A method for in vivo analysis of gastric mucin oligosaccharides was developed and applied to rhesus monkeys with and without Helicobacter pylori infection. Mucin-type O-linked oligosaccharides were directly released by reductive beta-elimination from gastric biopsies from rhesus monkeys. The released oligosaccharides were structurally characterized by matrix-assisted laser desorption/ionozation and electrospray ionization Fourier transform ion cyclotron resonance mass spectrometry. A diverse profile of neutral and acidic oligosaccharides was observed with these techniques. The most predominant core structure detected in all of the samples at relatively high abundance corresponded to core 2 (2HexNAc-1Hex, m/z = 611.227). The spectra generated from H. pylori-infected monkey samples showed fewer oligosaccharides collectively. Peaks corresponding to 1HexNAc-1Hex (m/z = 408.148) and 2HexNAc (m/z = 449.174), which most likely represent core structures, were absent in all infected monkeys studied, although present in all uninfected monkeys. Unsupervised cluster analysis demonstrated clear differences between the peaks detected in uninfected and naturally infected monkey samples. The results suggest that H. pylori infection is associated with lower relative abundance of oligosaccharides and loss of mucin-type core structures. This method can be applied to characterize the glycans associated with the mucin lining of live animals and allows for repeated analysis of the same animal over the course of infection.
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