Cell cycle–dependent expression of CXC chemokine receptor 3 by endothelial cells mediates angiostatic activity
Paola Romagnani, … , Sergio Romagnani, Mario Serio
Paola Romagnani, … , Sergio Romagnani, Mario Serio
Published January 1, 2001
Citation Information: J Clin Invest. 2001;107(1):53-63. https://doi.org/10.1172/JCI9775.
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Article

Cell cycle–dependent expression of CXC chemokine receptor 3 by endothelial cells mediates angiostatic activity

Abstract

Endothelial cell receptors for the angiostatic chemokines IFN-γ–inducible protein of 10 kDa (IP-10) and monokine induced by IFN-γ (Mig) have not yet been identified, and the mechanisms responsible for the effects of these chemokines on angiogenesis are still unclear. IP-10 and Mig share a common functional receptor on activated T lymphocytes, named CXC chemokine receptor 3 (CXCR3). Using in situ hybridization and immunohistochemistry, we show that CXCR3 is expressed by a small percentage of microvascular endothelial cells in several human normal and pathological tissues. Primary cultures of human microvascular endothelial cells (HMVECs) likewise express CXCR3, although this expression is limited to the S/G2-M phase of their cell cycle. Both IP-10 and Mig, as well as the IFN-γ–inducible T-cell α chemoattractant (I-TAC), which all share high-affinity binding for CXCR3, block HMVEC proliferation in vitro, an effect that can be inhibited by an anti-CXCR3 antibody. These data provide definitive evidence of CXCR3 expression by HMVEC and open new avenues for therapeutic interventions in all conditions in which an angiostatic effect may be beneficial.

Authors

Paola Romagnani, Francesco Annunziato, Laura Lasagni, Elena Lazzeri, Chiara Beltrame, Michela Francalanci, Mariagrazia Uguccioni, Grazia Galli, Lorenzo Cosmi, Lucia Maurenzig, Marco Baggiolini, Enrico Maggi, Sergio Romagnani, Mario Serio

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Figure 3

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CXCR3 mRNA and protein expression by primary cultures of HMVEC. (a) Tota...
CXCR3 mRNA and protein expression by primary cultures of HMVEC. (a) Total RNA from a Th1 clone, HMCs, and HMVECs was reverse transcribed and PCR amplified using CXCR3 (top) and β-actin (bottom) primers, as described in Methods. An aliquot of each PCR reaction was electrophoresed on 1.5% agarose gel and visualized with ethidium bromide. An incubation performed in the absence of RT is also shown (no RT). (b) Western blot analysis of extracts from activated Th1 cells, HMCs, and HMVECs. Cell lysates were separated on SDS-PAGE under nonreducing conditions, transferred to nitrocellulose membranes, and assessed with the 49801.111 anti-CXCR3 mAb. (c) Flow cytometry of CXCR3 expression, evaluated with two different anti-CXCR3 mAb’s (1C6, dashed line; 498011.111, solid line), on Th1 cells, HMCs, and HMVECs. The black peak represents staining of the same cells with the isotype-matched control antibody. One of eight separate experiments for each technique is shown.