IL9 cell screening model

IL-9 was originally considered a T cell growth factor. The human and mouse IL-9 genes are located on the long arms of chromosomes 5 and 13 respectively, and the human IL-9 gene is located in the Th2 cytokine gene cluster. Medium. IL-9 protein is a 14kD glycosylated protein with 144 amino acids and an 18 amino acid signal peptide. It was originally classified as a cytokine produced by Th2 cells. In recent years, a new IL-9-producing CD4+Th9 cell subset has been discovered and defined, which is differentiated from naive CD4+T cells by transforming growth factor (TGF)-β and IL-4. Is the main cell subpopulation that produces high levels of IL-9. However, IL-9 is not specifically produced by Th9 cells; it can also be produced by innate immune cells such as mast cells and type 2 innate lymphocytes, as well as Th2 cells, regulatory T (Treg) cells, and natural killer (NK) T cells. Produced in low amounts; in addition, some studies have shown that under Th9 polarization conditions, naive CD8+T cells can also differentiate into IL-9-producing CD8+T (Tc9) cells.

Introduction to IL-9

IL-9 transmits signals through its IL-9 receptor complex, which includes the IL-9Rα chain and the common γ chain (γc), and is expressed on B cells and T cells, epithelial cells, and mast cells. In summary, IL-9 is a pleiotropic cytokine that can serve as a positive or negative regulator of a variety of cellular immune responses. Previous studies have shown that IL-9 plays an important role in a variety of physiological and pathological processes, including allergic inflammatory processes, immunity to parasites, and autoimmune diseases. IL-9 mainly promotes the production of IgE by B cells. , epithelial cells produce chemokines and mucus secretion and mast cell proliferation, which play a role in allergic diseases. Therefore, IL-9 and its receptor-related signaling pathways were initially used as targets for the development of immune disease drugs. The anti-IL-9 monoclonal antibody Enokizumab (MEDI528) is the first related signaling pathway drug developed to treat severe asthma. As an IL-9 antagonist, MEDI-528 can inhibit a series of asthma in antigen-exposed mice. Although subsequent clinical trials were not successful, further studies revealed that certain subgroups may benefit from anti-IL-9 treatment if patients with different asthma types are grouped.

In addition to being closely related to immune diseases, the IL-9 signaling pathway has also been found to be closely related to tumor immunity in recent years. It can regulate tumor immunity through various ways to exert anti-tumor functions. First, IL-9 can directly produce anti-tumor effects on a variety of solid tumor cells. The study by Fang et al. reported for the first time the direct anti-melanoma potential of IL-9 in human melanoma cell lines. In several melanoma cell lines, IL-9 strongly inhibited HTB-72 and SK-Mel-5 in vitro. Melanoma growth and induces apoptosis in HTB-72 cells. This growth inhibition and apoptosis are related to the upregulation of the anti-proliferative molecule p21 and the pro-apoptotic molecule TNF-related apoptosis-inducing ligand (TRAIL); while Miao et al. Studies have found that specific Th9 cells may induce apoptosis of cancer squamous cells (SqC) because SqC cells express IL-9R. These studies indicate that, in addition to enhancing anti-tumor immunity, IL-9 can also directly induce apoptosis of IL-9R+ tumor cells. Secondly, IL-9 can exert anti-tumor effects by inducing innate immune responses. Studies have shown that the entry of mast cells into solid tumor sites is related to improved cancer prognosis. In addition to being a T cell factor, IL-9 is also considered a A mast cell growth factor that promotes mast cell activation and function. Purwar et al. found that treatment with recombinant IL-9 failed to inhibit tumor growth in mast cell-deficient mice, suggesting that the presence of mast cells is critical for recombinant IL-9 to exert its antitumor activity, while Abdul-Wahid et al. further found that, IL-9 secreted by Th9 cells blocks cancer cell migration and is accompanied by mast cell activation, whereas inhibition of mast cell degranulation abrogates the antitumor effects mediated by vaccine-induced IL-9 production. In summary, IL-9 can exert anti-tumor effects by targeting the activation and function of mast cells, which acquire direct cytotoxic activity and induce immune cell recruitment and activation against tumor cells. In addition, in addition to inducing innate immune responses, IL-9 can also induce adaptive anti-tumor immune responses. Studies have found that the anti-tumor effects of IL-9 are closely related to tumor-reactive leukocyte subpopulations (including CD4+ T cells) in tumor tissues. Consistent with the significant increase in infiltration of CD8+ T cells), in IL-9-deficient mice, the activation of CD8+ T cells was inhibited and the infiltration of leukocytes was reduced, indicating that Th9/IL-9 can also produce adaptive immune response and exert anti-tumor effects.

R&D status

Some current clinical studies have shown that the Th9/IL-9 signaling pathway is clinically beneficial to the effect of tumor immunotherapy. According to statistics, about 40% of patients with advanced melanoma will respond to Nivolumab (anti-PD-1 antibody). Increases in physiological Th9 cell counts during nivolumab treatment are positively correlated with improved clinical response in patients with metastatic melanoma Nonomura et al. demonstrate that Th9 cells favor melanoma-specific CD8 in patients treated with nivolumab + T cell-mediated anti-tumor function. Subsequently, You et al. further found that in the peripheral blood and circulating CD4+ T cells of breast cancer patients, Th9 cells expressing IL-9 and IL-9 were significantly increased, and through enhanced CD8+ T cell-mediated cytotoxicity exerts anti-tumor effects; in addition, Forget et al. also found that IL-9 is a potential pre-treatment predictive biomarker that can be used to predict the efficacy of adoptive TIL therapy in melanoma patients. evaluation, and high IL-9 production was closely associated with optimal efficacy of adoptive Liandong therapy. In summary, the development of IL-9-based cancer immunotherapy has potential therapeutic significance in clinical practice.

IL9 cell screening model

In view of this potential target of immune diseases and tumor immunity, we have developed a cell-level reporter gene detection model, which can be used to screen and test the activity of drugs related to the IL-9 signaling pathway.

hIL9 Effector Reporter Cell RQP74207

Figure 1. Dose Response of Recombinant Human lL9 in hlL9 Effector Reporter Cells (C18).

Figure 2. Inhibition of Human lL9-induced Reporter Activity by hlL9 Neutralization Ab in hIL9 Effector Reporter Cells (C18)