Compositions and methods for treating t cell exhaustion
A cell, immune cell technology, applied in biochemical equipment and methods, chemical instruments and methods, drug combinations, etc.
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[0186] The present technology is further demonstrated by the following examples, which should not be construed as limiting in any way. The examples herein are compositions and systems provided to demonstrate the advantages of the techniques of the invention and to further assist those of ordinary skill in the art in preparing or using the techniques of the invention. The examples should in no way be construed as limiting the technical scope of the present invention defined by the appended claims. An example may comprise or incorporate any variation, aspect or embodiment of the inventive techniques described above. The variations, aspects or embodiments described above may also each further comprise or incorporate variations of any or all other variations, aspects or embodiments of the present technology.
example 1
[0187] Example 1: Isolated immune cells express reduced levels of immune checkpoint receptors PD-1 and CTLA-4 after treatment with TAT-MYC
[0188] A whole blood sample is isolated from a donor subject and mixed with a blood anticoagulant, ethylenediaminetetraacetic acid (EDTA). At 37°C, 5% CO 2 After incubating the cells for at least 24 hours at 10°C, whole blood samples were then separated into Peripheral blood mononuclear cells (PBMC; immune cells) including lymphocytes (eg, T cells, B cells, NK cells) and monocytes, as well as waste products (ie, red blood cells, platelets, plasma, etc.) . During the cell isolation process, PBMCs were washed twice with 2.5% (w / v) HSA (human serum albumin) saline solution. After the washing step, the immune cells were resuspended at a concentration of 1 x 10 6 cells / ml in 2.5% (w / v) HSA solution to provide a cell suspension.
[0189] After the cell isolation process, with DPBS (negative control) or TAT-MYC fusion protein (for 10 6 cel...
example 2
[0192] Example 2: Isolated Immune Cells Express Reduced Expression of Dysfunctional Immune Cell Markers Following Treatment with TAT-MYC
[0193] Peripheral blood mononuclear cells (PBMC) were prepared as described above. After the cell isolation process, with DPBS (negative control) or TAT-MYC fusion protein (for 10 6 cells, 25 μg / mL) and incubate the sample for 1 hour at room temperature. Then, the treated immune cells (called TBX-3400) were washed again on SEPAX-100, and excess TAT-MYC was washed from the cells with 2.5% (w / v) HSA solution. After the final wash step, resuspend TBX-3400 at a concentration of 1 x 10 6 cells / ml in 2.5% (w / v) HSA solution.
[0194] 24-well plates were coated with anti-CD3e antibody solution (500 μL, 5 μg / mL; BD Biosciences) in sterile DPBS. For control wells, only 500 μL of DPBS was added. Plates were incubated overnight at 4°C before the solution was removed. Each well was then washed twice with 2 mL of sterile DPBS. Then, the cells wer...
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