18. EVALUATING PROLIFERATION POTENTIAL OF CAR-T CELLS WHEN CO-CULTURED WITH CD19 CANCER CELL LINES IN VITRO
Main Article Content
Abstract
Objective: To evaluate the proliferation ability of CAR-T when co-cultured with CD19+-expressing cell lines in vitro.
Subjects and methods: Transfering gene into K562 cells using the Transposon-transposase vector system combined with electroporation. PBMC cells were eparated and analyzed PBMC by flow cytometry. PBMC were transposed gene using P3 Primary Cells transfection solution, 1 µg SB100X plasmid and 3 µg CD19RCD137/pSB plasmid. Co-culturing CAR-T and CD19+ cells at a ratio of 1:1.
Results: The results showed that on 4-day culture, both CAR-T CD19RCD137 and CAR-T iCasp9-IL15 cells had the ability to proliferate during co-culture with CD19+ cells. The number of CD19RCD137 CAR-T cells tended to decrease, but the number of 4th CAR-T generation still increased during co-cultured with K562 without expressing CD19. On 11-day culture, there were almost no CD19RCD137 CAR-T cells and a significant decrease in the number of iCasp9-IL15 CAR-T cells.
Conclusion: CAR-T cells have the ability to proliferate strongly during co-culture with CD19+ cell lines but have no ability to proliferate during co-culture with CD19- cell lines.
Article Details
Keywords
CAR-T cells, CD19, CD19RCD137 and iCasp9-IL15.
References
[2] Maus M.V, Grupp S.A, Porter D.L, June C.H, The Journal of the American Society of Hematology, Antibody-modified T cells: CARs take the front seat for hematologic malignancies, 2014, 123 (17): 2625-35.
[3] Brentjens R.J, Davila M.L, Riviere I, Park J, Wang X, Cowell L.G et al, CD19-targeted T cells rapidly induce molecular remissions in adults with chemotherapy-refractory acute lymphoblastic leukemia, 2013, 5 (177): 177ra38-ra38.
[4] Grupp S.A, Kalos M, Barrett D, Aplenc R, Porter D.L, Rheingold S.R et al, Chimeric antigen receptor-modified T cells for acute lymphoid leukemia, 2013, 368 (16): 1509-18.
[5] Maude S.L, Frey N, Shaw P.A, Aplenc R, Barrett D.M, Bunin N.J et al, Chimeric antigen receptor T cells for sustained remissions in leukemia, 2014, 371 (16): 1507-17.
[6] Kim G.H, Dang H.N, Phan M.T.T, Kweon S.H, Chun S, Cho D, X-ray as irradiation alternative for K562 feeder cell inactivation in human natural killer cell expansion, 2018, 38 (10): 5767-72.
[7. Hu S.I, Ko M.C, Dai Y.H, Lin H.A, Chen L.C, Huang K.Y et al, Pre-clinical assessment of chimeric antigen receptor t cell therapy targeting CD19+ B cell malignancy, 2020, 8 (9): 584.
[8] Mark D, Terence J.P, James Ryan M, Ana P, Marcus S.N, Alicia G et al, CD19-CAR Cytokine Induced Killer Cells Armored with IL-18 Control Tumor Burden, Prolong Mouse Survival and Result in In Vivo Persistence of CAR-CIK Cells in a Model of B-Cell Acute Lymphoblastic Leukemia, Blood, 2023, 142 (Supplement 1): 6826, 10.1182/blood-2023-178793.
[9] Prativa S, Xin Y, Daniel A, Davide M, Vikram A, David F et al, Mathematical deconvolution of CAR T-cell proliferation and exhaustion from real-time killing assay data. Journal of the Royal Society Interface, 2020, 17 (162): 20190734, 10.1098/RSIF.2019.0734.
[10] Dorota K, Meena M, Katherine A.F, Malek B, Louai L, Peng X et al, Abstract 1362: Metabolic engineering of CAR-T cells overcomes suppressive adenosine signaling and enhances functionality, Cancer Research, 2022, 82 (12_Supplement): 1362, 10.1158/1538-7445.am2022-1362.
[11] Jianghua W, Luo W, Lu T, ZhaoZhao C, Jie Z, Chenggong L et al, Dual Co-Stimulation with CD28 and ICOS Enhances T Cell Activation and Optimizes CAR-T Cell Antitumor Activity, Blood, 2023, 10.1182/blood-2023-181472.