Figure 5. Murine acute inflammation.

A. Mouse bone marrow cells (mBMC) and neutrophils (mPMN) were isolated from 10-12 week C57BL/6 mice, and cultured with 50μM HexNAc analogs or controls for 40h. Cells were analyzed using flow cytometry and used in a murine acute inflammation model. B.-E. Flow cytometry measured the binding of the following reagents to mouse neutrophils (CD11b+, Gr-1/Ly-6G/1A8+, F4/80−): B. VVA-FITC, C. Anti-mouse PSGL-1 mAb 2PH1, D. P-selectin-IgG and E. L-selectin-IgG. Ac₅GalNTGc increased VVA-lectin binding by 4-5 fold, and reduced L-/P-selectin IgG binding by 50–70% without affecting PSGL-1 expression. F. mBMCs cultured with 80μM Ac₅GalNTGc for 40h were mixed with VC at 1:1 ratio. In mix 1, Ac₅GalNTGc cells were labeled with CMTMR (Red) while VC was CMFDA (Green) labeled. Labels were swapped in Mix 2 (dot plot not shown). Mix 1 or 2 cells were tailvein injected into recipient mice following thioglycollate injection i.p. Red:green ratio of Gr-1+ cells in the peritoneal lavage and bone marrow was measured at 20h. Ac₅GalNTGc reduced neutrophil counts in peritoneum by 50% in both Mixes. G-I. Ac₅GalNTGc (100mg/kg/day) or VC was injected daily into mice for 4 days prior to induction of peritonitis. Murine neutrophil (CD11b+, Gr-1/Ly-6G/1A8+, F4/80−) counts in the peritoneum were quantified at 16h. Neutrophil counts in peritoneal lavage was reduced by 65% in Ac₅GalNTGc treated mice (panel H). VVA binding was augmented in peritoneal neutrophils (panel I). *P<0.05 with respect to all other treatments in each panel, except as indicated.