Journal: Oncoimmunology

Article Title: TIE-2-expressing monocytes are lymphangiogenic and associate specifically with lymphatics of human breast cancer

doi: 10.1080/2162402X.2015.1073882

Figure Lengend Snippet: In human breast tumors, TEM can be identified solely based on CD14 immunostaining. (A) Immunofluorescence staining and confocal analysis of human BC tumor sections showing CD14, TIE-2 and VEGFR-1 triple positive cells. Representative image from 16 patients; (B) CD14 and TIE-2 expression correlation analysis. The dot plot (left panel) represents all tiles for one representative image; the box plot (right panel) shows the Spearman’s rank correlation coefficient (ρ) for CD14 and TIE-2 expression in 11 patients. In the dot plot, 54% of CD14 + tiles and 70% of TIE-2 + tiles were above the threshold. 100% of CD14 + tiles were TIE-2 + which means that the high correlation (ρ = 0.844) applied to all CD14 + tiles; (C) CD14 and VEGFR-1 expression correlation analysis for 11 patients. In the dots plot, 55% of CD14 + tiles and 75% of VEGFR-1 + tiles were above the threshold. 100% of CD14 + tiles are VEGFR-1 + which means that the high correlation (ρ = 0.852) applied to all CD14 + tiles; (D) Example of TIE-2 expression in TEM, clusters of BEC (arrow heads) and blood vessels (right panel) shown in confocal microscopy images of BC sections. TEM and blood endothelial structures were stained with CD14 and CD31, respectively; (E) Example of immunofluorescence labeling of CD14, VEGFR-1 and TIE-2 in sections containing non-neoplastic breast tissue adjacent to tumor tissues. CD14 + cells (arrows) are VEGFR - and TIE-2 - . Non-neoplastic (panel E) and tumor tissue (panel A) were stained and imaged simultaneously under the same conditions and thus intensities of the expression of TIE-2 and VEGFR-1 signals can be compared. Representative images from four patients. Scale bars: 25 μm.

Article Snippet: TIE-2 inhibitor compound 7 was from Alexis Biochemicals (San Diego, CA) and VEGFR inhibitor PTK787 from Novartis Institutes for BioMedical Research (Basel, Switzerland).

Techniques: Immunostaining, Immunofluorescence, Staining, Expressing, Confocal Microscopy, Labeling

Journal: Oncoimmunology

Article Title: TIE-2-expressing monocytes are lymphangiogenic and associate specifically with lymphatics of human breast cancer

doi: 10.1080/2162402X.2015.1073882

Figure Lengend Snippet: TEM lymphangiogenic activity is supported by cross-talk between VEGFR and TIE-2 pathways. (A) In vivo corneal vascularization assay was used to assess the lymphangiogenic activity of TEM isolated from human breast tumors. Lymphatic vessels emerging from the peripheral limbal vasculature were labeled with LYVE-1-specific antibodies in sagittal sections of mouse eyes. Double-headed and single arrows depict cornea and iris respectively. Scale bar: 100 μm; (B) Confocal microscopy images of TEM sorted from dissociated breast tumor showing expression of VEGF-A, -C and -D. Scale bar: 5μm; (C) TEM isolated from dissociated breast tumors were exposed to TIE-2- and VEGFR-specific kinase inhibitors and their hem- and lymphangiogenic activities were measured in vitro using a sprouting assay of HUVEC and LEC respectively ( t test, *** p <0.001, **** p < 0.0001).

Article Snippet: TIE-2 inhibitor compound 7 was from Alexis Biochemicals (San Diego, CA) and VEGFR inhibitor PTK787 from Novartis Institutes for BioMedical Research (Basel, Switzerland).

Techniques: Activity Assay, In Vivo, Vascularization Assay, Isolation, Labeling, Confocal Microscopy, Expressing, In Vitro