Profiling and genetic control of the murine immunoglobulin G glycome

Abstract

Immunoglobulin G (IgG) glycosylation is essential for function of the immune system, but the genetic and environmental factors that underlie its inter-individual variability are not well defined. The Collaborative Cross (CC) genetic resource harnesses over 90% of the common genetic variation of the mouse. By analyzing the IgG glycome composition of 95 CC strains, we made several important observations: (i) glycome variation between mouse strains was higher than between individual humans, despite all mice having the same environmental influences; (ii) five genetic loci were found to be associated with murine IgG glycosylation; (iii) variants outside traditional glycosylation site motifs affected glycome variation; (iv) bisecting N-acetylglucosamine (GlcNAc) was produced by several strains although most previous studies have reported the absence of glycans containing the bisecting GlcNAc on murine IgGs; and (v) common laboratory mouse strains are not optimal animal models for studying effects of glycosylation on IgG function.

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Fig. 1: Representative overlaid chromatograms of mouse and human IgG glycans separated by UPLC.
Fig. 2: Variation in the levels of individual IgG glycans between different strains of mice.
Fig. 3: Sex-dependent differences in the levels of individual IgG glycans.
Fig. 4: LC-MS glycopeptide data plotted versus UPLC glycan data.
Fig. 5: QTL mapping of glycosylation traits.

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Acknowledgements

We thank Geniad Pty Ltd for generously providing CC mice and genotypes. This project was supported in part by the WA Diabetes Research Foundation (to G.M.), Program 1037321 (to G.M.) and Project 1069173 (to G.M.) from the National Health and Medical Research Council of Australia and by European Commission FP7 grants MIMOmics (contract 305280 to G.L.) and H2020 projects GlySign (contract 722095 to G.L.), SYSCID (contract #733100 to G.L.) and IMforFuture (contract 721815 to G.L.), as well as by the European Structural and Investment Funds IRI (grant KK.01.2.1.01.0003 to G.L.) and Croatian National Centre of Research Excellence in Personalized Healthcare (grant KK.01.1.1.01.0010 to G.L.).

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Q.N., K.M.D. and G.M. provided samples; J.K., O.O.Z., I.T.-A., M.N., M.P., G.M. and G.L. planned experiments; J.K., M.N., M.V. and O.O.Z. performed experiments; R.R., O.O.Z. and F.V. analyzed data; J.K., G.M. and G.L. wrote the manuscript.

Corresponding author

Correspondence to Gordan Lauc.

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Competing interests

G.L. is founder and owner of Genos Ltd, which specializes in high-throughput glycomics analysis and has several patents in the field. G.M. holds stock in Geniad Pty Ltd.

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Supplementary Text and Figures

Supplementary Figures 1–9 and Supplementary Tables 1–8

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Supplementary Dataset 1

List of candidate genes involved in the regulation of mouse IgG glycosylation identified from the QTL analysis.

Supplementary Dataset 2

Strain, sex and age of mice.

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Krištić, J., Zaytseva, O.O., Ram, R. et al. Profiling and genetic control of the murine immunoglobulin G glycome. Nat Chem Biol 14, 516–524 (2018). https://doi.org/10.1038/s41589-018-0034-3

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