Abstract
This report describes an experimental procedure for constructing integrated lipid, carbohydrate, and protein microarrays. In essence, it prints liposomes on nitrocellulose-coated micro-glass slides, a biochip substrate for spotting protein and carbohydrate microarrays, and the substances that can form liposomes (homo-liposomes) or can be incorporated into liposomes (hetero-liposomes) are suitable for microarray construction using existing microarray spotting devices. Importantly, this technology allows simultaneous detection of serum antibody activities among the three major classes of antigens, i.e., lipids, carbohydrates, and proteins. The potential of this technology is illustrated by its use in revealing a broad-spectrum of pre-existing anti-lipid antibodies in blood circulation and monitoring the epitope spreading of autoantibody reactivities among protein, carbohydrate, and lipid antigens in experimental autoimmune encephalomyelitis (EAE).
Highlights
Like proteins and carbohydrates, lipids are a category of the essential elements of living cells
A key question for this liposome array technology is whether the spotted liposomes preserve the antigenic determinants that are readily reactive with specific anti-lipid antibodies
If the liposome arrays produced by this procedure preserve the lipid epitopes that are readily reactive with anti-lipid antibodies and if the assay reaches the sensitivity to detect these antibodies in blood circulation, using these arrays to scan the serum specimens collected from either normal mice or EAE subjects would allow detection of corresponding anti-lipid antibodies in these models
Summary
Lipids are a category of the essential elements of living cells. Microarrays 2015, 4 microbes, such as lipid A components of LPS, which are ligands of the Toll-like receptors of the innate immune system [5,6] Host recognition of such lipids leads to rapid first-line anti-infection responses. Campylobacter jejuni infection induced an autoimmune neurological disorder, Guillain-Barré syndrome, in about a third of cases [7,8] This pathogen expresses a lipopolysaccharide molecule that mimics various gangliosides present in high concentrations in peripheral nerves. Immunization with cell-wall polysaccharide of Streptococcus pneumoniae elicited T15 anti-phosphorylcholine antibodies, which cross-react with oxidized epitopes of low-density lipoprotein (oxLDL) This antibody response was found to be effective in eliminating oxLDL in circulation and in atherosclerotic lesions [17,19]. A practical experimental procedure was explored to enable high-throughput production of lipid microarrays using this biochip substrate
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