High prevalence of subclass-specific binding and neutralizing antibodies against Clostridium difficile toxins in adult cystic fibrosis sera: possible mode of immunoprotection against symptomatic C. difficile infection

Abstract

Tanya M Monaghan,1 Ola H Negm,2,3 Brendon MacKenzie,4 Mohamed R Hamed,2,3 Clifford C Shone,5 David P Humphreys,4 K Ravi Acharya,6 Mark H Wilcox7 1Nottingham Digestive Diseases Centre, NIHR Nottingham Digestive Diseases Biomedical Research Unit, School of Medicine, University of Nottingham, Nottingham, 2Breast Surgery Group, Division of Medical Sciences and Graduate Entry Medicine, School of Medicine, Queen’s Medical Centre, University of Nottingham, Nottingham, UK; 3Medical Microbiology and Immunology Department, Faculty of Medicine, Mansoura University, Mansoura, Egypt; 4Antibody Biology, UCB-New Medicines, UCB Celltech, Slough, UK; 5Toxins Group, National Infection Service, Public Health England, Salisbury, UK; 6Department of Biology and Biochemistry, University of Bath, Bath, UK; 7Leeds Institute of Biomedical and Clinical Sciences, University of Leeds, Leeds, UK Objectives: Despite multiple risk factors and a high rate of colonization for Clostridium difficile, the occurrence of C. difficile infection in patients with cystic fibrosis is rare. The aim of this study was to compare the prevalence of binding C. difficile toxin-specific immunoglobulin (Ig)A, IgG and anti-toxin neutralizing antibodies in the sera of adults with cystic fibrosis, symptomatic C. difficile infection (without cystic fibrosis) and healthy controls. Methods: Subclass-specific IgA and IgG responses to highly purified whole C. difficile toxins A and B (toxinotype 0, strain VPI 10463, ribotype 087), toxin B from a C. difficile toxin-B-only expressing strain (CCUG 20309) and precursor form of B fragment of binary toxin, pCDTb, were determined by protein microarray. Neutralizing antibodies to C. difficile toxins A and B were evaluated using a Caco-2 cell-based neutralization assay. Results: Serum IgA anti-toxin A and B levels and neutralizing antibodies against toxin A were significantly higher in adult cystic fibrosis patients (n=16) compared with healthy controls (n=17) and patients with symptomatic C. difficile infection (n=16); p≤0.05. The same pattern of response prevailed for IgG, except that there was no difference in anti-toxin A IgG levels between the groups. Compared with healthy controls (toxins A and B) and patients with C. difficile infection (toxin A), sera from cystic fibrosis patients exhibited significantly stronger protective anti-toxin neutralizing antibody responses.Conclusion: A superior ability to generate robust humoral immunity to C. difficile toxins in the cystic fibrosis population is likely to confer protection against symptomatic C. difficile infection. This protection may be lost in the post-transplantation setting, where sera monitoring of anti-C. difficile toxin antibody titers may be of clinical value. Keywords: Clostridium difficile, cystic fibrosis, antibodies  

Keywords

• Clostridium difficile Cystic fibrosis Antibodies