All animal procedures were carried out in strict accordance with the recommendations for the proper use and care of laboratory animals (ECC Directive 86/609/EEC). The protocol was approved by the Inter-Institute Bioethics Commission of the Siberian Branch of the Russian Academy of Sciences (SB RAS). The experiments were conducted in the Center for Genetic Resources of Laboratory Animals at the Institute of Cytology and Genetics, SB RAS. Six- to eight-week-old female NOD SCID (CB17-Prkdcscid/NcrCrl) mice with an average weight of 16 to 20 g were used. Tumors were engrafted by inoculating 5 × 106 Raji-FL1 cells in 200 μl of 0.9% saline solution subcutaneously into the left side of mice. Once tumors had reached a palpable volume of at least 50 mm3, mice were randomly assigned to experimental or control groups. Tumor-bearing mice were injected intravenously with 3 × 106 FL1–CAR-Ts, CD19–CAR-Ts, or Myc–CAR-Ts on day 17 after tumor inoculation. Tumor volume was measured with calipers and estimated using the ellipsoidal formula. Animals were euthanized when the volume of the tumor node reached 2 cm3. On the 38th day after tumor inoculation (21st day after CAR-T infusion), animals from each experimental group were used for isolation of blood, spleen, and bone marrow cells. Erythrocytes were lysed with red blood cell lysis buffer (0.15 M NH4Cl, 10 mM NaHCO3, and 0.1 mM EDTA), and cells were stained with antibodies specific for CD3 (for blood samples), CD45RA, and CCR7 and analyzed by a NovoCyte flow cytometer (ACEA Biosciences). The tumors were fixed in 4% neutral buffered formaldehyde for 2 weeks and processed for paraffin sectioning using standard protocols.

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