Investigation of the Impact from IL-2, IL-7, and IL-15 on the Growth and Signaling of Activated CD4+ T Cells

Abstract

While CAR-T therapy is a growing and promising area of cancer research, it is limited by high cost and the difficulty of consistently culturing T-cells to therapeutically relevant concentrations ex-vivo. Cytokines IL-2, IL-7 and IL-15 have been found to stimulate the growth of T cells, however, the optimized combination of these three cytokines for T cell proliferation is unknown. In this study, we designed an integrated experimental and modeling approach to optimize cytokine supplementation for rapid expansion in clinical applications. We assessed the growth data for statistical improvements over no cytokine supplementation and used a systems biology approach to identify genes with the highest magnitude of expression change from control at several time points. Further, we developed a predictive mathematical model to project the growth rate for various cytokine combinations, and investigate genes and reactions regulated by cytokines in activated CD4+ T cells. The most favorable conditions from the T cell growth study and from the predictive model align to include the full range of IL-2 and IL-7 studied, and at lower levels of IL-15 (6 ng/mL or 36 ng/mL). The highest growth rates were observed where either IL-2 or IL-7 was at the highest concentration tested (15 ng/mL for IL-2 and 80 ng/mL for IL-7) while the other was at the lowest (1 ng/mL for IL-2 and 6 ng/mL for IL-7), or where both IL-2 and IL-7 concentrations are moderate-corresponding to condition keys 200, 020, and 110 respectively. This suggests a synergistic interaction of IL-2 and IL-7 with regards to promoting optimal proliferation and survival of the activated CD4+ T cells. Transcriptomic data analysis identified the genes and transcriptional regulators up/down-regulated by each of the cytokines IL-2, IL-7, and IL-15. It was found that the genes with persistent expressing changes were associated with major pathways involved in cell growth and proliferation. In addition to influencing T cell metabolism, the three cytokines were found to regulate specific genes involved in TCR, JAK/STAT, MAPK, AKT and PI3K-AKT signaling. The developed Fuzzy model that can predict the growth rate of activated CD4+ T cells for various combinations of cytokines, along with identified optimal cytokine cocktails and important genes found in transcriptomic data, can pave the way for optimizing activated CD4 T cells by regulating cytokines in the clinical setting.

Improving CAR T-Cell Persistence

“…Other clinical trials (#NCT00586391 and #NCT00709033) and in vitro experiments have supported this observation. For example, IL-7 and IL-15 can induce naïve T-cells to expand into effector memory T subsets, preserve the stem memory T cell (TSCM) phenotype, and increase resistance to apoptosis following repetitive stimulation, while preserving the migration to secondary lymphoid organs [ 95 , 96 , 97 , 98 , 99 , 100 ]. Multiple studies showed that the addition of antioxidants such as N-acetylcysteine to the cell culture during the manufacturing process has also been shown to inhibit effector differentiation and promote the expansion of TSCM cells [ 101 , 102 , 103 ].…”

Cytokine Adjuvants IL-7 and IL-15 Improve Humoral Responses of a SHIV LentiDNA Vaccine in Animal Models

“…We found that our lentiDNA vaccine prototypes did efficiently co-express both vaccine antigens HIV-Env, SIV-Gag p27 (1476 for CAL-SHIV-IN − IRES IL-7 and 823 pg/mL for CAL-SHIV-IN − IRES IL-15) and the cytokines IL-7 and IL-15 (1654 and 125 pg/mL, respectively), which were released in the supernatant of transfected HEK 293T cells ( Figure 1 c,f and Figure 2 A,B). These lower concentrations of cytokines are compatible with physiologic functions but not with cytokine storm-like induction [ 41 ]. To evaluate the function of IL-7 and IL-15 released in the supernatant of transfected cells, we used our lab-adapted PHPC assay on human PBMCs.…”

Inflammatory Response and Oxidative Stress as Mechanism of Reducing Hyperuricemia of Gardenia jasminoides‐Poria cocos with Network Pharmacology

“…IL-7 is a multifunctional cytokine, which can act on a variety of cells. The main function is to promote the development, antiapoptosis, and proliferation of immature B and T cells and to participate in the differentiation and maturation of thymocytes [ 38 , 39 ]. The biological effect of IL-7 is mainly achieved through the binding of IL-7 to the IL-7 receptor (IL-7R).…”