IL-2 Combined with IL-15 Enhanced the Expression of NKG2D Receptor on Patient Autologous NK Cells to Inhibit Wilms’ Tumor via MAPK Signaling Pathway

Abstract

Objective. The dysfunction of immune surveillance, a hot spot in cancer research, could lead to the occurrence and development in multicancers. However, the potential mechanisms of immunity in Wilms’ tumor (WT) remain unclear on Wilms’ tumor (WT). In this study, we aim to investigate the immune cell in WT and explore the underlying treatment strategy. Method. We quantified stromal and immune scores by using ESTIMATE algorithm based on gene expression matrix of WT patients in TCGA and GEO databases. Different expression genes (DEGs) and functional enrichments were analyzed by R studio and DAVID tools. Flow cytometry, immunofluorescence staining, ELISA assay, and qRT-PCR were used for detecting the NK cells, cytotoxic cytokines (INF-γ, PRF, and GZMB), and NK cell receptor expression, respectively. WT patient autologous NK cells were stimulated by IL-2 and IL-15, and the cytotoxicity of NK cells against WT cell lines was detected by LDH assay. Western blot experiment was used for measuring the MAPK signaling pathway protein maker in NK cells. Results. ESTIMATE indicated that WT tissue had a lower immune score than adjacent kidney tissue. Meanwhile, the low immune score group was associated with poorly outcomes. DEG functional enrichment analysis showed that NK cell-mediated cytotoxicity was significantly different in low and high immune score groups. Although few of proportion of NK cells in WT patients were increased, most of that were significantly lower than normal children. Moreover, the proportion of NK cells and the expression level of INF-γ, PRF, and GZMB in WT tissue were lower than adjacent kidney tissue. Importantly, the NKG2D expression level of NK cells was significantly lower in WT tissue. Furthermore, in vitro, compared with uncultured NK cells, IL-2 and IL-15 could effectively enhance the cytotoxicity of NK cells on killing the WT cell lines. The FACS and WB results showed that the NKG2D and p-PI3K ratio PI3K, MEK1/2, and p-ERK1/2 ratio ERK1/2 were significantly increased in IL-2 and IL15 group compared with uncultured groups. Conclusion. The abnormal NK cell-mediated cytotoxicity may cause the occurrence of WT. Costimulation of WT patients autologous NK cells could effectively enhance the antitumor reaction which involved in activation of NKG2D-mediated MAPK signaling pathway.

Suppression of NK Cell Activation by JAK3 Inhibition: Implication in the Treatment of Autoimmune Diseases

“…JAK/STAT Inhibitor Totally Abolished NK Expansion and Activation. IL-2 and IL-15 could stimulate several downstream signaling pathways for NK proliferation, activation, and immune functions [63][64][65][66][67]. Among these pathways, JAK/STAT pathway is important for NK cell expansion and activation [68].…”

Immune‐check blocking combination multiple cytokines shown curative potential in mice tumor model

“…21,22 It was found that, on the one hand, NK cells can migrate to the tumor microenvironment under the effect of IL-15 and IL-2, improving the cytotoxicity of NK cells and the recognition of tumor cells. [23][24][25] On the other hand, NK cells have immunomodulatory potential and can secrete cytokines and chemokines IFN-γ, TNF-α, CCL5, XCL1, and granulocyte macrophage colony stimulating factor (GM-CSF) to facilitate innate and adaptive cellular antitumor responses. [26][27][28] Attracting immune cells into the tumor microenvironment, as well as the interaction between immune cells, cell relocation, and functional activation greatly affect the anti-tumor immune response.…”

“… 21 , 22 It was found that, on the one hand, NK cells can migrate to the tumor microenvironment under the effect of IL‐15 and IL‐2, improving the cytotoxicity of NK cells and the recognition of tumor cells. 23 , 24 , 25 On the other hand, NK cells have immunomodulatory potential and can secrete cytokines and chemokines IFN‐γ, TNF‐α, CCL5, XCL1, and granulocyte macrophage colony stimulating factor (GM‐CSF) to facilitate innate and adaptive cellular anti‐tumor responses. 26 , 27 , 28 …”