Experimental infection of dromedaries with Middle East respiratory syndrome-Coronavirus is accompanied by massive ciliary loss and depletion of the cell surface receptor dipeptidyl peptidase 4

Ann-Kathrin Haverkamp,Wolfgang Baumgärtner,Bart L Haagmans,Joaquim Segalés,Albert D M E Osterhaus,Judith M A Van Den Brand,Julia Vergara-Alert,Annika Lehmbecker,Ingo Spitzbarth,Albert Bensaid,Widagdo Widagdo

doi:10.1038/s41598-018-28109-2

Abstract

Middle East respiratory syndrome (MERS) represents an important respiratory disease accompanied by lethal outcome in one-third of human patients. Recent data indicate that dromedaries represent an important source of infection, although information regarding viral cell tropism and pathogenesis is sparse. In the current study, tissues of eight dromedaries receiving inoculation of MERS-Coronavirus (MERS-CoV) after recombinant Modified-Vaccinia-Virus-Ankara (MVA-S)-vaccination (n = 4), MVA-vaccination (mock vaccination, n = 2) and PBS application (mock vaccination, n = 2), respectively, were investigated. Tissues were analyzed by histology, immunohistochemistry, immunofluorescence, and scanning electron microscopy. MERS-CoV infection in mock-vaccinated dromedaries revealed high numbers of MERS-CoV-nucleocapsid positive cells, T cells, and macrophages within nasal turbinates and trachea at day four post infection. Double immunolabeling demonstrated cytokeratin (CK) 18 expressing epithelial cells to be the prevailing target cell of MERS-CoV, while CK5/6 and CK14 expressing cells did not co-localize with virus. In addition, virus was occasionally detected in macrophages. The acute disease was further accompanied by ciliary loss along with a lack of dipeptidyl peptidase 4 (DPP4), known to mediate virus entry. DPP4 was mainly expressed by human lymphocytes and dromedary monocytes, but overall the expression level was lower in dromedaries. The present study underlines significant species-specific manifestations of MERS and highlights ciliary loss as an important finding in dromedaries. The obtained results promote a better understanding of coronavirus infections, which pose major health challenges.

Highlights

In June 2012 a novel lineage C betacoronavirus (HCoV-EMC) was identified in a patient from the Kingdom of Saudi Arabia who suffered from acute pneumonia and renal failure[1]
Experimental Middle East respiratory syndrome (MERS)-CoV infection of dromedaries leads to high virus load in nasal turbinates and trachea accompanied by necro-suppurative inflammation at day 4 post infection
Experimentally infected dromedaries serve as an important animal model for investigations on certain aspects of MERS-CoV pathogenesis[21,22]

Summary

Introduction

In June 2012 a novel lineage C betacoronavirus (HCoV-EMC) was identified in a patient from the Kingdom of Saudi Arabia who suffered from acute pneumonia and renal failure[1]. In line with these observations, high amounts of MERS-CoV antigen were detected within the respiratory epithelium of the nasal turbinates of mock-vaccinated dromedaries at 4 dpi by immunohistochemistry in areas with most severe lesions (Fig. 2A).

Results

Conclusion