Methods of making chimeric antigen receptor-expressing cells
A technology of chimeric antigen receptors and cells, applied in receptors/cell surface antigens/cell surface determinants, biochemical equipment and methods, animal cells, etc., can solve problems such as the impact of therapeutic efficacy in the manufacturing process
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[1241] The present invention is described in further detail by referring to the following experimental examples. These examples are provided for illustrative purposes only and should not be intended to be limiting unless otherwise indicated. Accordingly, the present invention should in no way be construed as limited to the following examples, but rather should be construed to cover any and all variations which become apparent as a result of the teachings provided herein.
example 1
[1242] Example 1: Glycolytic metabolism of CTL019 cells
[1243] Metabolic changes were evaluated in CAR T cell samples from 41 patients with advanced, heavily pretreated, high-risk CLL who received at least one dose of CD19-directed CAR T cells.
[1244] result
[1245] CAR T cells from patients with partial response (PR) or non-response (NR) demonstrated a glycolytic gene signature. CAR-specific stimulation of retroactive cell infusion product samples increased glycolysis and glucose analog uptake ( figure 2 ), which provides evidence that T cells in CLL patients can be metabolically modulated by linking synthetic CARs. CAR-stimulated T cells from PR / NR patients exhibited higher glucose analog uptake relative to CR / PRTD (partial response, transfusion-dependent) patients ( image 3 ), which is consistent with the transcriptional profile of these cells ( Figures 1A to 1D ). Furthermore, blockade of glycolysis using 2-deoxy-D-glucose (2-DG) reduced effector differentiatio...
example 2
[1257] Example 2: STAT3 Pathway Activation in CD19 Expressing Chimeric Antigen Receptor (CAR) T Cells
[1258] CAR T cell samples from patients with advanced, heavily pretreated, high-risk CLL who received at least one dose of CD19-directed CAR T cells were evaluated for STAT3 pathway activation.
[1259] Materials and methods
[1260] patient sample
[1261] Samples were obtained from CLL patients enrolled in the Penn Institutional Review Board (IRB)-approved clinical trial of single-agent CTL019 therapy. All participants provided written informed consent in accordance with the Declaration of Helsinki and the International Conference on Harmonization Guidelines for Good Clinical Practice.
[1262] These studies are registered with ClinicalTrials.gov (ID numbers: NCT01029366, NCT01747486, and NCT02640209).
[1263] Vector production, T cell isolation and CTL019 cell generation
[1264] A lentiviral vector (GeMCRIS 377 0607-793) containing a transgene encoding a CD19-sp...
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