Neurol Neuroimmunol Neuroinflamm 2016 Dec 28;3(6):e284. Epub 2016 Sep 28.
Institute of Infection, Immunity and Inflammation (S.K.H., H.J.W.), College of Medical, Veterinary and Life Sciences, University of Glasgow; The Beatson Institute for Cancer Research (G.K.), Glasgow, UK; Laboratory Sciences and Services Division (M.B.I., I.J., Q.D.M., Z.I.), International Centre for Diarrheal Disease Research; Shaheed Tajuddin Ahmed Sarani (M.B.I., I.J., Q.D.M., Z.I.), Mohakhali, Dhaka, Bangladesh; Departments of Immunology and Neurology (B.C.J.) and Medical Microbiology and Infectious Diseases (H.P.E.), Erasmus MC, University Medical Center Rotterdam, Rotterdam, the Netherlands; and Fondation Mérieux (H.P.E.), Lyon, France.
Objective: To characterize the patterns of autoantibodies to glycolipid complexes in a large cohort of Guillain-Barré syndrome (GBS) and control samples collected in Bangladesh using a newly developed microarray technique.
Methods: Twelve commonly studied glycolipids and lipids, plus their 66 possible heteromeric complexes, totaling 78 antigens, were applied to polyvinylidene fluoride-coated slides using a microarray printer. Arrays were probed with 266 GBS and 579 control sera (2 μL per serum, diluted 1/50) and bound immunoglobulin G detected with secondary antibody. Scanned arrays were subjected to statistical analyses.
Results: Measuring antibodies to single targets was 9% less sensitive than to heteromeric complex targets (49.2% vs 58.3%) without significantly affecting specificity (83.9%-85.0%). The optimal screening protocol for GBS sera comprised a panel of 10 glycolipids (4 single glycolipids GM1, GA1, GD1a, GQ1b, and their 6 heteromeric complexes), resulting in an overall assay sensitivity of 64.3% and specificity of 77.1%. Notable heteromeric targets were GM1:GD1a, GM1:GQ1b, and GA1:GD1a, in which exclusive binding to the complex was observed.
Conclusions: Rationalizing the screening protocol to capture the enormous diversity of glycolipid complexes can be achieved by miniaturizing the screening platform to a microarray platform, and applying simple bioinformatics to determine optimal sensitivity and specificity of the targets. Glycolipid complexes are an important category of glycolipid antigens in autoimmune neuropathy cases that require specific analytical and bioinformatics methods for optimal detection.