Zebrafish mount a Csf3r-dependent granulopoietic response to T. marneffei infection.
(A) Low magnification images of infected and uninfected whole Tg(mpx:EGFP) embryos at 0, 3 and 4 dpi, displaying the profound infection-driven increase in EGFP-expressing neutrophil numbers. (B, C) Neutrophil (B) and macrophage (C) population expansion in infected versus uninfected embryos from 0–4 dpi. These data derive from enumerating neutrophil and macrophage numbers in the same embryos, of genotype Tg(mpx:EGFP/mpeg1:Gal4FF/UAS-E1b:Eco.nfsB-mCherry). Data displayed are mean±SD, n = 4 embryos/group. These data replicate other data derived from separately scoring these leukocyte populations in different embryos, in n≥2 independent other experiments. (D) Neutrophil population sizes at 4 dpi represented by Neutrophil Units in embryos injected with antisense morpholino oligonucleotides targeted to knockdown csf3-receptor (csf3r-MO) or interleukin-6 receptor (il6ra-MO and gp130-MO) Data are mean±SEM, n = 10 embryos/group/experiment, n≥3 experiments. Intragroup +/- infection p-values from 2-tailed unpaired t-test; p-values from intergroup comparisons involving csf3r-MO from 1-tailed unpaired t-test, given the a priori expectation of a reduction in neutrophil values in csf3r-deficiency [65, 95] (E) Kaplan-Meier life table analysis for the treatment groups shown in (C). Numbers/group (uninfected/infected): Control MO, 474/326; csf3r-MO, 443/233; il6ra-MO, 462/330; gp130-MO, 465/313. P-values from Gerhan-Breslow-Wilcoxon Test.