Cytokine levels correlate with immune cell infiltration after anti-VEGF therapy in preclinical mouse models of breast cancer

• Published in: PloS one 2009-11, Vol.4 (11), p.e7669-e7669
• Main Authors: Roland, Christina L, Lynn, Kristi D, Toombs, Jason E, Dineen, Sean P, Udugamasooriya, D Gomika, Brekken, Rolf A
• Format: Article
• Language: English
• Subjects: Angiogenesis; Animal models; Animals; Binding sites; Biomarkers; Biomarkers, Tumor - metabolism; Breast cancer; Breast Neoplasms - blood; Breast Neoplasms - immunology; Cancer; Comparative analysis; Cytokines; Cytokines - metabolism; Dendritic cells; Disease Models, Animal; Electrochemiluminescence; Enzyme-linked immunosorbent assay; Female; Genetic engineering; Humans; Immune system; Immune System - immunology; Immunohistochemistry; Immunology/Immunomodulation; Immunology/Leukocyte Activation; Infiltration; Inflammation; Inhibition; Interleukin 6; Interleukin-1beta - blood; Interleukin-6 - blood; Kinases; Lymphatic system; Lymphocytes T; Macrophages; Medical prognosis; Metastases; Mice; Mice, Inbred BALB C; Mice, Inbred NOD; Mice, Transgenic; Neoplasm Transplantation; Neovascularization, Pathologic; Neutrophils; Oncology; Oncology/Breast Cancer; Solid tumors; Studies; Surgery; Therapy; Transgenic mice; Tumors; Vascular endothelial growth factor; Vascular Endothelial Growth Factor A - antagonists & inhibitors; Xenografts
• ISSN: 1932-6203; 1932-6203
• DOI: 10.1371/journal.pone.0007669

The effect of blocking VEGF activity in solid tumors extends beyond inhibition of angiogenesis. However, no studies have compared the effectiveness of mechanistically different anti-VEGF inhibitors with respect to changes in tumor growth and alterations in the tumor microenvironment. In this study we use three distinct breast cancer models, a MDA-MB-231 xenograft model, a 4T1 syngenic model, and a transgenic model using MMTV-PyMT mice, to explore the effects of various anti-VEGF therapies on tumor vasculature, immune cell infiltration, and cytokine levels. Tumor vasculature and immune cell infiltration were evaluated using immunohistochemistry. Cytokine levels were evaluated using ELISA and electrochemiluminescence. We found that blocking the activation of VEGF receptor resulted in changes in intra-tumoral cytokine levels, specifically IL-1beta, IL-6 and CXCL1. Modulation of the level these cytokines is important for controlling immune cell infiltration and ultimately tumor growth. Furthermore, we demonstrate that selective inhibition of VEGF binding to VEGFR2 with r84 is more effective at controlling tumor growth and inhibiting the infiltration of suppressive immune cells (MDSC, Treg, macrophages) while increasing the mature dendritic cell fraction than other anti-VEGF strategies. In addition, we found that changes in serum IL-1beta and IL-6 levels correlated with response to therapy, identifying two possible biomarkers for assessing the effectiveness of anti-VEGF therapy in breast cancer patients.