A CCR4 antagonist reverses the tumor-promoting microenvironment of renal cancer
Chiara Berlato, … , Sergio A. Quezada, Frances R. Balkwill
Chiara Berlato, … , Sergio A. Quezada, Frances R. Balkwill
Published January 30, 2017
Citation Information: J Clin Invest. 2017;127(3):801-813. https://doi.org/10.1172/JCI82976.
View: Text | PDF
Research Article Immunology Oncology

A CCR4 antagonist reverses the tumor-promoting microenvironment of renal cancer

Abstract

Elevated expression of the chemokine receptor CCR4 in tumors is associated with poor prognosis in several cancers. Here, we have determined that CCR4 was highly expressed in human renal cell carcinoma (RCC) biopsies and observed abnormal levels of CCR4 ligands in RCC patient plasma. An antagonistic anti-CCR4 antibody had antitumor activity in the RENCA mouse model of RCC. CCR4 inhibition did not reduce the proportion of infiltrating leukocytes in the tumor microenvironment but altered the phenotype of myeloid cells, increased NK cell and Th1 cytokine levels, and reduced immature myeloid cell infiltrate and blood chemokine levels. In spite of prominent changes in the myeloid compartment, the anti-CCR4 antibody did not affect RENCA tumors in T cell–deficient mice, and treatment with an anti–class II MHC antibody abrogated its antitumor activity. We concluded that the effects of the anti-CCR4 antibody required the adaptive immune system and CD4+ T cells. Moreover, CCL17-induced IFN-γ production was reduced when Th1-polarized normal CD4+ T cells were exposed to the CCR4 ligand, evidencing the involvement of CCR4 in Th1/Th2 regulation. The anti-CCR4 antibody, alone or in combination with other immune modulators, is a potential treatment approach to human solid cancers with high levels of CCR4-expressing tumor-infiltrating leukocytes and abnormal plasma CCR4 ligand levels.

Authors

Chiara Berlato, Moddasar N. Khan, Tiziana Schioppa, Richard Thompson, Eleni Maniati, Anne Montfort, Maryam Jangani, Monica Canosa, Hagen Kulbe, Urs B. Hagemann, Alexander R. Duncan, Laura Fletcher, Robert W. Wilkinson, Thomas Powles, Sergio A. Quezada, Frances R. Balkwill

×

Figure 3

Effects of anti-CCR4 on the RENCA TAMs.

Options: View larger image (or click on image) Download as PowerPoint
Effects of anti-CCR4 on the RENCA TAMs. BALB/c mice were injected with R...
BALB/c mice were injected with RENCA-luc cells and treated with Affi-5 (T) or isotype control (C). Mice were sacrificed 17 days after surgery, and tumors were dissociated and characterized by flow cytometry. (A) Tumor-infiltrating macrophages (gated as CD45+CD11b+F4/80+) per milligram of tumor for 5 independent experiments are shown. There was no significant difference between Affi-5–treated (T) and isotype-treated (C) tumors (n = 15 and n = 14, respectively). (B and C) Geometric mean of fluorescence intensity (MFI) for MHCII and MR staining on macrophages for 5 independent experiments, and staining for isotype-treated and Affi-5–treated dissociated tumors for 1 representative experiment. Two-tailed Student’s t test, ***P = 0.0008 and **P = 0.0085. n = 19 for C, n = 17 for T. (D) RNA was extracted from macrophages (CD45+CD11b+F4/80+) sorted by flow cytometry from dissociated tumors. The ratio between arginase and Nos2 expression was determined by real-time PCR in 2 independent experiments pooled together (Mann-Whitney U test, *P = 0.035), with n = 6 for C, n = 7 for T. (E) Cells were dissociated at end point from dissected tumors from BALB/c mice treated with Affi-5 (T) or isotype control (C) and plated overnight in the presence of brefeldin A. The fold change in the number of macrophages (CD45+CD11b+F4/80+) positive for intracellular TNF-α is shown from 2 pooled experiments (Mann-Whitney U test, **P = 0.002, n = 11 for C and n = 15 for T).