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. 2014 Jun 2;211(6):1231–1242. doi: 10.1084/jem.20131853

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© 2014 Ali et al.

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Figure 3. — The Siglec-5/14 genotype influences human neutrophil responses to GBS. (A) Human neutrophils of the indicated genotypes were left uninfected or infected with GBS or GBSΔβ (MOI = 5), and bacterial survival at 40 min was assessed by CFU enumeration. (B) Sig-14^+/+ neutrophils were pretreated with the indicated concentrations of anti–Siglec-14 antibody (Ab) for 15 min, followed by GBS infection as in A; after 40 min, bacterial survival was assayed by CFU enumeration. Data are representative of three independent experiments with one donor in each experiment. (C) Human neutrophils of the indicated genotypes were infected with FITC-labeled GBS or with GBSΔβ at 4°C, and then bacterial binding to neutrophils at the 20-min time point was analyzed by FACS. The graph depicts MFI of GFP fluorescence on neutrophils. (D–F) Human neutrophils of the indicated genotypes were left unstimulated or stimulated with LPS, and IL-8 (D), Siglec-5 (E), and Siglec-14 (F) mRNA were analyzed by Q-RT-PCR at 2 h. Results were normalized to the amount of GAPDH mRNA. Data in this figure are representative of two to four independent experiments with one to three different donors per genotype in each experiment. Results are means ± SD; *, P < 0.05, **, P < 0.01.