Development of a Multiplex Glycan Microarray Assay and Comparative Analysis of Human Serum Anti-Glycan IgA, IgG, and IgM Repertoires

Abstract

Serum antibodies that recognize carbohydrate antigens play a fundamental role in immune defense, homeostasis, and autoimmunity. In addition, they serve as potential biomarkers for a variety of medical applications. For most anti-glycan antibodies found in human serum, however, the origins, regulation, and biological significance are not well understood. Antibody subpopulations that are relevant to a particular biological process or disease are often difficult to identify from the myriad of anti-glycan antibodies present in human serum. While prior studies have examined anti-glycan IgG and/or IgM repertoires, little is known about IgA repertoires or how IgA, IgG, and IgM are related. In this study, we describe the development of a multiplex assay to simultaneously detect IgA, IgG, and IgM on a glycan microarray and its application to studying anti-glycan repertoires in healthy subjects. The multiplex glycan microarray assay revealed unique insights and systems-level relationships that would be difficult to uncover using traditional approaches. In particular, we found that anti-glycan IgA, IgG, and IgM expression levels appear to be tightly regulated, coordinated within individuals, and stable over time. Additionally, our results help define natural fluctuations over time, which is critical for identifying changes that are beyond normal biological variation.

Unique repertoire of anti-carbohydrate antibodies in individual human serum

“…Obviously, this is not the case in natural settings; glycoconjugates in which sugars are linked to proteins or lipids are highly immunogenic and IgG can be directly generated to glycan antigens in that context in rabbits, mice, and humans 18 . In addition, prior studies using glycan microarray technology also supported the possibility that human sera contain a wide range of anti-carbohydrate antibodies of IgG nature 11 , 17 , 19 , 21 , 50 – 52 , 69 .…”

Anti-Glycan Autoantibodies Induced by Helicobacter pylori as a Potential Risk Factor for Myocardial Infarction

“…Glycans decorate the surfaces of all prokaryotic and eukaryotic cells and viruses. Anti-glycan antibodies constitute a large fraction of whole human serum immunoglobulins (26). The best known anti-glycan antibodies are those directed against blood group A and B alloantigens in human plasma.…”

Blood Type Diets (BTD) and Aging: An Overview

“…Because of the various specificities between lectins and the myriad different glycans (interactions between proteins and glycolipid complexes glycolipids and glycolipids, and glycolipids and other carbohydrate complexes), it is not astonishing that the role of food glycans has been missed until now [135,144]. The dearth of information concerning glycans and their presence in food is due essentially to the lack of access to glycans, the poor throughput of traditional assays and the challenges of profiling of AGA [125]. Studies in this field are poised to accelerate greatly due to the availability of high throughput and high-content technologies such as the multiplex glycan bead array (MGBA) [137].…”

“…Dysbiosis will negatively affect health. MOA 4a: Although there is a myriad of AGA present in human serum, their generation and function are not well understood [125]. Normally, healthy humans harbor sets of AGA to blood group-, xeno-(heterophil), and infection-related glycotopes [160].…”

“…(b) Variation of the composition of the gut microbiota (positively or negatively], which will consequently influence inflammation[124]. Be absorbed into the internal milieu by diffusion or pinocytosis where they (a) Are presented to the APCs (antigen-presenting cells), such as dendritic cells, which will present the glycan for the generation of specific anti-glycan antibodies (AGA)[34,125]. (b) Interact with the complement proteins through the lectin pathway.…”