Application of enzyme-linked immunosorbent assay to detect antimicrobial peptides in human intestinal lumen.

Autor: Hong JS; Columbia Center of Translational Immunology, Department of Medicine, Columbia University, New York, NY, United States of America. Electronic address: julie.hong2@gmail.com., Shamim A; Columbia Center of Translational Immunology, Department of Medicine, Columbia University, New York, NY, United States of America; College of Dental Medicine, Columbia University, New York, NY, United States of America., Atta H; Columbia Center of Translational Immunology, Department of Medicine, Columbia University, New York, NY, United States of America., Nonnecke EB; Department of Microbiology and Immunology, University of California Davis School of Medicine, Davis, CA, United States of America., Merl S; Department of Pathology and Cell Biology, Columbia University, New York, NY, United States of America., Patwardhan S; Columbia Center of Translational Immunology, Department of Medicine, Columbia University, New York, NY, United States of America., Manell E; Columbia Center of Translational Immunology, Department of Medicine, Columbia University, New York, NY, United States of America; Department of Clinical Sciences, Swedish University of Agricultural Sciences, Uppsala, Sweden., Gunes E; Columbia Center of Translational Immunology, Department of Medicine, Columbia University, New York, NY, United States of America., Jordache P; Columbia Center of Translational Immunology, Department of Medicine, Columbia University, New York, NY, United States of America., Chen B; Columbia Center of Translational Immunology, Department of Medicine, Columbia University, New York, NY, United States of America., Lu W; Department of Biochemistry and Molecular Biology, University of Maryland School of Medicine, Baltimore, MD, United States of America., Shen B; Department of Surgery, Columbia University/New York-Presbyterian Hospital, New York, NY, United States of America., Dionigi B; Department of Surgery, Columbia University/New York-Presbyterian Hospital, New York, NY, United States of America., Kiran RP; Department of Surgery, Columbia University/New York-Presbyterian Hospital, New York, NY, United States of America., Sykes M; Columbia Center of Translational Immunology, Department of Medicine, Columbia University, New York, NY, United States of America; Department of Surgery, Columbia University/New York-Presbyterian Hospital, New York, NY, United States of America., Zorn E; Columbia Center of Translational Immunology, Department of Medicine, Columbia University, New York, NY, United States of America., Bevins CL; Department of Microbiology and Immunology, University of California Davis School of Medicine, Davis, CA, United States of America., Weiner J; Columbia Center of Translational Immunology, Department of Medicine, Columbia University, New York, NY, United States of America; Department of Surgery, Columbia University/New York-Presbyterian Hospital, New York, NY, United States of America.
Jazyk: angličtina
Zdroj: Journal of immunological methods [J Immunol Methods] 2024 Feb; Vol. 525, pp. 113599. Date of Electronic Publication: 2023 Dec 09.
DOI: 10.1016/j.jim.2023.113599
Abstrakt: Intestinal transplantation is the definitive treatment for intestinal failure. However, tissue rejection and graft-versus-host disease are relatively common complications, necessitating aggressive immunosuppression that can itself pose further complications. Tracking intraluminal markers in ileal effluent from standard ileostomies may present a noninvasive and sensitive way to detect developing pathology within the intestinal graft. This would be an improvement compared to current assessments, which are limited by poor sensitivity and specificity, contributing to under or over-immunosuppression, respectively, and by the need for invasive biopsies. Herein, we report an approach to reproducibly analyze ileal fluid obtained through stoma sampling for antimicrobial peptide/protein concentrations, reasoning that these molecules may provide an assessment of intestinal homeostasis and levels of intestinal inflammation over time. Concentrations of lysozyme (LYZ), myeloperoxidase (MPO), calprotectin (S100A8/A9) and β-defensin 2 (DEFB2) were assessed using adaptations of commercially available enzyme-linked immunosorbent assays (ELISAs). The concentration of α-defensin 5 (DEFA5) was assessed using a newly developed sandwich ELISA. Our data support that with proper preparation of ileal effluent specimens, precise and replicable determination of antimicrobial peptide/protein concentrations can be achieved for each of these target molecules via ELISA. This approach may prove to be reliable as a clinically useful assessment of intestinal homeostasis over time for patients with ileostomies.
Competing Interests: Declaration of Competing Interest None.
(Copyright © 2023. Published by Elsevier B.V.)
Databáze: MEDLINE
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