Supplementary Figures 1-4 with figure legends from Differential Expression of Homing Receptor Ligands on Tumor-Associated Vasculature that Control CD8 Effector T-cell Entry

Woods, Amber N.; Wilson, Ashley L.; Srivinisan, Nithya; Zeng, Jianhao; Dutta, Arun B.; Peske, J. David; Tewalt, Eric F.; Gregg, Randal K.; Ferguson, Andrew R.; Engelhard, Victor H.

doi:10.1158/2326-6066.22538255.v1

Supplementary Figures 1-4 with figure legends from Differential Expression of Homing Receptor Ligands on Tumor-Associated Vasculature that Control CD8 Effector T-cell Entry

https://doi.org/10.1158/2326-6066.22538255

journal contribution

posted on 2023-04-03, 23:08 authored by Amber N. Woods, Ashley L. Wilson, Nithya Srivinisan, Jianhao Zeng, Arun B. Dutta, J. David Peske, Eric F. Tewalt, Randal K. Gregg, Andrew R. Ferguson, Victor H. Engelhard

<p>S1: LLC-OVA or MC-38-OVA tumors were harvested on day 14 (SC) or day 11 (IP). LLC-OVA tumors were flash frozen and stained for MadCAM-1 expression. Representative and summary data interrogating 8-10 random fields from single sections of 3 tumors each for the percent of CD31+ pixels that were MadCAM-1+. Percentages were calculated using ImageJ to define CD31+ pixels and quantitating those positive for MadCAM-1. MC-38-OVA tumors were digested and stained for flow cytometry. Summary data (n=2 tumors per group) for the percent of CD31+CD45+ cells that expressed E-Selectin. Statistical analysis performed using unpaired student's T-test. S2: B16-OVA tumors were harvested on day 21 (Lung) or day 13 (Brain) and dissociated and stained for fow cytometric analysis. Representative and summary data for expression of VCAM-1, E-selectin, and MadCAM-1 endothelial cells isolated from lung and brain tumors (n=4). Plots are gated on CD31+CD45- cells. S3: B16-OVA and B16-F1 tumors were harvested on day 14 (SC) or day 11 (IP) and flash frozen for immunofluorescence staining with anti-CD8. Summary data interrogating 5 random 200X fields from single sections of 2 tumors to assess the number of CD8 T-cells. Statistical analysis performed using unpaired student's T-test. 4: CD8 effector T-cells were generated using SIINFEKL pulsed BMDCs administered IV, IP, or SC as described in the legends to Figures 4 and 5. Freshly isolated ex vivo effectors were stained with the indicated antibodies and analyzed by flow cytometry. Plots are gated on CD45+CD8+Thy1.1+ cells.</p>

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American Cancer Society

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DOI - Is supplement to Differential Expression of Homing Receptor Ligands on Tumor-Associated Vasculature that Control CD8 Effector T-cell Entry

ARTICLE ABSTRACT

Although CD8+ T cells are critical for controlling tumors, how they are recruited and home to primary and metastatic lesions is incompletely understood. We characterized the homing receptor (HR) ligands on tumor vasculature to determine what drives their expression and their role in T-cell entry. The anatomic location of B16-OVA tumors affected the expression of E-selectin, MadCAM-1, and VCAM-1, whereas the HR ligands CXCL9 and ICAM-1 were expressed on the vasculature regardless of location. VCAM-1 and CXCL9 expression was induced by IFNγ-secreting adaptive immune cells. VCAM-1 and CXCL9/10 enabled CD8+ T-cell effectors expressing α4β1 integrin and CXCR3 to enter both subcutaneous and peritoneal tumors, whereas E-selectin enabled E-selectin ligand+ effectors to enter subcutaneous tumors. However, MadCAM-1 did not mediate α4β7+ effector entry into peritoneal tumors due to an unexpected lack of luminal expression. These data establish the relative importance of certain HRs expressed on activated effectors and certain HR ligands expressed on tumor vasculature in the effective immune control of tumors. Cancer Immunol Res; 5(12); 1062–73. ©2017 AACR.