Profiling Humoral Immune Responses to Clostridium difficile-Specific Antigens by Protein Microarray Analysis

Negm, Ola H.; Hamed, Mohamed R.; Dilnot, Elizabeth M.; Shone, Clifford C.; Marszalowska, Izabela; Lynch, Mark; Loscher, Christine E.; Edwards, Laura J.; Tighe, Patrick J.; Wilcox, Mark H.; Monaghan, Tanya M.

Abstract

Clostridium difficile is an anaerobic, Gram-positive, and spore-forming bacterium that is the leading worldwide infective cause of hospital-acquired and antibiotic-associated diarrhea. Several studies have reported associations between humoral immunity and the clinical course of C. difficile infection (CDI). Host humoral immune responses are determined using conventional enzyme-linked immunosorbent assay (ELISA) techniques. Herein, we report the first use of a novel protein microarray assay to determine systemic IgG antibody responses against a panel of highly purified C. difficile-specific antigens, including native toxins A and B (TcdA and TcdB, respectively), recombinant fragments of toxins A and B (TxA4 and TxB4, respectively), ribotypespecific surface layer proteins (SLPs; 001, 002, 027), and control proteins (tetanus toxoid and Candida albicans). Microarrays were probed with sera from a total of 327 individuals with CDI, cystic fibrosis without diarrhea, and healthy controls. For all antigens, precision profiles demonstrated

• Journal Article Type: Article
• Acceptance Date: Jul 2, 2015
• Online Publication Date: Jul 15, 2015
• Publication Date: 2015-09
• Deposit Date: Dec 7, 2015
• Publicly Available Date: Dec 7, 2015
• Journal: Clinical and Vaccine Immunology
• Print ISSN: 1556-6811
• Electronic ISSN: 1556-679X
• Publisher: American Society for Microbiology
• Peer Reviewed: Peer Reviewed
• Volume: 22
• Issue: 9
• Pages: 1033-1039
• DOI: https://doi.org/10.1128/CVI.00190-15
• Keywords: Clostridium difficile, Antigen Microarray
• Public URL: https://nottingham-repository.worktribe.com/output/982458
• Publisher URL: http://cvi.asm.org/content/22/9/1033.full