PRE-CLINICAL RESEARCH ON THE APPLICATION OF CAR-T THERAPY IN THE TREATMENT OF ACUTE LYMPHOBLASTIC LEUKEMIA

Abstract

Objectives: To create chimeric antigen receptor (CAR) T cells and evaluate the effect on CD19+ B cells in animal models of acute lymphoblastic leukemia. Methods: An experimental research (In vitro and In vivo) was conducted at the Department of Pathophysiology, Military Medical University, from June 2019 to 2022. Results: Successfully transformed and created CAR-T cells by nucleofection technology; the survival rate of cells after transformation was 48.9 - 60.7%. The expression of CAR molecules on the T cell surface after 21 and 28 days of culture were 66.23% and 97.34%, respectively. CAR-T cells could proliferate vigorously when co-cultured with cancer cells carrying CD19+, and the ability to lyse cancer cells carrying CD19+ after 6 and 24 hours (lysis efficiency was 24.8 - 71.8%, respectively); this ability increased with the proportion of CAR-T cells present in the medium and with the time of co-culture. CAR-T cell block restricted the proliferation of CD19+ cancer cells (through Luciferase activity), prolonged the survival time, and increased the survival rate of the murine model of acute lymphoblastic leukemia. Conclusion: CAR-T cells can grow when co-cultured with CD19 receptor-expressing cells and CD19+ cancer cell lysis. The excellent effect when using generated CAR-T cells to treat a mouse model of acute leukemia was assessed by Luciferase activity associated with CD19+ cancer cells, survival time, and mouse weight.