Figure 4
Production of bacteria-reactive antibodies in mouse peritoneal cavity cells following treatment with CpG-DNA 1826 in vitro. BALB/c mice were i.p. injected with PBS, CpG-DNA 1826, or non-CpG-DNA 2041. (A) After 7 days, the cells from the peritoneal cavity were harvested and then stimulated with PBS, CpG-DNA 1826, or non-CpG-DNA 2041. After 48 h, the cell culture supernatants were collected, and the amounts of total IgG were determined by ELISA (n = 3/group). (B) The peritoneal cells from PBS-injected mice were stimulated with PBS, CpG-DNA 1826, or non-CpG-DNA 2041. To measure the amounts of antibodies reactive to Gram-positive bacteria, the indicated bacteria were added to poly-L-lysine-coated plates and the cell culture supernatants were applied. The amount of bound IgG was determined by ELISA (n = 3/group). (C) After the i.p. administration of PBS or CpG-DNA 1826 to BALB/c mice, B1 and B2 cells were isolated from the peritoneal cavity using a FACSAriaTM II system and fluorescence-labeled anti-mouse CD19 and anti-mouse CD23 antibodies. (D,E) Isolated B1 and B2 cells from the peritoneal cavity were stimulated with PBS or CpG-DNA 1826. After 48 h, the cell culture supernatants were collected and the amount of total IgG (D) and the antibodies reactive to Gram-positive bacteria (E) were determined by ELISA (n = 3/group). 1826, CpG-DNA 1826. 2041, non-CpG-DNA 2041. The results presented are representative of three experiments. ***p < 0.0005.
