- Top of page
- Generation of Df-specific, MoDCs
- Flowcytometric analysis
- Cell preparation
- Ag-specific T-cell responses
- Cytokine production assays and determination
- Statistical analysis
- Effect of Df prepulse on the expression of surface molecules on MoDCs
- Roles of Df-prepulsed MoDCs in the activation of Ag-specific memory T cells in the absence of recall stimulation
- Effect of Df-prepulsed MoDCs on cytokine production by Ag-specific memory T cells
- Duration of Ag presentation by Df-prepulsed MoDCs
- Role of Df-prepulsed MoDCs in the activation of naïve T cells in the absence of recall stimulation
Background: Dendritic cells are one of the most potent antigen-presenting cells and when pulsed with allergen can modulate allergen-specific T-cell responses. We sought to establish a single-step method by which to generate allergen-specific monocyte-derived dendritic cells (MoDCs).
Methods: Dermatophagoides farinae (Df)-prepulsed MoDCs were generated from monocytes by culturing with Df in the presence interleukin (IL)-4, granulocyte-macrophage colony-stimulating factor (GM-CSF) and tumour necrosis factor (TNF)-α simultaneously. Df-prepulsed MoDC were incubated with autologous naïve and memory T cells in the absence of recall antigen, then proliferation and cytokine production by T cells was determined.
Results: Generation of allergen-prepulsed MoDCs was confirmed by examining expression of surface molecules. Df-prepulsed MoDC selectively induced proliferation of Df-specific T cells in the absence of recall antigen. Under these conditions, Df-prepulsed MoDCs augmented but did not alter the cytokine production profile. In addition, Df-prepulsed MoDCs activated naïve T cells leading to proliferation and selective production of IFN-γ in allergic patients but not in healthy subjects.
Conclusions: These results suggest that Df-prepulsed MoDC generated from monocytes by a simple single-step manipulation can induce Df-specific cellular responses from both naïve and memory T cells in the absence of recall antigen, and these cells potentially can be utilized as immune adjuvants in allergen-specific immunotherapy.