friedman test with dunn’s multiple comparisons test  (GraphPad Software Inc)

Journal: Frontiers in Immunology

Article Title: A Quantitative Approach to Unravel the Role of Host Genetics in IgG-FcγR Complex Formation After Vaccination

doi: 10.3389/fimmu.2022.820148

Figure Lengend Snippet: IgG1 allotype determines whether boosting IgG1 concentration or boosting IgG1 affinity (k on IgG1- FcγR) would be most effective for increasing complex formation. (A) Model predictions for complex formation of RV144 vaccinees (n=105) in two FcγRIIIa polymorphisms, FcγRIIIa-V 158 (light pink) and FcγRIIIa-F 158 (dark pink), and three IgG1 allotypes, G1m1,3 (original RV144 data), G1m1 and G1m-1,3. Polymorphisms were simulated by altering the binding affinities of each IgG subtype to FcγR as previously published and indicated in Figure 3A . Allotypes are simulated by multiplying each vaccinee’s IgG1, IgG2, IgG3 and IgG4 initial concentration by its respective conversion factor as previously published and indicated in Figure 4A (Friedman test with Dunn’s multiple comparisons test comparing the two polymorphisms within each allotype; ****p-value < 0.001). (B) Simulated IgG1 concentration boosting in each allotype (G1m1,3, white; G1m1, gray; G1m-1,3 black) and polymorphism (FcγRIIIa-V 158 , light pink; FcγRIIIa-F 158 , dark pink) combination. Boosts were calculated by multiplying the individual’s baseline initial IgG1 concentration value by the boost levels and then this was added on top of each individual’s baseline. (B) Color indicates median change in complex formation for each genetic background. (C) Simulated boosting of k on IgG1- FcγR in each allotype (G1m1,3, white; G1m1, gray; G1m-1,3 black) and polymorphism (FcγRIIIa-V 158 , light pink; FcγRIIIa-F 158 , dark pink) combination. Boosts were calculated by multiplying the individual’s baseline k on IgG1- FcγR value by the boost levels and then this was added on top of each individual’s baseline. Color indicates median change in complex formation for each genetic background and boost as indicated. (D) The ratio of median change in complex formation with a boost in IgG1 concentration over median change in complex formation with a boost in k on IgG1-FcγR (affinity) at each boosting level. This ratio shows which type of boost is most effective for increasing complex formation (IgG1 concentration, purple; k on IgG1-FcγR, green) and when both are equally beneficial (white).

Article Snippet: In order to evaluate affinity changes resulting from glycosylation, projected upon all vaccinees for each of the three allotypes, the IgG-FcR immune complex formation was simulated at baseline, and the difference between each individual’s complex formation at baseline and with glycosylation for each allotyped population and compared them with a Friedman test with Dunn’s multiple comparisons in GraphPad Prism.

Techniques: Concentration Assay, Binding Assay

Journal: Frontiers in Immunology

Article Title: A Quantitative Approach to Unravel the Role of Host Genetics in IgG-FcγR Complex Formation After Vaccination

doi: 10.3389/fimmu.2022.820148

Figure Lengend Snippet: Glycosylation differentially impacts IgG1 allotypes. (A) Expected IgG1, IgG2, IgG3, and IgG4 concentrations for G1m1,3 (white), G1m1 (gray), and G1m-1,3 (black) allotypes based on previously published work ( , ). (B) Model predictions for complex formation as IgG1 concentration and k on IgG1- FcγR are altered over physiological ranges ( Figure 2B ). Lines indicate IgG1 concentrations for three different IgG1 allotypes (G1m1,3 (white), G1m1 (gray), G1m-1,3 (black)), and the affinity change expected from an afucosylation glycosylation modification (purple) compared to baseline (light blue). (C) The difference ( Figure 2C ) between the combined parameter change surface ( Figure 2A ) and the additive surface ( Figure 2B ). Lines indicate IgG1 concentrations for three different IgG1 allotypes (G1m1,3 (white), G1m1 (gray), G1m-1,3 (black)), and the affinity change expected from an afucosylation glycosylation modification (dark blue) compared to baseline FcgRIIIaV158 (light blue). (D) Change in complex formation from baseline affinity to an afucosylated affinity in each allotype, G1m1,3 (white), G1m1 (gray), and G1m-1,3 (black) (Friedman test with Dunn’s multiple comparisons test; ****p-value < 0.001).

Article Snippet: In order to evaluate affinity changes resulting from glycosylation, projected upon all vaccinees for each of the three allotypes, the IgG-FcR immune complex formation was simulated at baseline, and the difference between each individual’s complex formation at baseline and with glycosylation for each allotyped population and compared them with a Friedman test with Dunn’s multiple comparisons in GraphPad Prism.

Techniques: Glycoproteomics, Concentration Assay, Modification

Journal: Cell Reports Medicine

Article Title: A systems approach to elucidate personalized mechanistic complexities of antibody-Fc receptor activation post-vaccination

doi: 10.1016/j.xcrm.2021.100386

Figure Lengend Snippet: IgG1 concentration differences resulting from Gm allotype are predicted to significantly alter FcR complex formation (A) Conversion factors for each initial IgG concentration from G1m1,3 to indicated allotypes. Projections were simulated by multiplying each vaccinee’s initial IgG concentrations by the respective conversion factors and performing the simulations according to the baseline protocol. (B) Model predicted complex formation for FcγRIIIa-V 158 in G1m-1,3 (n = 105; black circles) and G1m1 (n = 105; white circles) compared with the original data, assumed to be G1m1,3 (n = 105; red circles) ∗p < 0.05, ∗∗p < 0.01, ∗∗∗p < 0.001, > ∗∗∗p < 0.0001, using the Friedman test with Dunn’s multiple comparisons test.

Article Snippet: : Complex formation for each Gm allotype were compared using a Friedman’s test with Dunn’s multiple comparison test in GraphPad Prism (α = 0.05).

Techniques: Concentration Assay