Synergistic effects of IL-4 and TNFα on the induction of B7-H1 in renal cell carcinoma cells inhibiting allogeneic T cell proliferation

Abstract

BackgroundThe importance of B7-H molecules for the T cell/tumor communication and its impact on renal cell carcinoma (RCC) progression and prognosis has been recently described. Cytokine treatment of RCC has earlier been shown to be beneficial in preclinical settings, but its clinical implementation has not proven to be as effective. This might be partially explained by the yet incomplete picture of cellular alterations in tumor cells upon cytokine treatment investigated in detail in this study.MethodsRCC tumor cell lines were treated with different cytokines alone or in combination. The constitutive and/or cytokine-induced expression of cytokine receptors signaling components and B7-H molecules in RCC cells were analysed by qPCR and flow cytometry. A mcherry reporter gene construct containing B7-H1 promoter was cloned and its activity was determined upon transfection in cytokine-stimulated cells. Cytokine pretreated tumor cells were co-cultured with allogeneic CD8+ T cells from healthy donors and T cell proliferation as well as cytokine secretion was determined.ResultsA heterogeneous, but constitutive B7-H1,-H2,-H3 and H4 expression was found on human RCC cell lines. IL-4 and TNFα treatment led to strong synergistic induction of B7-H1 in RCC cells, whereas B7-H2 was only increased by TNFα. In contrast, B7-H3 and B7-H4 expression were not altered by these cytokines. Treatment of RCC cells with TNFα and IL-4 was accompanied by an activation of signaling molecules like NF-κB, IκB and STAT6. The cytokine-mediated up-regulation of B7-H1 was due to transcriptional control as determined by an increased B7-H1 promoter activity in the presence of IL-4 and TNFα. Despite HLA class I and LFA-1 were also increased, the cytokine-mediated up-regulation of B7-H1 was more pronounced and caused an inhibition of allospecifc CD8+ T cell proliferation.ConclusionThus, IL-4 and TNFα, which could be released by immune cells of the tumor microenvironment, are able to control the B7-H1 expression in RCC thereby altering T cell responses. These data are of importance for understanding the complex interplay of tumor cells with immune cells orchestrated by a number of different soluble and membrane bound mediators and for the implementation of check point antibodies directed against B7-H1.

Regulation of PD-L1 expression on murine tumor-associated monocytes and macrophages by locally produced TNF-α

“…Several secreted cytokines or growth factors were considered candidates for the secreted PD-L1 upregulation, based on previous work [25, 26, 30–32], including GM-CSF, MCP-1, IL-10, IL-3, TGF-β, or TNF-α. To help identify the relevant factors, we first screened recombinant cytokines to identify those that most strongly upregulated PD-L1 expression in the bone marrow monocyte system (Fig.…”

“…However, other cytokines have been found to also upregulate PD-L1 expression. IL-4 and TNF-α were found to synergistically upregulate PD-L1 on certain human cancer cell lines [25]. In another study, monocytes were found to upregulate PD-L1 in response to IL-10 and IL-32γ, and TGF-β induced upregulation of PD-L1 expression on B cells [26].…”

Increased antitumor efficacy of ginsenoside Rh 2 via mixed micelles: in vivo and in vitro evaluation

“…Inhibitory effect of Rh2 and Rh2-M on proliferation of A549 cells was detected by MTT assay (Quandt et al., 2014 ). In logarithmic growth phase, A549 cells were digested with trypsin.…”

Programmed cell death ligand‐1: A dynamic immune checkpoint in cancer therapy

“…The B7‐H1 expression is largely controlled by cytokine‐mediated regulation of promoter regions. Quandt et al reported that B7‐H1 expression and alter T‐cell responses are mediated by interleukin‐4 and necrosis factor in renal cell carcinoma (RCC; Quandt, Jasinski‐Bergner, Mueller, Schulze, & Seliger, 2014). The PD‐L1 conjugation with TGF‐β‐STAT3 may contribute to immune dysfunction and act as a potential novel target site for therapeutics (Song, Park, Nam, Choi, & Sung, 2011).…”