Tethered interleukin-15 and interleukin-21
A cell, host cell technology, applied in the direction of interleukins, receptors/cell surface antigens/cell surface determinants, cytokines/lymphokines/interferons, etc., can solve T cell persistence, survival and anti-tumor activity lowering etc.
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Embodiment 1
[0191] This example demonstrates that cells transduced with TeIL-15 Lr1 exhibit superior proliferation in the absence of exogenous IL-2 compared to other IL-15 constructs in Table 2.
[0192] PBMCs were cultured in AIM V medium supplemented with IL-2 (300 IU / mL) and anti-CD3 antibody (OKT3, 50 ng / mL) for 2 days. Cells were then transduced with lentiviruses containing the TeIL-15 Lr1, TeIL-15 Lr2, IL-15 RA, TeIL-15 Lr6, or IL-15S constructs in Table 2. The same number of cells transduced with each IL-15 construct was plated on day 0 when IL-2 was withdrawn from the medium at day 7 post-transduction ( figure 2 ). through the figure 2 Proliferation of transduced cells was assessed using the trypan blue assay to assess the total number of viable cells at each of the different time points indicated. Untransduced cells were used as negative controls. Untransduced cells cultured in the presence of exogenous IL-15 were used as positive controls.
[0193] The results are shown i...
Embodiment 2
[0195] This example demonstrates that cells transduced with the TeIL-15 Lr1 construct exhibit the highest IL- 15 expression levels.
[0196] PBMC were transduced with lentiviruses containing the TeIL-15 Lr1, TeIL-15 Lr2, IL-15 RA or IL-15S constructs in Table 2 as described in Example 1. The presence of IL-15 expressed on the cell membrane was analyzed by flow cytometry. Cells transduced with the TeIL-15Lr1 construct exhibited Highest IL-15 expression level (62.3%).
Embodiment 3
[0198] This example demonstrates that 293T cells transduced with the TeIL-21 Lr6, TeIL-21 Lr1 or TeIL-21 Lr2 constructs in Table 4 express IL-21 on the cell surface.
[0199] Cells (293T) were transduced with lentiviruses containing the TeIL-21 Lr7, TeIL-21 Lr6, TeIL-21 Lr1 or TeIL-21 Lr2 constructs in Table 4. Unstained cells and stained but not transduced cells were used as negative controls.
[0200] TeIL-21 expression was analyzed by flow cytometry. The results are shown in Table 7. As shown in Table 7, more than 30% of 293T cells transduced with TeIL-21 Lr6, TeIL-21 Lr1 or TeIL-21 Lr2 expressed IL-21.
[0201] Table 7
[0202]
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