title: Phenotypic and functional analysis of HL-60 cells used in opsonophagocytic-killing assay for Streptococcus pneumoniae.
abstract: Differentiated HL-60 is an effector cell widely used for the opsonophagocytic-killing assay (OPKA) to measure efficacy of pneumococcal vaccines. We investigated the correlation between phenotypic expression of immunoreceptors and phagocytic ability of HL-60 cells differentiated with N,N-dimethylformamide (DMF), all-trans retinoic acid (ATRA), or 1α, 25-dihydroxyvitamin D3 (VitD3) for 5 days. Phenotypic change was examined by flow cytometry with specific antibodies to CD11c, CD14, CD18, CD32, and CD64. Apoptosis was determined by flow cytometry using 7-aminoactinomycin D. Function was evaluated by a standard OPKA against serotype 19F and chemiluminescence-based respiratory burst assay. The expression of CD11c and CD14 gradually increased upon exposure to all three agents, while CD14 expression increased abruptly after VitD3. The expression of CD18, CD32, and CD64 increased during differentiation with all three agents. Apoptosis remained less than 10% until day 3 but increased after differentiation by DMF or ATRA. Differentiation with ATRA or VitD3 increased the respiratory burst after day 4. DMF differentiation showed a high OPKA titer at day 1 which sustained thereafter while ATRA or VitD3-differentiated cells gradually increased. Pearson analysis between the phenotypic changes and OPKA titers suggests that CD11c might be a useful differentiation marker for HL-60 cells for use in pneumococcal OPKA.
authors: Department of Pediatrics, School of Medicine, Ewha Womans University, Seoul, Korea.
authors: Kyung-Hyo
authors: 0000-0002-0333-6808
authors: KH
authors: Kim
authors: Department of Microbiology, School of Medicine, Ewha Womans University, Seoul, Korea.
authors: Ju Young
authors: 0000-0002-1168-5354
authors: JY
authors: Seoh
authors: Department of Pediatrics, School of Medicine, Ewha Womans University, Seoul, Korea.
authors: Su Jin
authors: 0000-0002-3851-9073
authors: SJ
authors: Cho
mesh_terms: D000907:Antibodies, Bacterial / Q000276:immunology; D017209:Apoptosis / Q000276:immunology*; D001681:Biological Assay; D039521:CD11c Antigen / Q000378:metabolism; D018821:CD18 Antigens / Q000378:metabolism; D002454:Cell Differentiation; D045744:Cell Line, Tumor; D002762:Cholecalciferol / Q000494:pharmacology; D004126:Dimethylformamide / Q000494:pharmacology; D005434:Flow Cytometry; D018922:HL-60 Cells; D006801:Humans; D018950:Lipopolysaccharide Receptors / Q000378:metabolism; D010587:Phagocytosis / Q000276:immunology*; D022242:Pneumococcal Vaccines / Q000276:immunology*; D017452:Receptors, IgG / Q000378:metabolism; D011971:Receptors, Immunologic / Q000096:biosynthesis*; D016897:Respiratory Burst / Q000276:immunology; D013296:Streptococcus pneumoniae / Q000276:immunology*; D014212:Tretinoin / Q000494:pharmacology
chemical_list: D000907:Antibodies, Bacterial; D039521:CD11c Antigen; D018821:CD18 Antigens; D018950:Lipopolysaccharide Receptors; D022242:Pneumococcal Vaccines; D017452:Receptors, IgG; D011971:Receptors, Immunologic; D002762:Cholecalciferol; D014212:Tretinoin; D004126:Dimethylformamide
keywords: Differentiation; HL-60; Opsonophagocytic-Killing Assay; Streptococcus pneumonia
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