High-throughput rat immunoglobulin G N- glycosylation profiling revealed subclass-specific changes associated with chronic stress (CROSBI ID 296832)
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Habazin, Siniša ; Mlinarević, Dražen ; Balog, Marta ; Bardak, Ana ; Gaspar, Robert ; Szűcs, Kálmán Ferenc ; Vari, Sandor G. ; Vučković, Frano ; Lauc, Gordan ; Novokmet, Mislav ; Heffer, Marija
engleski
High-throughput rat immunoglobulin G N- glycosylation profiling revealed subclass-specific changes associated with chronic stress
Immunoglobulin G (IgG) glycosylation corresponds well with immune system changes, so it can potentially be used as a biomarker for the consequences of chronic stress such as low-grade inflammation and enhanced immunosenescence in older animals. Here we present a high-throughput glycoproteomic workflow, including IgG enrichment, HILIC glycopeptide purification, and nano-LC-MS analysis of tryptic glycopeptides applied for the analysis of rat IgG. A cohort of 80 animals was exposed to seven stressors in a customized chronic stress protocol with blood and tissue sampling in three timepoints. Young female rats experienced an increase in agalactosylated glycoforms on IgG2a and IgG2c accompanied by a decrease in monogalactosylation. Among old females, increased galactosylation was observed in the IgG2b subclass, pointing to an anti-inflammatory activity of IgG. Additionally, IgG Fc N- glycosylation patterns in Sprague Dawley rats were analyzed, quantified, and reported for the first time. Our findings emphasize age-, sex- and subclass-dependent differences in IgG glycosylation related to chronic stress exposure, confirming the relevance of newly developed methods for further research in glycobiology of rodent immune response.
N-glycans ; Glycoproteomics ; Chronic stress ; HPA axis
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