Endothelial STING controls T cell transmigration in an IFNI-dependent manner
Marina Anastasiou, Gail A. Newton, Kuljeet Kaur, Francisco J. Carrillo-Salinas, Sasha A. Smolgovsky, Abraham L. Bayer, Vladimir Ilyukha, Shruti Sharma, Alexander Poltorak, Francis W. Luscinskas, Pilar Alcaide
Marina Anastasiou, Gail A. Newton, Kuljeet Kaur, Francisco J. Carrillo-Salinas, Sasha A. Smolgovsky, Abraham L. Bayer, Vladimir Ilyukha, Shruti Sharma, Alexander Poltorak, Francis W. Luscinskas, Pilar Alcaide
View: Text | PDF
Research Article Inflammation Vascular biology

Endothelial STING controls T cell transmigration in an IFNI-dependent manner

Abstract

The stimulator of IFN genes (STING) protein senses cyclic dinucleotides released in response to double-stranded DNA and functions as an adaptor molecule for type I IFN (IFNI) signaling by activating IFNI-stimulated genes (ISG). We found impaired T cell infiltration into the peritoneum in response to TNF-α in global and EC-specific STING–/– mice and discovered that T cell transendothelial migration (TEM) across mouse and human endothelial cells (EC) deficient in STING was strikingly reduced compared with control EC, whereas T cell adhesion was not impaired. STING–/– T cells showed no defect in TEM or adhesion to EC, or immobilized endothelial cell–expressed molecules ICAM1 and VCAM1, compared with WT T cells. Mechanistically, CXCL10, an ISG and a chemoattractant for T cells, was dramatically reduced in TNF-α–stimulated STING–/– EC, and genetic loss or pharmacologic antagonisms of IFNI receptor (IFNAR) pathway reduced T cell TEM. Our data demonstrate a central role for EC-STING during T cell TEM that is dependent on the ISG CXCL10 and on IFNI/IFNAR signaling.

Authors

Marina Anastasiou, Gail A. Newton, Kuljeet Kaur, Francisco J. Carrillo-Salinas, Sasha A. Smolgovsky, Abraham L. Bayer, Vladimir Ilyukha, Shruti Sharma, Alexander Poltorak, Francis W. Luscinskas, Pilar Alcaide

×

Figure 3

Decreased T cell recruitment into the peritoneal cavity of EC-STING–/– mice in response to TNF-α.

Options: View larger image (or click on image) Download as PowerPoint
Decreased T cell recruitment into the peritoneal cavity of EC-STING–/– m...
(A) Schematic gene-targeting map of STING gene showing STING floxed and STING conditional alleles before and after tamoxifen (TMX) treatment (75 mg/kg body weight) and primer (P1 and P2) binding sites, with orange arrows pointing at the LoxP sites. (B) Primer pair P1 and P2 were used to detect unexcised and excised STING alleles in the heart (H), Splenocytes (Sp), MHEC (EC), and T cells (T) purified from Cad5ERTCre2+/– STINGfl/fl treated with vehicle or TMX. (C) Quantification of STING protein expression in cultured MHEC from Cad5ERTCre2+/– STINGfl/fl and WT Th1 cells treated with vehicle or 4OH-TMX for 24 h. (D) Western blotting images of 4OH-TMX–treated cell lysate. Each line is an independent cell preparation (n = 3). (E) Representative flow cytometric panels of CD45+CD4+ cells recruited to the peritoneal cavity 24 h after TNF-α. (F and G)Quantification of total CD45+ (F) and CD4+ (G) cells recruited cells to the peritoneal cavity 24 h after TNF-α. (H) Representative flow cytometric panels of CD45+Gr1+ cells recruited to the peritoneal cavity 4 h after TNF-α. (I and J) Quantification of total CD45+ (I) and Gr1+ (J) cells recruited to the peritoneal cavity 4 h after TNF-α. Data represent n = 2 independent experiments; n = 3 control and n = 4 TMX-treated mice per experiment (24 h TNF-α.); and n = 6 animals per group (4 h TNF-α). *P < 0.05, **P < 0.01 and ***P < 0.001; t test.