ISS increases the number of mature DCs in FL-treated animals and prolongs the survival of mature DC in vitro.

(A,B) Groups of 3 mice were treated with 9 daily injections of FL followed by one SC injection of ISS (30 μg), C-ODN (30μg), or PBS (FL alone). Forty-eight hours after ISS injection, popliteal and inguinal lymph node cells were isolated and analyzed by flow cytometry. In panel A the contour plots represent IA-b versus CD86 profiles of CD11c⁺ gated cells. The percentages of CD11c⁺cells that are IA-b⁻CD86⁻ and IA-b⁺CD86⁺ are provided in each quadrant. In panel B the number of cells coexpressing IA-b and CD11c was determined on days 1, 7, 12, and 18 after treatment. ■ indicates PBS; ▪, FL-ISS; ▴ FL–C-ODN; ▵, FL alone. (C) The number of IA-b⁺ CD11c⁺ cells coexpressing CD86, CD54, or LFA-1 was determined on day 7 after treatment. These results are the mean of 4 different experiments. ■ indicates PBS; ▪, FL-ISS; ▨, FL–C-ODN; ░, FL alone. (D) BM-DCs were cultured for 11 days in the presence or absence of 10 μg/mL ISS (●), C-ODN (▴), or medium alone (♦). Viable DCs were counted using trypan blue exclusion. Results shown are the mean of 3 separate experiments.