Use of SLAM and PVRL4 and identification of pro-HB-EGF as cell entry receptors for wild type phocine distemper virus

PLoS One. 2014 Aug 29;9(8):e106281. doi: 10.1371/journal.pone.0106281. eCollection 2014.

Abstract

Signalling lymphocyte activation molecule (SLAM) has been identified as an immune cell receptor for the morbilliviruses, measles (MV), canine distemper (CDV), rinderpest and peste des petits ruminants (PPRV) viruses, while CD46 is a receptor for vaccine strains of MV. More recently poliovirus like receptor 4 (PVRL4), also known as nectin 4, has been identified as a receptor for MV, CDV and PPRV on the basolateral surface of polarised epithelial cells. PVRL4 is also up-regulated by MV in human brain endothelial cells. Utilisation of PVRL4 as a receptor by phocine distemper virus (PDV) remains to be demonstrated as well as confirmation of use of SLAM. We have observed that unlike wild type (wt) MV or wtCDV, wtPDV strains replicate in African green monkey kidney Vero cells without prior adaptation, suggesting the use of a further receptor. We therefore examined candidate molecules, glycosaminoglycans (GAG) and the tetraspan proteins, integrin β and the membrane bound form of heparin binding epithelial growth factor (proHB-EGF),for receptor usage by wtPDV in Vero cells. We show that wtPDV replicates in Chinese hamster ovary (CHO) cells expressing SLAM and PVRL4. Similar wtPDV titres are produced in Vero and VeroSLAM cells but more limited fusion occurs in the latter. Infection of Vero cells was not inhibited by anti-CD46 antibody. Removal/disruption of GAG decreased fusion but not the titre of virus. Treatment with anti-integrin β antibody increased rather than decreased infection of Vero cells by wtPDV. However, infection was inhibited by antibody to HB-EGF and the virus replicated in CHO-proHB-EGF cells, indicating use of this molecule as a receptor. Common use of SLAM and PVRL4 by morbilliviruses increases the possibility of cross-species infection. Lack of a requirement for wtPDV adaptation to Vero cells raises the possibility of usage of proHB-EGF as a receptor in vivo but requires further investigation.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Animals
  • Antigens, CD / genetics
  • Antigens, CD / metabolism*
  • CHO Cells
  • Cell Adhesion Molecules / genetics
  • Cell Adhesion Molecules / metabolism*
  • Chlorocebus aethiops
  • Cricetinae
  • Cricetulus
  • Distemper / genetics
  • Distemper / metabolism
  • Distemper Virus, Phocine / physiology*
  • Dogs
  • Heparin-binding EGF-like Growth Factor / genetics
  • Heparin-binding EGF-like Growth Factor / metabolism*
  • Humans
  • Receptors, Cell Surface / genetics
  • Receptors, Cell Surface / metabolism*
  • Receptors, Virus / genetics
  • Receptors, Virus / metabolism*
  • Signaling Lymphocytic Activation Molecule Family Member 1
  • Vero Cells
  • Virus Internalization*

Substances

  • Antigens, CD
  • Cell Adhesion Molecules
  • Heparin-binding EGF-like Growth Factor
  • Receptors, Cell Surface
  • Receptors, Virus
  • Signaling Lymphocytic Activation Molecule Family Member 1

Grants and funding

Mary Melia was supported by a studentship from the Department of Education and Learning (DEL), Northern Ireland. This work was also funded by grant 9.1 from the Health and Social Care Research and Development Office Northern Ireland. The funders had no role in study design, data collection and analysis, decision to publish, or preparation of the manuscript.