CAR-T cell therapy has improved survival in patients with relapsed or refractory diffuse large B-cell lymphoma (DLBCL R/R). However, only 65% of patients achieve a complete metabolic response after this treatment. To date, there is no predictive test for therapeutic response after injection of CAR-T cells. Recent studies have shown that the level of trogocytosis by immune cells correlates with the persistence of tumor cells in patients with hematological malignancies. Our main objective is to identify a phenotypic "signature" of trogocytosis predictive of therapeutic response 6 months after injection of CAR-T cells for DLBCL.
For patients
Patient who has given free and informed consent in writing for inclusion in the non-interventional CART-BANK protocol, and orally for the CARTROG protocol,
Patients over 18 years of age at the time of inclusion,
Diagnosis of LDGCB,
Patients who do not meet all the inclusion criteria,
Pregnant or breast-feeding patient,
Patient unable to follow the procedures and/or frequency of visits planned in the trial, for psychological, family, social or geographical reasons,
Patient unable to consent freely to inclusion, under guardianship, curatorship or safeguard of justice.
Parallel
Flow cytometry analysis to determine the level of trogocytosis by effector immune cells in patients
Other interventionFor each patient, a blood sample will be taken at D0 before the CAR-T cells are injected then at D3, D7, D10, D30 after the injection. Flow cytometry analysis will be performed on each sample on the day of collection to determine the level of trogocytosis by effector immune cells (T lymphocytes, NK cells, CAR-T cells) and to define the phenotypic "signature" of trogocytosis.
Flow cytometry analysis to determine the level of trogocytosis by effector immune cells in volunteers
Other interventionFor each healthy volunteer, a single blood sample will be taken at enrollment. Flow cytometry analysis will be performed on each sample on the day of collection to determine the level of trogocytosis of normal lymphocytes and NK cells.
Identification of a phenotypic "signature" of trogocytosis predictive of failure to achieve a complete metabolic response for patients with diffuse large B-cell lymphoma.
Using flow cytometry to determine the level of trogocytosis, the phenotypic "signature" of trogocytosis will be assessed by the AUC : area under the ROC (Receiver operating characteristic) curve; established on circulating CAR-T cells, T lymphocytes and NK cells of patients, and defined as: the percentage of cells aberrantly expressing different tumor antigens normally expressed on lymphoma cells (CD19, CD20, etc.) at the different analysis times and/or the median florescence intensity (MFI) of the expression of these markers at the different analysis times and/or the evolution of these percentages and the MFI between 2 analysis times. Failure to obtain a complete metabolic response will be defined by: the absence of a complete metabolic response on the PET scans of D30, D90 and M6 in the absence of implementation of a new therapeutic line; or by the absence of complete metabolic response on all PET scans carried out before the implementation of a new therapeutic line.
Identification of a phenotypic "signature" of trogocytosis predictive of the occurrence of grade II or more immunological adverse events
Using flow cytometry analysis, the phenotypic "signature" will be assessed by the AUC used to predict the occurrence of immunological side effects (Cytokine Release Syndrome and immune effector cell-associated neurotoxicity syndrome) of grade II or + according to the American Society for Transplantation and Cellular Therapy ASTCT2 criteria. It will be established on circulating CAR-T, T lymphocytes and NK cells of patients, and defined as : the percentage of cells aberrantly expressing different tumor antigens normally expressed on lymphoma cells at the different analysis times,and/or the MFI of the expression of these markers at the different analysis times and/or the evolution of these percentages and the MFI between 2 analysis times.The occurrence of immunological side effects between D0 and D60 will be assessed by clinical and, if necessary, paraclinical examinations (lumbar puncture, brain MRI, etc.). The grade will be evaluated according to the ASTCT2 criteria.
Identification of a phenotypic "signature" of trogocytosis predictive of the occurrence of serious hematological side effects.
Flow cytometry analysis will be performed to determine the level of trogocytosis by effector cells. The phenotypic "signature" of trogocytosis will be assessed by the AUC used to predict the occurrence of serious hematological side effects (grade III or + cytopenias according to Common Terminology Criteria for Adverse Events CTCAE 5.03 between D30 and M6 after injection of anti-CD19 CAR-T cells .It will be established on circulating CAR-T, T lymphocytes and NK cells of patients, and defined as :the percentage of cells aberrantly expressing different tumor antigens normally expressed on lymphoma cells at the different analysis times, and/or the MFI of the expression of these markers at the different analysis times and/or the evolution of these percentages and the MFI between 2 analysis times. The occurrence of hematological side effects between D30 and M6 will be assessed by blood counts. The grade will be evaluated according to the CTCAE v5.03 criteria.
Determination of the trogocytosis level of normal lymphocytes and NK cells in healthy subjects
Flow cytometry analysis will be performed on each sample on the day of collection (at enrollment), to determine the level of trogocytosis which will be evaluated by simple analysis of the trogocytosis phenotype of normal lymphocytes and NK cells in healthy subjects. This result will be considered as a negative control to determine the phenotypic "signature" of trogocytosis of the primary and secondary outcomes above.
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