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. 2020 Apr 27;130(6):2872–2887. doi: 10.1172/JCI131842

Figure 4. Cloned plasma PLA2G1B induces the Bumpy T cell phenotype.

Figure 4

(A) Crystal structure of PLA2G1B (PDB: 6Q42). (B) PLA2G1B effect on MMD formation followed by STED (representative of 2 experiments at 250 nM and verified at 500 nM and 1 μM). (C) Dose effect of PLA2G1B on IL-7–induced p-STAT5 NT in HD purified CD4+ T cells after analysis of confocal images. Results are shown as the mean ± SD from 4 donors. (D–G) The effects on aMMD formation (D and E) and p-STAT5 NT (F and G) in CD4+ T cells of 250 nM WT PLA2G1B were compared to those of the inactive mutant H48Q (D and F) and other PLA2s (PLA2GIIA, PLA2GIID, or PLA2GX) (E and G). Results are shown as the mean ± SD from 5 (D–F) or 7 donors (G). (H) VP plasma (3%, from 5 donors) was depleted with anti-PLA2G1B, anti-PLA2GIIA, or anti-PLA2GIID rabbit polyclonal antibodies (100 μg/mL). The effect of depletion was analyzed by following p-STAT5 NT in IL-7–stimulated CD4+ T cells (n = 3 donors) incubated with depleted plasma. Results were normalized to the response obtained with HD plasma and are shown as the mean ± SD. (I) Effect of VP plasma treated with various doses of neutralizing anti-PLA2G1B mAb 14G9 on p-STAT5 NT in CD4+ T cells from 1 donor and the effect of 100 μg/mL of 14G9 mAb on p-STAT5 NT in CD4+ T cells from 5 donors. **P < 0.01; ***P < 0.001 by Mann-Whitney t test (D, F, and I) and by the Kruskal-Wallis test followed by the Mann-Whitney test with P values adjusted for multiple comparisons between groups (E and G) or 1-way ANOVA (H) with Tukey’s correction for multiple comparisons.