In vivo APC delivery.

To deliver Ag/APC (BMDC or activated B cells), APC were pulsed with 10μM OVA_323–339 (OVA_II) peptide (New England Peptide) or no peptide as a negative control (unpulsed APC) for 1 hour at 37^oC with shaking. APC were washed and administered either intravenously (i.v.) in 200uL PBS, intranasally (i.n.) in 50uL PBS, or intrasplenically (i.s.) in 10uL PBS. 0.25–1×10⁶ BMDC or 1×10⁶ B cells were transferred i.v., 0.5–2×10⁶ BMDC or 1–2×10⁶ B cells were transferred i.n. and 0.5–1×10⁶ B cells were transferred i.s.

Intrasplenic transfer of APC was performed as described previously ¹⁷.