conjugated with streptavidin as the optimal detection method. Kinetics of CAR-T cell expansion in blood confirmed median day of peak expansion at seven days post-infusion by both flow cytometry and digital PCR. Circulating CAR-T cells showed an activated, proliferative, and exhausted phenotype at the time of peak expansion. Patients with increased expansion showed more severe CRS and ICANs. Immunophenotypic characterization of CAR-T cells at the peak expansion identified the increased expression of co-inhibitory molecules PD1 and LAG3 and reduced levels of the cytotoxicity marker CD107a as predictors of a better long-term disease control., Conclusions: These data show the importance of CAR-T cells in vivo monitoring and identify the expression of PD1LAG3 and CD107a as early biomarkers of long-term disease control after CAR-T cell therapy., This work has been supported by the Instituto de Salud Carlos III (ISCIII), Project RD21/0017/0021, Red Española de Terapias Avanzadas TERAV funded by European Union-NextGenerationEU. “Plan de Recuperación Transformación y Resiliencia” and Consejería de Salud y Familia, Junta de Andalucía PECART-0185-2020-7, PECART-0185-2020 CSYF 2021 – Proyectos Fondos FEDER. Proyectos estratégicos en Investigación en CAR-T. “Monitorización inmune tras tratamiento con células CAR-T: búsqueda de biomarcadores y medición de la actividadmetabólica como predictores de respuesta”., Peer reviewed