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Phenotypic and functional analysis of HL-60 cells used in opsonophagocytic-killing assay for Streptococcus pneumoniae

Title
Phenotypic and functional analysis of HL-60 cells used in opsonophagocytic-killing assay for Streptococcus pneumoniae
Authors
Kim K.-H.Seoh J.Y.Cho S.J.
Ewha Authors
서주영김경효조수진
SCOPUS Author ID
서주영scopus; 김경효scopus; 조수진scopus
Issue Date
2015
Journal Title
Journal of Korean Medical Science
ISSN
1011-8934JCR Link
Citation
vol. 30, no. 2, pp. 145 - 150
Keywords
DifferentiationHL-60Opsonophagocytic-killing assayStreptococcus pneumonia
Publisher
Korean Academy of Medical Science
Indexed
SCI; SCIE; SCOPUS; KCI WOS scopus
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 1a, 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 ATRAor 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. © 2015 The Korean Academy of Medical Sciences.
DOI
10.3346/jkms.2015.30.2.145
Appears in Collections:
의학전문대학원 > 의학과 > Journal papers
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