Strains of bacterial species induce a greatly varied acute adaptive immune response: The contribution of the accessory genome
A. A donor CFSE labeled PBMC were stimulated with 16 heat killed S. aureus strains and 4 days later were re-stimulated for 6 hrs after which they were stained and analyzed by FACS for percent CFSE dilution (proliferation) and IFNγ expression in proliferating cells when gated on live CD3+CD4+ cells. B. Example of FACS plot of 2 of 16 strains evaluated using same Figure A donor. C. Upper panel is the percent live CD3+CD4+ proliferation in 10 donors in response to 16 strains. The pair-wise comparison is in S1 Table. Lower panel is live CD3+CD4+ proliferation (lower left panel), and IFNγ expression in live CD3+CD4+ proliferating cells (lower right panel) in same 10 donors in response to 8 representative strains (with high, intermediate and low T cell proliferation). The results from all 16 strains for percent IFNγ expression and the complete statistical analysis are presented in the S1 Fig and S2 Table. For lower panels, Friedman statistics P<0.0001 for both, significant Dunns for pairs in Rt lower panel: for Newman vs 8325, Newman vs USA600, NRS111 vs 8325, NRS111 vs USA600, USA500 vs USA700, USA500 vs 8325, USA500 vs USA600. In left lower panel: Newman vs 8325, Newman vs USA600, NRS111 vs USA700, NRS111 vs 8325, NRS111 vs USA100, NRS111 vs USA600, USA500 vs 8325, USA500 vs USA600. D. Same donor as in A, but gated on live CD3-CD19+ for B cell proliferation and IgG expression in proliferating cells in response to the 16 strains. E. Example of FACS plot of 2 of 16 strains evaluated in Figure D donor. F. The percent B cell proliferation and IgG expression in live CD3-CD19+ proliferating cells in 10 donors in response to the 8 strains with prominent different values C. (the results from the 16 strains are in Supporting Information). G. The percent proliferation and IFNγ expression by live CD3+CD4+ proliferating cells stimulated with in to Streptococcus pyogenes strains in 10 donors. H. and I. The MANOVA test for the difference between bivariate means of ranks of T and B cells proliferation (H) and IFNγ vs IgG expression (I) among 16 strains, showed significant differences (p<0.00001). The post-hoc pairwise comparisons identified the differences between strains to be significant (P<0.0001), except for T and B cell proliferation (H) with Mu50 vs. USA 100 (p-value = 0.53), and IFNγ vs IgG expression (I) with Newman vs USA100 (p-value = 0.51).