Fast generation of dendritic cells

TY - JOUR

T1 - Fast generation of dendritic cells

AU - Kvistborg, P

AU - Bøgh, Marie

AU - Pedersen, A W

AU - Claesson, M H

AU - Pedersen, A W

N1 - Keywords: Antigens, CD; Cell Culture Techniques; Cell Differentiation; Coculture Techniques; Dendritic Cells; Flow Cytometry; Humans; Lymphocyte Culture Test, Mixed; Phenotype; T-Lymphocytes; Time; Vaccines

PY - 2009

Y1 - 2009

N2 - Dendritic cells (DC) are potent antigen presenting cells capable of inducing immune responses. DC are widely used as vaccine adjuvant in experimental clinical settings. DC-based vaccines are normally generated using a standard 8day DC protocol (SDDC). In attempts to shorten the vaccine production we have developed fast DC protocol by comparing two different fast DC protocols with SDDC. DC were evaluated by FACS analysis, and the optimal profile was considered: CD14(low), CD80(high), CD83(high), CD86(high), CCR7(high), HLA class I and II(high). FACS profiles were used as the selection criteria together with yield and morphology. Two fast DC protocols fulfilled these criteria and were selected for functional analysis. Our results demonstrate that DC generated within 5days or 48h are comparable with SDDC both phenotypically and functionally. However, we found that 48h DC were more susceptible than SDDC to the IL-10 inducing stimulus of TLR ligands (R848 and LPS). Thus to determine the clinical relevance of fast DC protocols in cancer settings, small phase I trials should be conducted monitoring regulatory T cells carefully.

AB - Dendritic cells (DC) are potent antigen presenting cells capable of inducing immune responses. DC are widely used as vaccine adjuvant in experimental clinical settings. DC-based vaccines are normally generated using a standard 8day DC protocol (SDDC). In attempts to shorten the vaccine production we have developed fast DC protocol by comparing two different fast DC protocols with SDDC. DC were evaluated by FACS analysis, and the optimal profile was considered: CD14(low), CD80(high), CD83(high), CD86(high), CCR7(high), HLA class I and II(high). FACS profiles were used as the selection criteria together with yield and morphology. Two fast DC protocols fulfilled these criteria and were selected for functional analysis. Our results demonstrate that DC generated within 5days or 48h are comparable with SDDC both phenotypically and functionally. However, we found that 48h DC were more susceptible than SDDC to the IL-10 inducing stimulus of TLR ligands (R848 and LPS). Thus to determine the clinical relevance of fast DC protocols in cancer settings, small phase I trials should be conducted monitoring regulatory T cells carefully.

U2 - 10.1016/j.cellimm.2009.09.003

DO - 10.1016/j.cellimm.2009.09.003

M3 - Journal article

C2 - 19818956

SN - 0008-8749

VL - 260

SP - 56

EP - 62

JO - Cellular Immunology

JF - Cellular Immunology

IS - 1

ER -