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Epithelial Cell Extrusion Zones Observed on Confocal Laser Endomicroscopy Correlates with Immunohistochemical Staining of Mucosal Biopsy Samples

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Abstract

Background and Aims

The density of epithelial cell extrusion zones in the intestinal lining, also known as gap density (number of gaps/1000 epithelial cells counted), can be quantitated using probe-based confocal laser endomicroscopy (pCLE). Gap density has been reported to be higher than normal in both inflammatory bowel disease (IBD) and irritable bowel syndrome (IBS) patients. Epithelial cells destined for extrusion from the intestinal surface would stain positive for either activated caspase-1 or caspase-3 on mucosal biopsy samples. The aim of this study was to determine whether epithelial gap density on pCLE correlates with quantitative analysis of activated caspase staining of mucosal biopsy samples from patients.

Methods

We obtained pCLE images and biopsy samples of the terminal ileum during colonoscopies of healthy controls and patients with either IBD or IBS. The pCLE images and biopsy samples were blindly analyzed for gap density and for cells staining positive for activated caspases, respectively. The degree of correlation was determined using nonparametric statistical tests.

Results

The median results were 10 gaps/1000 cells counted for controls versus 33 gaps/1000 cells counted for chronic intestinal disorder patients (p = 0.02). Activated caspase staining showed 13 positive cells/1000 epithelial cells counted versus 26 positive cells/1000 epithelial cells counted, respectively (p = 0.02), thus showing a strong correlation with a Spearman’s coefficient ρ of 0.61 (strong correlation for ρ = 0.4–0.75, p = 0.01).

Conclusions

Intestinal epithelial gap density via pCLE correlated strongly with quantitative analysis of immunohistochemical staining of mucosal biopsy samples.

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Acknowledgments

We would like to thank Ms. Stephanie Mah and Mr. Harsh Thaker for their assistance with the experiments. Dr. Julia Liu is a recipient of the Canadian Institutes of Health Research New Investigator Salary award. Funding support for the study was provided by the Canadian Institutes of Health Research/Canadian Association of Gastroenterology (CIHR/CAG GNO 96692), the University of Alberta Hospital Foundation (UHF 2011), and NIH P20 GM103625 (Center for Microbial Pathogenesis and Host Inflammatory Responses at UAMS). Ms. Theresa Kay was a recipient of a University of Alberta Faculty of Medicine and Dentistry summer studentship.

Grant support

Grant support was provided by the Canadian Institute of Health Research New Investigator Salary Award (JJL) Canadian Association of Gastroenterology (CIHR/CAG GNO 96692 JJL), Crohn’s and Colitis Foundation of Canada (JJL), the University of Alberta Hospital Foundation (UHF 2011, JJL), National Institute of Health P20 (GM103625, JJL), Faculty of Medicine and Dentistry summer studentship (TMK), the Canadian Foundation for Innovations (RNF), and Natural Sciences and Engineering Research Council of Canada (RTI).

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Authors and Affiliations

  1. Division of Gastroenterology, University of Arkansas for Medical Sciences and Central Arkansas Veterans Healthcare System, Little Rock, AR, USA

    Julia J. Liu & Elisabeth M. Davis

  2. Wyss Institute for Biologically Inspired Engineering, Harvard University, Boston, MA, USA

    Theresa M. Kay

  3. Division of Gastroenterology, Department of Medicine, University of Alberta, Edmonton, AB, Canada

    Yuefei Lou, Dina Kao & Richard N. Fedorak

  4. Department of Medicine, Brigham and Women’s Hospital, Boston, MA, USA

    Brian Claggett

  5. Department of Medical Microbiology and Immunology, University of Alberta, Edmonton, AB, Canada

    Randall T. Irvin

  6. Division of Gastroenterology and Hepatology, University of Arkansas for Medical Sciences, Shorey S8/62, Little Rock, AR, 72205, USA

    Julia J. Liu

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  1. Julia J. Liu
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  2. Theresa M. Kay
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Correspondence to Julia J. Liu.

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Liu, J.J., Kay, T.M., Davis, E.M. et al. Epithelial Cell Extrusion Zones Observed on Confocal Laser Endomicroscopy Correlates with Immunohistochemical Staining of Mucosal Biopsy Samples. Dig Dis Sci 61, 1895–1902 (2016). https://doi.org/10.1007/s10620-016-4154-x

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  • DOI: https://doi.org/10.1007/s10620-016-4154-x

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