IL-4 inhibits the TNF-alpha induced proliferation of renal cell carcinoma (RCC) and cooperates with TNF-alpha to induce apoptotic and cytokine responses by RCC: implications for antitumor immune responses - PubMed (original) (raw)

IL-4 inhibits the TNF-alpha induced proliferation of renal cell carcinoma (RCC) and cooperates with TNF-alpha to induce apoptotic and cytokine responses by RCC: implications for antitumor immune responses

Claudia Falkensammer et al. Cancer Immunol Immunother. 2006 Oct.

Abstract

Objective: While previous reports clearly demonstrated antiproliferative effects of IL-4 on renal cell carcinoma (RCC) in vitro, the administration of IL-4 to patients with metastatic RCC in clinical trials could not recapitulate the promising preclinical results. In the present study we wanted to examine the context of IL-4 action and to establish conditions of enhanced IL-4 efficacy.

Methods: Primary and permanent human RCC cells were cultured in either serum-supplemented or chemically defined, serum-free culture medium in the presence or absence of cytokines. Cell proliferation was assessed as [(3)H]-thymidine incorporation. Cell apoptosis was measured using the fluorescent DNA intercalator 7-aminoactinomycin D and flow cytometry. In addition, culture media conditioned by RCC were subjected to cytokine antibody array and cytokine multiplex analysis.

Results: Our results indicate that the previously reported antiproliferative effects of IL-4 are serum-dependent. Under serum-free conditions, IL-4 failed to exhibit growth-inhibitory effects or was even growth-stimulatory. In a chemically defined, serum-free medium (AIM-V), however, IL-4 inhibited the TNF-alpha induced proliferation of RCC. IL-4 and TNF-alpha synergistically induced apoptosis of RCC as well as a complex cytokine response by RCC, which included the synergistic upregulation of RANTES and MCP-1.

Conclusions: IL-4 alone has little effect on the spontaneous proliferation of RCC but can prevent the enhancement of proliferation induced by growth promoters like FBS and TNF-alpha. The concomitant growth inhibitory, apoptosis-inducing, and cytokine-enhancing effects of IL-4 in combination with TNF-alpha on RCC support the view that Th2 cytokines may be required for productive immune responses against RCC.

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Figures

Fig. 1

Fig. 1

Phenotype of the RCC cell line A-498. A-498 cells were subjected to flow cytometric analyses using antibodies against markers of RCC (G250) and epithelial cell-associated antigens. MHC class II, which is expressed exclusively on antigen-presenting cells, served as a negative control

Fig. 2

Fig. 2

Effects of IL-4 on the proliferation of A-498 cells: serum-dependence. A-498 cells were cultured in RPMI 1640 with or without serum supplemementation as indicated or in the chemically defined, serum-free culture medium AIM-V. Proliferation as assessed by [3H]-thymidine incorporation was determined in the presence or absence of IL-4 (1,000 U/ml). Results are mean values ± SEM of five independent experiments

Fig. 3

Fig. 3

Growth-modulatory effects of IL-4 on RCC in the context of pro-inflammatory cytokines. A-498 cells (a) or primary RCC cells (b) were cultured in the chemically defined, serum-free culture medium AIM-V. Proliferation as assessed by [3H]-thymidine incorporation was determined in the presence or absence of IL-4 (1,000 U/ml), TNF-α (1,000 U/ml), IL-1ß (10 ng/ml) or combinations. FACS histogram demonstrating G250 expression on primary RCC (b). Results are mean values of triplicate measurements ± SD. One out of three (a) and two experiments (b) with consistent results is shown

Fig. 4

Fig. 4

Induction of apoptosis in A-498 cells by IL-4 and TNF-α. A-498 cells were cultured in the chemically defined, serum-free culture medium AIM-V in the presence or absence of IL-4 (1,000 U/ml) or TNF-α (1,000 U/ml) or a combination of IL-4 and TNF-α. The percentage of apoptotic cells was determined by flow cytometry using the fluoresecent DNA intercalator 7-AAD. One out two experiments with nearly identical results is shown

Fig. 5

Fig. 5

Cytokine production by A-498 cells in response to IL-4 and TNF-α. A-498 cells in serum-free medium (AIM-V) were either left untreated (control) or treated with IL-4 (1,000 U/ml), TNF-α (1,000 U/ml) or a combination of IL-4 and TNF-α. Cytokine protein array (Raybio) was performed with the conditioned media. GRO (J1), IL-8 (J2), IL-6 (H2), MCP-1 (E3), M-CSF (H3), RANTES (B4), TNF-α (G4), Eotaxin (I5), FGF-4 (A6), GDNF (H6), HGF (I6), NAP-2 (J7), TIMP-1 (G8), TIMP-2 (H8)

Fig. 6

Fig. 6

Regulation of RANTES and MCP-1 production in A-498 cell by IL-4 and TNF-α. A-498 cells in serum-free medium (AIM-V) were either left untreated (control) or treated with IL-4 (1,000 U/ml), TNF-α (1,000 U/ml) or a combination of IL-4 and TNF-α. Cytokine multiplex analysis was performed with the conditioned media using the commercial Fluorokine MAP kits for RANTES and MCP-1 as well as the Luminex 100 analyzer

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References

    1. Motzer RJ, Bander NH, Nanus DM. Renal-cell carcinoma. N Engl J Med. 1996;335:865. doi: 10.1056/NEJM199609193351207. - DOI - PubMed
    1. Motzer RJ, Bacik J, Mazumdar M. Prognostic factors for survival of patients with stage IV renal cell carcinoma: Memorial Sloan-Kettering Cancer Center experience. Clin Cancer Res. 2004;10:6302S. doi: 10.1158/1078-0432.CCR-040031. - DOI - PubMed
    1. Thurnher M, Radmayr C, Hobisch A, Bock G, Romani N, Bartsch G, Klocker H. Tumor-infiltrating T lymphocytes from renal-cell carcinoma express B7–1 (CD80): T-cell expansion by T-T cell co-stimulation. Int J Cancer. 1995;62:559. doi: 10.1002/ijc.2910620512. - DOI - PubMed
    1. Ikemoto S, Yoshida N, Narita K, Wada S, Kishimoto T, Sugimura K, Nakatani T. Role of tumor-associated macrophages in renal cell carcinoma. Oncol Rep. 2003;10:1843. - PubMed
    1. Thurnher M, Radmayr C, Ramoner R, Ebner S, Bock G, Klocker H, Romani N, Bartsch G. Human renal-cell carcinoma tissue contains dendritic cells. Int J Cancer. 1996;68:1. doi: 10.1002/(SICI)1097-0215(19960927)68:1<1::AID-IJC1>3.0.CO;2-V. - DOI - PubMed

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