Comparison of the Replication of Distinct Strains of Human Coronavirus OC43 in Organotypic Human Colon Cells (Caco-2) and Mouse Intestine
Three strains of human Coronavirus (HCV) OC43 were compared for their ability to cause enteric infections and to induce interferon alpha (IFNα) using the Caco-2 human colon carcinoma cell line which exhibits spontaneous epithelial differentiation in vitro. MRC-5 cell culture grown stocks were prepared from: 1. CV Paris, a strain of 0C43 recovered from an outbreak of necrotizing enterocolitis in newborns. 2. CV Mb, a neurotropic strain of OC43 which exhibits strict neuronal specificity in murine neuronal cell cultures. 3. CV Rd, a strain of OC43 which grows to a high titer in human rhabdomyosarcoma (RD) cells. Immunofluorescent staining for nucleocapsid antigen and plaque assay in MRC-5 cells was used to detect viral replication. BG-9 (human foreskin) cells challenged with vesicular stomatitis virus were used to detect IFNα production by human peripheral blood monocytes (PBMC) stimulated bv virus infected Caco-2 cells.
Caco-2 cells infected with virus at a multiplicity of infection of 0.5 yielded 104.6 and 104.4 plaque forming units/ml (pfu/ml) with CV Rd and CV Paris respectively, while CV Mb yielded only 103 pfu/ml. Caco-2 cells infected with CV Rd induced 64 IU/ml of IFNα in PBMC while these cells infected with CV Paris induced <2 IU/ml IFNα. In cells infected with CV Mb 4 IU/ml IFNα was detected. The results suggest that a lack of IFNα induction by CV Paris may be an indicator of its enteropathogenic potential.
When newborn Balb/c mice were given 10 μ1 of CV Paris orally, viral antigens could be demonstrated in intestinal homogenates inoculated onto RC cells and an antibody titer of 40 at three weeks post infection was shown by immunodot blot. Some virus was recovered from mice given mouse neurotropic CV Mb strain orally but the serum antibody titer was <20. Therefore CV Paris but not CV Mb is enterotropic and induces immunity in mice given the virus orally.
KeywordsPeripheral Blood Mononuclear Cell Vesicular Stomatitis Virus Necrotizing Enterocolitis Human Colon Carcinoma Cell Line Plaque Reduction Assay
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