Dendritic cells (DCs) are considered to be the most efficient antigen presenting cells. Intratracheal administration of allergen-pulsed bone marrow-derived dendritic cells (BMDCs) prior to allergen challenge induces airway hyperresponsiveness (AHR) and inflammation. Ovalbumin (OVA)-pulsed BMDCs from wild-type (WT) mice were transferred into naive WT, CD4-/-, CD8-/-, or IL-13-/- mice. Two (short protocol) or 10 days (long protocol) after BMDC transfer, mice were challenged with 1% OVA for 3 days and assayed 2 days later. Transfer of OVA-primed BMDCs into BALB/c or C57BL/6 mice elicited AHR in both protocols. Airway eosinophilia, Th2 cytokines, or goblet cell metaplasia were only increased in the long but not short protocol. Lung T cells from both protocols produced Th2 cytokines in response to OVA in vitro. Carboxyfluorescein diacetate succinimidylester (CFSE)-labeled BMDCs were observed in bronchoalveolar (BAL) fluid and lung parenchyma at early time points, and were detected in draining lymph nodes 48 hrs after transfer. CD8-/- mice developed AHR comparable to WT mice in the short protocol, but decreased levels of AHR, airway eosinophilia, Th2 cytokines in BAL fluid, and goblet cell metaplasia compared to WT mice in the long protocol. CD4-/- or IL-13-/- mice did not develop AHR or airway inflammation in either protocol. These data suggest that allergen-pulsed BMDCs initiate development of AHR which is dependent initially on CD4+ T cells, and at later time periods on CD8+ T cells and IL-13. Thus, the timing of allergen challenge following transfer of allergen-pulsed BMDC affects the development of AHR and airway inflammation.