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. 2020 Sep 23;18:262. doi: 10.1186/s12916-020-01721-8

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© The Author(s) 2020

Open AccessThis article is licensed under a Creative Commons Attribution 4.0 International License, which permits use, sharing, adaptation, distribution and reproduction in any medium or format, as long as you give appropriate credit to the original author(s) and the source, provide a link to the Creative Commons licence, and indicate if changes were made. The images or other third party material in this article are included in the article's Creative Commons licence, unless indicated otherwise in a credit line to the material. If material is not included in the article's Creative Commons licence and your intended use is not permitted by statutory regulation or exceeds the permitted use, you will need to obtain permission directly from the copyright holder. To view a copy of this licence, visit http://creativecommons.org/licenses/by/4.0/. The Creative Commons Public Domain Dedication waiver (http://creativecommons.org/publicdomain/zero/1.0/) applies to the data made available in this article, unless otherwise stated in a credit line to the data.

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Fig. 4 — Glycan microarray analysis shows high anti-Neu5Gc IgG specificity and increased levels and diversity with higher Neu5Gc intake. Human serum IgG (n = 120; 1/100 dilution) detected with Cy3-anti-human IgG by glycan microarrays (24-pairs Neu5Gc-/Neu5Ac-glycans, αGal; Additional file 1: Table S3, Additional file 3: Data file S2). Relative fluorescence units (RFU) normalized to IgG (ng/μl) against printed standard curve/array [30]. a Sum serum IgG response/individual against Neu5Gc-/Neu5Ac-glycans, per Neu5Gc intake quartile, showed specific anti-Neu5Gc IgG (mean ± sem; Friedman ANOVA, p < 0.0001), a trend of elevated anti-Neu5Gc IgG at higher Neu5Gc intake. b Serum IgG/individual against αGal showed no change in levels/Neu5Gc intake quartile. c Men 45–60 stratified according to total Neu5Gc intake (Q1–Q4), contributing dietary sources plotted, showing increased Neu5Gc intake between quartiles, red meat dominant. d Men 45–60, antibodies/quartile show increased anti-Neu5Gc IgG levels between Q1 and Q2–Q4 (sum mean IgG response/glycan across individuals; different colors/specific Neu5Gc-glycan). e Pie charts of sum anti-Neu5Gc IgG response (d) divided per quartile according to reactivity against Neu5Gc-glycans with different Sia-linkages (Siaα2–3/6/8 linkages: α3, α6, α8, respectively) or underlying glycans [Lac (lactose; Galβ3Glc), Gal (galactose), type 1 (Galβ3GlcNAc), GalNAc (N-acetylgalactoseamine), LacNAc (N-acetyllactoseamine; Galβ4GlcNAc), core 1 (Galβ3GalNAcα)]. Differences in diversity at higher Neu5Gc intake, characterized by increased levels of α3-linked-Neu5Gc and Lac underlying glycans. f Women aged 45–60 stratified according to total Neu5Gc intake (Q1–Q4), increase between quartiles, similar contributions of Neu5Gc intake from red meat and dairy cow, dominance for dairy cow. g Women 45–60, anti-Neu5Gc IgG reactivity per quartile increase in levels between Q1 and Q2–Q4. h Women 45–60, pie charts of sum anti-Neu5Gc IgG response (g) divided by characteristic Neu5Gc-glycans linkage/skeleton, differences in diversity, as in men