The HeLa cell receptor for enterovirus 70 is decay-accelerating factor (CD55)

Abstract

Enterovirus 70 (EV70) is a recently emerged human pathogen belonging to the family Picornaviridae. The ability of EV70 to infect a wide variety of nonprimate cell lines in vitro is unique among human enteroviruses. The importance of virus receptors as determinants of viral host range and tropism led us to study the host cell receptor for this unusual picornavirus. We produced a monoclonal antibody (MAb), EVR1, which bound to the surface of HeLa cells and protected them against infection by EV70 but not by poliovirus or by coxsackievirus B3. This antibody also inhibited the binding of [ 35 S]EV70 to HeLa cells. MAb EVR1 did not bind to monkey kidney (LLC-MK 2) cells, nor did it protect these cells against virus infection. In Western immunoassays and in immunoprecipitations, MAb EVR1 identified a HeLa cell glycoprotein of approximately 75 kDa that is attached to the cell membrane by a glycosyl-phosphatidylinositol (GPI) anchor. Decay-accelerating factor (DAF, CD55) is a 70-to 75-kDa GPI-anchored membrane protein that is involved in the regulation of complement and has also been shown to function as a receptor for several enteroviruses. MAb EVR1 bound to Chinese hamster ovary (CHO) cells constitutively expressing human DAF. Anti-DAF MAbs inhibited EV70 binding to HeLa cells and protected them against EV70 infection. Transient expression of human DAF in murine NIH 3T3 cells resulted in binding of labelled EV70, and stably transformed NIH 3T3 cells expressing DAF were able to support virus replication. These data indicate that the HeLa cell receptor for EV70 is DAF.

Blood Groups in Infection and Host Susceptibility

“…ENV70 was shown to bind HeLa cells via CD55. Epitope mapping with anti-CD55 MAbs suggested binding to CCP1, -2, and -3 (653,654). Experiments with DAF deletion constructs localized binding primarily to CCP1.…”

Glycosyl-phosphatidylinositol (GPI)-anchored membrane association of the porcine reproductive and respiratory syndrome virus GP4 glycoprotein and its co-localization with CD163 in lipid rafts

“…A GPI-anchored protein can be distinguished from its un-modified form by its mobility (Ikezawa, 2002), and thus the migration pattern of GP4 was examined by SDS-PAGE and western blot. HeLa cells naturally express human DAF as a 41 kDa protein before GPImodification (Karnauchow et al, 1996) and after glycosylation and GPI-modification, it becomes approximately 82-88 kDa (Nickells et al, 1994), which can readily be detectable by DAF MAb from cell lysates (Fig. 2B, lane 4).…”

“…Monoclonal antibody (MAb) against human DAF ), anti-FLAG MAb (clone M2), and anti-FLAG rabbit antibody were from Sigma-Aldrich (St. Louis, MO); anti-GFP rabbit antibody from Cell Signaling Technology (Danvers, MA); β-actin MAb (AC-15) from Santa Cruz Biotechnology (Santa Cruz, CA); and anti-porcine CD163 MAb (clone 2A100/11) from AbD Serotec (Raleigh, NC). Tissue culture supernatant containing MAb EVR1 raised against human DAF was kindly provided by K. Dimmock (University of Ottawa, Ottawa, Canada) and described elsewhere (Karnauchow et al, 1996). Alexa Fluor 488®-conjugated goat anti-mouse IgG (H + L), Alexa Fluor 594®-conjugated goat anti-rabbit IgG (H+ L), and G418 (neomycin sulfate analog) were purchased from Invitrogen (Carlsbad, CA).…”

Evolution of Cell Recognition by Viruses: A Source of Biological Novelty with Medical Implications

“…Integrin v 3 (Nelsen-Salz et al, 1999) Echovirus 3, 6, 7, 11-13, 20, 21, 24, 29, 33 DAF (Bergelson et al, 1994;Ward et al, 1994;Powell et al, 1997); MHC class I via 2 -microglobulin (Ward et al, 1998); HS-GAG (Goodfellow et al, 2001); CD59 (Goodfellow et al, 2000) Enterovirus 70 DAF (Karnauchow et al, 1996) TMEV strains BeAn, DA Sialic acid (Zhou et al, , 2000 TMEV strain GDVII UDP-galactose transporter (or galactose containing glycoprotein) (Hertzler et al, 2001); HS-GAG (Reddi and Lipton, 2002) Poliovirus 1-3 PVR (Mendelsohn et al, 1989;Racaniello, 1996;Belnap et al, 2000;He et al, 2000) Swine vesicular disease virus CAR (Martino et al, 2000)…”