IFN Effects on Lymphatic Endothelial Cells

Primary mLECs (Cell Biologics, C57–6092) and hLECs (Cell Biologics, H-6092) were cultured in in endothelial cell media (Cell biologics, M1168 and H1168). Flasks and plates were coated with matrigel diluted 1:1000 in media(31 (link)). mLECS were treated with rIFNα (a kind gift from Ross Kedl(32 (link))) and rIFNγ (Biolegend, 714006) at 250 U/mL(33 (link)) for 24 hours (h) before cells were harvested. Cells were then stained with αPDPN and αPD-L1, and acquired on a Cyan ADP flow cytometer (Dako) and analyzed using flowjo software (Treestar). hLECs were treated with human IFNα2 (PBL Assay Science-11100–1) at 500 U/mL(34 (link)) for 24 h and then harvested as above.

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Lucas E.D., Finlon J.M., Burchill M.A., McCarthy M.K., Morrison T.E., Colpitts T.M, & Jirón Tamburini B.A. (2018). Type 1 interferon and PD-L1 coordinate lymphatic endothelial cell expansion and contraction during an inflammatory immune response. Journal of immunology (Baltimore, Md. : 1950), 201(6), 1735-1747.

Publication 2018

C57–6092 M1168 H1168 rIFNγ Cyan ADP flow cytometer human IFNα2

Assay Biologics Cells Endothelial cell Human Matrigel

Corresponding Organization : University of Colorado Anschutz Medical Campus

Other organizations : Boston University

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Variable analysis

independent variables

Treatment of mLECs with rIFNα and rIFNγ at 250 U/mL for 24 hours
Treatment of hLECs with human IFNα2 at 500 U/mL for 24 hours

dependent variables

Expression of αPDPN and αPD-L1 in mLECs and hLECs, measured by flow cytometry

control variables

Cell culture conditions (endothelial cell media, matrigel-coated flasks and plates)
Culture duration (24 hours)

controls

Positive control: None specified
Negative control: None specified

Annotations

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