COAM induces peritoneal alterations in leukocyte populations in animals with hyperacute EAE. Peritoneal cells were collected on days 0, 5, 9 and 14, 15 or 16 post immunization with SCH in SJL/J mice. (A) The percentages (morphologic examinations) of peritoneal macrophages and (B) neutrophils in the peritoneal exudates were compared between naive mice and EAE-induced mice, treated i.p. with saline or with COAM. Data are compiled from three experiments; bars represent averages ± SEM of three to nine mice. Statistical significance was determined using the Mann Whitney test. (C-E) Flow cytometric analysis of leukocytes in the peritoneal cavity of COAM-treated versus saline-treated EAE mice. Cells were stained for the presence of CD11b, Gr-1, CD4 and CD8. (C) Example of FACS analysis with indication of gating of double positive cells. The numbers within the gates indicate the percentages of Gr-1-positive cells present in the whole sorted peritoneal lavages. One representative analysis out of nine is shown. (D-E) The data of three experiments were pooled and the percentages of CD4+ and CD8+ cells on each of the examined days are shown. Bars represent averages ± SEM, the individual numbers of animals (n) for each condition are indicated below each of the histograms. Asterisks indicate * P <0.05; ** P <0.01 for comparison with naive mice (Mann Whitney test). Pairwise comparisons (P values) between saline- (control) and COAM-treated hyperacute EAE mice are indicated on top of the histograms. COAM, chlorite-oxidized oxyamylose; EAE, experimental autoimmune encephalomyelitis; SCH, spinal cord homogenate; SEM, standard error of the mean.