{
  "abstract": "Background Orosomucoid-like protein 3 (ORMDL3) has important role to regulate human rhino virus infection in airway epithelial cells as one of mechanisms of asthma exacerbations. In this report we created ORMDL3 knockout (KO) epithelial cell lines and tested the cells response to different stimulators and human rhinovirus 16 infections.Methods CRISPR/Cas9 techniques were applied to create ORMDL3 KOs in HEBC3-KT epithelial cells. Two targeted ORMDL3 KO clones were established. ORMDL3 KO cells were treated with different stimulators and human rhinovirus 16 (RV-A16). IL-6 and IL-8 levels, viral load, cytopathic effects were evaluated. ORMDL3 overexpression cells transfected by plasmids were also created to test with the stimulators.Results Two ORMDL3 KO clones with 1 base-pair deletion and 5 base-pair deletion in exon 2 were selected. Both deletions caused the frame shifts of ORMDL3, and the translations end prematurely. The knockout cells released more IL-6 and IL-8 after IL-1β, TNFα, Poly I:C and LPS/CD14 stimulations. With RV-A16 infection, ORMDL3 KO cells had less viral load, less cytopathic effects with higher level of IL-6 and IL-8 release. The effect on RV-16 infection could last 7 days in ORMDL3 KO cells after viral infection. Intercellular adhesion molecule 1 (ICAM-1) levels in KO cells were significantly increased with or without RV-16 infections.Conclusions We established ORMDL3 KO epithelial cells, which have shown to response to different stimuli. ORMDL3 KO cells significantly reduce RV-A16 infection. The ORMDL3 KO cells could provide a useful platform for investigating the mechanisms of human rhino virus infection in the mechanisms of asthma exacerbations.",
  "authors": [
    {
      "affiliations": [
        "National Heart and Lung Institute, Imperial College London, London, UK"
      ],
      "name": "Y Zhang"
    },
    {
      "affiliations": [
        "National Heart and Lung Institute, Imperial College London, London, UK"
      ],
      "name": "H Du"
    },
    {
      "affiliations": [
        "National Heart and Lung Institute, Imperial College London, London, UK"
      ],
      "name": "H Guo"
    },
    {
      "affiliations": [
        "National Heart and Lung Institute, Imperial College London, London, UK"
      ],
      "name": "L Daly"
    },
    {
      "affiliations": [
        "National Heart and Lung Institute, Imperial College London, London, UK"
      ],
      "name": "J Wong"
    },
    {
      "affiliations": [
        "National Heart and Lung Institute, Imperial College London, London, UK"
      ],
      "name": "M Wang"
    },
    {
      "affiliations": [
        "Department of Surgery and Cancer, Imperial College London, London, UK"
      ],
      "name": "E Chang"
    },
    {
      "affiliations": [
        "Department of Surgery and Cancer, Imperial College London, London, UK"
      ],
      "name": "D Ma"
    },
    {
      "affiliations": [
        "National Heart and Lung Institute, Imperial College London, London, UK"
      ],
      "name": "K Ito"
    },
    {
      "affiliations": [
        "National Heart and Lung Institute, Imperial College London, London, UK"
      ],
      "name": "WO Cookson"
    },
    {
      "affiliations": [
        "National Heart and Lung Institute, Imperial College London, London, UK"
      ],
      "name": "MF Moffatt"
    }
  ],
  "title": "P249 Knockout of ORMDL3 renders airway epithelial cell resistance to human rhinovirus infection",
  "uid": "96a77c48-c012-5de5-8887-1db28dd70dc4"
}