title: Monitoring and isolation of blood dendritic cells from apheresis products in healthy individuals: a platform for cancer immunotherapy.
abstract: The fundamental role of dendritic cells (DC) in initiating and directing the primary immune response is well established. Furthermore, it is now accepted that DC may be useful in new vaccination strategies for preventing certain malignant and infectious diseases. As blood DC (BDC) physiology differs from that of the DC homologues generated in vitro from monocyte precursors, it is becoming more relevant to consider BDC for therapeutic interventions. Until recently, protocols for the isolation of BDC were laborious and inefficient; therefore, their use for investigative cancer immunotherapy is not widespread. In this study, we carefully documented BDC counts, yields and subsets during apheresis (Cobe Spectra), the initial and essential procedure in creating a BDC isolation platform for cancer immunotherapy. We established that an automated software package (Version 6.0 AutoPBPC) provides an operator-independent reliable source of mononuclear cells (MNC) for BDC preparation. Further, we observed that BDC might be recovered in high yields, often greater than 100% relative to the number of circulating BDC predicted by blood volume. An average of 66 million (range, 17-179) BDC per 10-l procedure were obtained, largely satisfying the needs for immunization. Higher yields were possible on total processed blood volumes of 15 l. BDC were not activated by the isolation procedure and, more importantly, both BDC subsets (CD11c(+)CD123(low) and CD11c(-)CD123(high)) were equally represented. Finally, we established that the apheresis product could be used for antibody-based BDC immunoselection and demonstrated that fully functional BDC can be obtained by this procedure.
authors.affiliation: Mater Medical Research Institute, Aubigny Place, South Brisbane 4101, Australia
authors.forename: J
authors.initials: J
authors.lastname: Alejandro López
authors.forename: Georgina
authors.initials: G
authors.lastname: Crosbie
authors.forename: Cathryn
authors.initials: C
authors.lastname: Kelly
authors.forename: Ann Marie
authors.initials: AM
authors.lastname: McGee
authors.forename: Katrina
authors.initials: K
authors.lastname: Williams
authors.forename: Slavica
authors.initials: S
authors.lastname: Vuckovic
authors.forename: Robert
authors.initials: R
authors.lastname: Schuyler
authors.forename: Robyn
authors.initials: R
authors.lastname: Rodwell
authors.forename: Sue J
authors.initials: SJ
authors.lastname: Wright
authors.forename: Kerry
authors.initials: K
authors.lastname: Taylor
authors.forename: Derek N J
authors.initials: DN
authors.lastname: Hart
mesh_terms: D000328:Adult; D001781:Blood Component Removal; D002469:Cell Separation / Q000379:methods*; D003713:Dendritic Cells / Q000166:cytology* / Q000276:immunology*; D005260:Female; D005434:Flow Cytometry; D006801:Humans; D007167:Immunotherapy / Q000379:methods*; D007963:Leukocytes, Mononuclear / Q000166:cytology / Q000276:immunology; D007959:Lymphocyte Culture Test, Mixed; D008297:Male; D008875:Middle Aged; D009369:Neoplasms / Q000276:immunology / Q000628:therapy*