Additional file 5 of A novel CAR-T cell product targeting CD74 is an effective therapeutic approach in preclinical mantle cell lymphoma models

Additional file 5: Figure S5. No significant depletion of circulating immune cells pre/post peak detection of 42105-74bbz CAR-T cells in a humanized mouse model. A Absolute cell numbers of B cells, monocytes, G-MDSC, M-MDSC and NK cells in humanized NSG mice on Day 3, 11 and 23 post UTT or 74bbz CAR-T engraftment. All human cells were identified by human CD45+. B: CD33−CD19+; Monocyte: LIN−CD45+CD11b+CD33+CD14+; G-MDSC: LIN−CD11b+CD33+CD14−HLA-DR−; M-MDSC: LIN−CD11b+CD33+CD14+HLA-DR−; NK: CD33−CD3−CD56+. Mice received either UTT cells (red, n = 5) and 42105-74bbz (blue, n = 7). Bars show the median cell number. B CD74 expression (blue) of B cells, monocytes, G-MDSC, M-MDSC and NK cells on Day 18 compared to isotype control (red). Data are from one mouse from the 42105-74bbz CAR-T cell treatment group

Additional file 3 of A novel CAR-T cell product targeting CD74 is an effective therapeutic approach in preclinical mantle cell lymphoma models

Additional file 3: Figure S3. Expression of CD74 after activation on T cell and B cell. T cells and B cells isolated from PBMCs of 3 healthy blood donors were either untreated (red) or activated (blue) by CD3/CD28 soluble antibodies and IL-2 for T cells, and LPS (10 ng/mL)/ anti-IgM (10 µg/mL) for B cells. One representative of 3 healthy blood donors was shown

Additional file 4 of A novel CAR-T cell product targeting CD74 is an effective therapeutic approach in preclinical mantle cell lymphoma models

Additional file 4: Figure S4. Expression of CD74 on human CD34+ stem cells. Human umbilical cord blood mononuclear cells were stained with anti-CD34 and CD74 antibodies and analyzed by flow cytometry. One representative of 3 healthy cord blood donors was shown

Additional file 1 of A novel CAR-T cell product targeting CD74 is an effective therapeutic approach in preclinical mantle cell lymphoma models

Additional file 1: Figure S1. In silico modeling of CD74-anti-CD74 scFV interaction. A Best generated models of CD74-anti-CD74 scFV interaction shown by the lowest HADDOCK score as a function of RMSD. The blue cluster was picked for further in silico mutagenesis. B Visualization of CD74-anti-CD74 scFV interaction. Red: CD74 trimer; Blue: anti-CD74 scFV

Additional file 2 of A novel CAR-T cell product targeting CD74 is an effective therapeutic approach in preclinical mantle cell lymphoma models

Additional file 2: Figure S2. Creation of the 74bbz mutant clones. A GFP+ cells of the 74bbz mutants and parent CAR expressing Jurkat cells were sorted at the same intensity by flow cytometry. B An immunoblot of CD3ζ to show the expressing of parent, 543, 5311, 42105-74bbz clones. Endogenous CD3ζ was detected at 15 kDa while the chimeric CD3ζ on CAR was detected at 55 kDa. C CD74-ECD-Fc fusion protein was stained by Coomassie blue staining

Additional file 6 of A novel CAR-T cell product targeting CD74 is an effective therapeutic approach in preclinical mantle cell lymphoma models

Additional file 6: Figure S6. No significant change in the body weights of the 74bbz CAR-T cells-treated mice was observed. The body weights of the mice treated UTT, 19bbz CAR-T and 74bbz CAR-T cells (n = 10 per group) were monitored until the mice reached ERC and plotted over time