|   | 
Details
   web
Record
Author (up) Schnabel, C.L.; Babasyan, S.; Freer, H.; Wagner, B.
Title Quantification of equine immunoglobulin A in serum and secretions by a fluorescent bead-based assay Type Journal Article
Year 2017 Publication Veterinary Immunology and Immunopathology Abbreviated Journal
Volume 188 Issue Pages 12-20
Keywords Horse; Immunoglobulin A; Monoclonal antibody; Fluorescent bead-based assay; Mucosal secretion
Abstract Abstract Only few quantitative reports exist about the concentrations and induction of immunoglobulin A (IgA) in mucosal secretions of horses. Despite this, it is widely assumed that IgA is the predominant immunoglobulin on mucosal surfaces in the horse. Here, two new monoclonal antibodies (mAbs) against equine IgA, clones 84-1 and 161-1, were developed and characterized in detail. Both IgA mAbs specifically bound monomeric and dimeric equine IgA in different applications, such as Western blots and fluorescent bead-based assays. Cross-reactivity with other equine immunoglobulin isotypes was not observed. The new IgA mAb 84-1 was used in combination with the previously characterized anti-equine IgA mAb BVS2 for the development and validation of a fluorescent bead-based assay to quantify total IgA in equine serum and various secretions. The IgA assay's linear detection ranged from 64 pg/ml to 1000 ng/ml. For the quantification of IgA in serum or in secretions an IgA standard was purified from serum or nasal wash fluid (secretory IgA), respectively. The different standards were needed for accurate IgA quantification in the respective samples taking the different signal intensities of monomeric and dimeric IgA on the florescent bead-based assay into account. IgA was quantified by the bead-based assay established here in different equine samples of healthy adult individuals. In serum the median total IgA was 0.45 mg/ml for Thoroughbred horses (TB, n = 10) and 1.16 mg/ml in Icelandic horses (ICH, n = 12). In nasopharyngeal secretions of TB (n = 7) 0.13 mg/ml median total IgA was measured, and 0.25 mg/ml for ICH (n = 12). Saliva of ICH (n = 6) contained a median of 0.15 mg/ml, colostrum of Warmbloods (n = 8) a median of 1.89 mg/ml IgA. Compared to IgG1 and IgG4/7 quantified in the same samples, IgA appeared as the major immunoglobulin isotype in nasopharyngeal secretions and saliva while it is a minor isotype in serum and colostrum. The newly developed monoclonal antibodies against equine IgA and the resulting bead-based assay for quantification of total IgA can notably improve the evaluation of mucosal immunity in horses.
Address
Corporate Author Thesis
Publisher Place of Publication Editor
Language Summary Language Original Title
Series Editor Series Title Abbreviated Series Title
Series Volume Series Issue Edition
ISSN 0165-2427 ISBN Medium
Area Expedition Conference
Notes Approved no
Call Number Equine Behaviour @ team @ Serial 6152
Permanent link to this record