Strains of bacterial species induce a greatly varied acute adaptive immune response: The contribution of the accessory genome
A. CFSE labeled PBMC from 10 donors were stimulated and re-stimulated as described in Fig 1A (as in Fig 1) with either Newman or USA600, stained and analyzed by FACS for percent CFSE dilution (%proliferation) in live CD3+CD4+ cells that were gated on Th subsets based on cell surface markers (CxCR3, CCR6, CCR4). B. Example of FACS plot of one of the donors in Figure A. C. Absolute number of proliferating cells in response to stimulation with either Newman or USA600, when gated on cell surface markers for Th subsets cells. D. Intracellular staining for IFNγ, IL17A expression in proliferating live CD3+CD4+ cells following stimulation with either Newman or USA600 as in A. E. As in D, but staining for transcription factor Tbet and RORγt. F. As in D after stimulation with Streptococcus pyogenes strains. G. The MANOVA test for difference between bivariate means of induced IFNγ+IL17A- vs IL17A+IFNγ- absolute (Abs) number of cells following stimulation with Streptococcus pyogenes strains among 10 donors, showed significant results (p<0.00001). The post-hoc pairwise comparisons identified the differences between strains to be significant (for M1 vs M3 and M1 vs M6, P<0.0001; for M3 vs M6 P<0.005).