title: Day-4 myeloid dendritic cells pulsed with whole tumor lysate are highly immunogenic and elicit potent anti-tumor responses.
abstract: "Day-7" myeloid DCs are commonly used in the clinic. However, there is a strong need to develop DCs faster that have the same potent immunostimulatory capacity as "Day-7" myeloid DCs and at the same time minimizing time, labor and cost of DC preparations. Although "2 days" DCs can elicit peptide-specific responses, they have not been demonstrated to engulf, process and present complex whole tumor lysates, which could be more convenient and personalized source of tumor antigens than defined peptides. In this preclinical study, we evaluated the T-cell stimulatory capacity of Day-2, Day-4, and Day-7 cultured monocyte-derived DCs loaded with SKOV3 cell whole lysate prepared by freeze-thaw or by UVB-irradiation followed by freeze-thaw, and matured with lipopolysaccharide (LPS) and interferon (IFN)-gamma. DCs were evaluated for antigen uptake, and following maturation with LPS and IFN-gamma, DCs were assessed for expression of CD80, CD40, CD86, ICAM-1 and CCR7, production of IL-12p70 and IP-10, and induction of tumor-specific T-cell responses. Day-4 and Day-7 DCs exhibited similar phagocytic abilities, which were superior to Day-2 DCs. Mature Day-7 DCs expressed the highest CD40 and ICAM-1, but mature Day-4 DCs produced the most IL-12p70 and IP-10. Importantly, Day-4 and Day-7 DCs derived from ovarian cancer patients stimulated equally strongly tumor-specific T-cell responses. This is the first study demonstrating the highly immunogenic and strong T-cell stimulatory properties of Day-4 myeloid DCs, and provided important preclinical data for rapid development of potent whole tumor lysate-loaded DC vaccines that are applicable to many tumor types.
authors.affiliation: Ovarian Cancer Research Center, University of Pennsylvania, Philadelphia, Pennsylvania, United States of America.
authors.forename: Cheryl Lai-Lai
authors.initials: CL
authors.lastname: Chiang
authors.forename: Andrea R
authors.initials: AR
authors.lastname: Hagemann
authors.forename: Rachel
authors.initials: R
authors.lastname: Leskowitz
authors.forename: Rosemarie
authors.initials: R
authors.lastname: Mick
authors.forename: Thomas
authors.initials: T
authors.lastname: Garrabrant
authors.forename: Brian J
authors.initials: BJ
authors.lastname: Czerniecki
authors.forename: Lana E
authors.initials: LE
authors.lastname: Kandalaft
authors.forename: Daniel J
authors.initials: DJ
authors.lastname: Powell
authors.forename: George
authors.initials: G
authors.lastname: Coukos
mesh_terms: D002454:Cell Differentiation / Q000187:drug effects; D045744:Cell Line, Tumor; D045142:Cross-Priming / Q000187:drug effects; D016207:Cytokines / Q000096:biosynthesis; D003713:Dendritic Cells / Q000166:cytology / Q000187:drug effects / Q000276:immunology*; D005260:Female; D006262:Health; D006801:Humans; D007371:Interferon-gamma / Q000494:pharmacology; D008070:Lipopolysaccharides / Q000494:pharmacology; D022423:Myeloid Cells / Q000166:cytology / Q000187:drug effects / Q000276:immunology*; D010051:Ovarian Neoplasms / Q000276:immunology* / Q000473:pathology*; D010587:Phagocytosis / Q000187:drug effects; D010641:Phenotype; D013601:T-Lymphocytes / Q000166:cytology / Q000187:drug effects / Q000276:immunology; D014019:Tissue Donors; D014020:Tissue Extracts / Q000494:pharmacology*
chemical_list: D016207:Cytokines; D008070:Lipopolysaccharides; D014020:Tissue Extracts; D007371:Interferon-gamma