Neutrophils are front-line immune cells critical for the innate immune system. Many aspects of the structure and function of the neutrophil glycoproteome remain unresolved. Enabled by technology advancements in glycomics and glycoproteomics, the intriguing protein N-glycosylation of human neutrophils was investigated in the context of cancer, inflammation and infection. A novel class of truncated N-glycoproteins, paucimannosidic proteins, was discovered in sputum from pathogen-infected human lungs (Venkatakrishnan et al., Glycobiology, 25(1):88, 2015). The monosaccharide compositions i.e. Man1-3GlcNAc2Fuc0-1, the biosynthetic machinery involving maturation stage-specific β-hexosaminidase expression and the preferential location in azurophilic granules of neutrophils were demonstrated (Thaysen-Andersen et al., J Biol Chem, 290(14):8789, 2015). Importantly, these compartment- and inflammation-associated glyco-signatures were carried by intact bioactive proteins including cathepsin G, azurocidin and neutrophil elastase (Loke et al., Biomolecules, 5(3):1832, 2015). Glycomics-based studies on isolated neutrophil granules confirmed that paucimannosylation is enriched on azurophilic granules-resident proteins, but present also in other compartments. Paucimannosidic proteins were preferentially secreted, but not incorporated into the plasma membrane above constitutive levels, upon inoculation of isolated neutrophils with virulent P. aeruginosa thereby confirming granular mobility of paucimannosidic proteins. Preliminary data shows that isolated paucimannosidic proteins displayed affinities to mannose-binding lectin and bacteriostatic activities towards virulent P. aeruginosa supporting immune-related functions of paucimannosylation in activated human neutrophils. Interestingly, isolated neutrophils from Sandhoff disease patients displaying a HEXB-/- genotype showed reduced protein paucimannosylation relative to age-paired healthy donors confirming the importance of β-hexosaminidases in paucimannosidic protein biosynthesis. Finally, other immune and cancer cells were shown to also express paucimannosidic glycans indicating that these under-reported glycoproteins are integral to the immune system across cell types. In conclusion, we provide insights into the intriguing features of neutrophil glycobiology by expanding our knowledge of the structure, function and biosynthesis of the spatiotemporally-regulated protein paucimannosylation in the context of cancer, inflammation and infection.