Adaptation of human enteric coronavirus to growth in cell lines

J. Ames P Luby, R. Ebecca Clinton, S. Tanley Kurtz

Research output: Contribution to journalArticle

10 Citations (Scopus)

Abstract

Background: The existence of human enteric coronavirus (HEC) has been debated since its first description in stool by electron microscopy (EM) in 1975. Needed to resolve the issue is its cultivation in readily available cell lines. Objectives: To grow HEC in cell lines. To describe its characteristics and to differentiate it from other human and animal coronaviruses. Study design: Originally grown in human fetal intestinal organ culture, HEC was passed in J774 cells (a mouse macrophage cell line) and C6/36 cells (a mosquito cell line). Its cytopathic effect (CPE) and pattern of immunofluorescence were described. Its appearance was ascertained by negative staining and transmission EM. Its structural proteins were delineated by polyacrylamide gel electrophoresis (PAGE) and Western blotting (WB). The antigenic character of the virus was determined by immunofluorescence and WB. Agglutination with mouse erythrocytes was performed. Results: In J774 cells, HEC induced the formation of giant cells and small syncytia. Immunofluorescence in both J774 and C6/36 cells was limited to the cytoplasm. Studies with transmission EM revealed the virus to have the typical appearance of other coronaviruses, to be 80-120 nm in diameter, and to bud into cysternae of the endoplasmic reticulum. By PAGE and WB, its major protein has an average molecular weight (MW) of 41 kilodaltons (kDa). Two other proteins had MWs of 190 and 24 kDa. By immunofluorescence and WB, HEC is antigenically distinct from human coronaviruses OC43 and 229E and mouse hepatitis virus (A59 strain). Preparations of HEC did not agglutinate mouse erythrocytes. Conclusion: We conclude that HEC is a human coronavirus that is antigenically unrelated to OC43 and 229E viruses. Growth of HEC in readily available cell lines should aid in elucidating its role as a pathogen in human diarrheal illnesses.

Original languageEnglish (US)
Pages (from-to)43-51
Number of pages9
JournalJournal of Clinical Virology
Volume12
Issue number1
DOIs
StatePublished - Jan 1999

Fingerprint

Coronavirus
Cell Line
Growth
Fluorescent Antibody Technique
Western Blotting
Giant Cells
Viruses
Transmission Electron Microscopy
Polyacrylamide Gel Electrophoresis
Human Coronavirus OC43
Human Coronavirus 229E
Erythrocytes
Murine hepatitis virus
Negative Staining
Proteins
Organ Culture Techniques
Agglutination
Culicidae
Endoplasmic Reticulum
Electron Microscopy

Keywords

  • Adaptation
  • C6/36 cells
  • Growth in cell lines
  • Human enteric coronavirus
  • J774 cells

ASJC Scopus subject areas

  • Applied Microbiology and Biotechnology
  • Virology
  • Immunology and Allergy
  • Infectious Diseases

Cite this

Adaptation of human enteric coronavirus to growth in cell lines. / Luby, J. Ames P; Clinton, R. Ebecca; Kurtz, S. Tanley.

In: Journal of Clinical Virology, Vol. 12, No. 1, 01.1999, p. 43-51.

Research output: Contribution to journalArticle

Luby, J. Ames P ; Clinton, R. Ebecca ; Kurtz, S. Tanley. / Adaptation of human enteric coronavirus to growth in cell lines. In: Journal of Clinical Virology. 1999 ; Vol. 12, No. 1. pp. 43-51.
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