Fig 6. RTX treatment reduces the frequencies of CD11b⁺ Ly6G⁺ICAM-1⁺ myeloid cells in the infected corneas.

A. Histogram plots show gating and frequencies of CD11b⁺ Ly6G⁺ICAM-1^- and CD11b⁺Ly6G⁺ ICAM-1⁺ cells. Data represent 6 infected corneas (1 per mouse) per condition per cohort. The experiment was repeated twice. Bar graphs show mean values; symbols represent individual mouse. P-values by Student’s t-test. B. Representative ImageStream images of CD11b⁺ Ly6G⁺ICAM-1⁺ and CD11b⁺ Ly6G⁺ ICAM-1^- cells. Mice were infected with 5×10⁵ CFU of GFP-expressing P. aeruginosa 6294 (Green) per cornea; tissue was harvested at 24h post-challenge for analysis. Cellular suspensions were prepared with collagenase digestion and stained for CD45, CD11b, Ly6G, ICAM-1, and DNA (Hoechst) to identify myeloid infiltrates. CD11b⁺ Ly6G⁺ cells show banded or multi-lobed nuclear morphology typical of neutrophils. C. Quantification of CD11b⁺ Ly6G⁺ICAM-1⁺ and CD11b⁺ Ly6G⁺ ICAM-1^- myeloid cells containing intracellular P. aeruginosa-GFP (p-values by one-way ANOVA). Cumulatively, data show that nociceptor presence alters myeloid phenotypes in the infected corneas, enriching for CD11b⁺ ICAM-1⁺ Ly6G⁺ myeloid cells in vehicle-treated mice when compared to RTX-treated mice.