Generation of Aspergillus- and CMV- Specific T Cell Responses Using Autologous Fast-Dendritic Cells (Fast-DC). Session Type: Oral Session

Gamal Ramadan, Steve Konings, Viswanath P. Kurup, Carolyn A. Keever-Taylor

Author address: 

Blood and Marrow Transplantation Program, Medical College of Wisconsin, Milwaukee, WI, USA; Allergy Immunology Division, Veterans Administration Medical Center, Milwaukee, WI, USA


Recent reports have described a new strategy for differentiation and maturation of monocyte-derived dendritic cells within only 48 h of in vitro culture (Fast-DC). In the present study we tested the efficacy of fast-DC to process and present A. fumigatus extract, CMV antigen preparations and CMV-pp65 to autologous T cells compared to DC generated in standard 7-day cultures (standard-DC). Adherent blood monocytes were treated with GM-CSF and IL-4 (1 day for Fast DC, 5 days in the standard arm) to generate immature DC and were then matured for either 1-2 days (Fast DC) or 2 days (standard arm) with IL-1, IL-6, PGE2 and TNF-. The DC were pulsed with Aspergillus or CMV antigen preparations immediately prior to maturation. DC were infected after maturation with an adenovirus vector containing CMV-pp65 (adeno-pp65). Mature DC were then used to prime antigen specific proliferative and cytotoxic T lymphocyte (CTL) responses. Adeno-pp65-infected and Ag-pulsed mature Fast-DC were CD14-, expressed mature DC surface markers (CD40, CD45, CD83, CD86 and HLA-DRbright) and were phenotypically and morphologically indistinguishable from standard-DC. Fast-DC and standard-DC were equally capable of inducing A. fumigatus and CMV-specific T cell proliferation as well priming antigen specific CTL activity. The resulting Aspergillus- and CMV-specific CTL were of mixed CD3+/CD4+ and CD3+/CD8+ phenotype and specifically killed autologous DCs pulsed with A. fumigatus extract and autologous CMV-infected fibroblasts, respectively. These findings indicate that Fast-DC are as effective as standard-DC in the generation of antigen specific T cell responses. Moreover, use of Fast-DC not only reduces labor and supply cost, workload and time but also increases the number of DC derived from adherent monocytes, which may facilitate the use of DCs in clinical trials of cellular immunotherapy. Abstract #134 appears in Blood, Volume 102, issue 11, November 16, 20

abstract No: 


Full conference title: 

American Society of Hematology 45th Annual Meeting
    • ASH 45th (2003)