Cellular entry of hantaviruses which cause hemorrhagic fever with renal syndrome is mediated by β3 integrins

Hantaviruses replicate primarily in the vascular endothelium and cause two human diseases, hemorrhagic fever with renal syndrome (HFRS) and hantavirus pulmonary syndrome (HPS). In this report, we demonstrate that the cellular entry of HFRS-associated hantaviruses is facilitated by specific integrins expressed on platelets, endothelial cells, and macrophages. Infection of human umbilical vein endothelial cells and Vero E6 cells by the HFRS-causing hantaviruses Hantaan (HTN), Seoul (SEO), and Puumala (PUU) is inhibited by antibodies to alpha(v)beta(3) integrins and by the integrin ligand vitronectin. The cellular entry of HTN, SEO, and PUU viruses, but not the nonpathogenic Prospect Hill (PH) hantavirus (i.e., a virus with no associated human disease), was also mediated by introducing recombinant alpha(IIb)beta(3) or alpha(v)beta(3) integrins into beta(3)-integrin-deficient CHO cells. In addition, PH infectivity was not inhibited by alpha(v)beta(3)-specific sera or vitronectin but was blocked by alpha(5)beta(1)-specific sera and the integrin ligand fibronectin. RGD tripeptides, which are required for many integrin-ligand interactions, are absent from all hantavirus G1 and G2 surface glycoproteins, and GRGDSP peptides did not inhibit hantavirus infectivity. Further, a mouse-human hybrid beta(3) integrin-specific Fab fragment, c7E3 (ReoPro), also inhibited the infectivity of HTN, SEO, and PUU as well as HPS-associated hantaviruses, Sin Nombre (SN) and New York-1 (NY-1). These findings indicate that pathogenic HPS- and HFRS-causing hantaviruses enter cells via beta(3) integrins, which are present on the surfaces of platelets, endothelial cells, and macrophages. Since beta(3) integrins regulate vascular permeability and platelet function, these findings also correlate beta(3) integrin usage with common elements of hantavirus pathogenesis.