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. 2016 Jun 24;11(6):e0158019. doi: 10.1371/journal.pone.0158019

Fig 5. Bacterial uptake and survival following co-incubation with neutrophils isolated from hyperglycemic and normoglycemic mice.

Fig 5

Bone marrow (A-B and E-F) or peritoneally-recruited (C-D and G-J) neutrophils were isolated from B. burgdorferi-infected and mock-infected C57BL/6 (A, C, E, G) and C3H/HeN (B, D, F, H-J) mice at 4 weeks post-infection (5–6 weeks of sustained hyperglycemia), and coincubated with complement-opsonized E. coli (A-D) or B. burgdorferi (E-J). E. coli survival (A-D) and B. burgdorferi uptake by neutrophils (E-H) were measured by comparing numbers of E. coli CFUs and intact B. burgdorferi following neutrophil co-incubation with values for complement-opsonized input bacteria. (I-J) B. burgdorferi killing was measured by LIVE-DEAD staining. Panel I shows sample images for bacteria incubated in the absence (no PMN) and presence (+PMN) of neutrophils. Panel J shows quantification of B. burgdorferi killing. Summary values: mean ±SEM. N = 4–6 mice/group. Experimental groups: No neutrophil (PMN) control: opsonized bacteria incubated in absence of neutrophils, normoglycemic mock-infected (Vh), hyperglycemic mock-infected (STZ), normoglycemic B. burgdorferi-infected (Vh+Bb), hyperglycemic B. burgdorferi-infected (STZ+Bb). Statistical analysis: Two-way ANOVA with Holm-Sidak post-tests. * indicates p<0.05 vs. Vehicle; # indicates p<0.05 vs. normoglycemic B. burgdorferi-infected (Vh+Bb) mice; † indicates p<0.05 vs. hyperglycemic mock-infected mice (STZ); & indicates p<0.05 vs no PMN control (panel J only).