Phenotypic and Functional Analysis of HL-60 Cells Used in Opsonophagocytic-Killing Assay for Streptococcus pneumoniae

다국어 초록 (Multilingual 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 correlationbetween phenotypic expression of immunoreceptors and phagocytic ability of HL-60 cellsdifferentiated with N,N-dimethylformamide (DMF), all-trans retinoic acid (ATRA), or 1α,25-dihydroxyvitamin D3 (VitD3) for 5 days. Phenotypic change was examined by flowcytometry with specific antibodies to CD11c, CD14, CD18, CD32, and CD64. Apoptosiswas determined by flow cytometry using 7-aminoactinomycin D. Function was evaluatedby a standard OPKA against serotype 19F and chemiluminescence-based respiratory burstassay. The expression of CD11c and CD14 gradually increased upon exposure to all threeagents, while CD14 expression increased abruptly after VitD3. The expression of CD18,CD32, and CD64 increased during differentiation with all three agents. Apoptosis remainedless 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. DMFdifferentiation showed a high OPKA titer at day 1 which sustained thereafter while ATRAorVitD3-differentiated cells gradually increased. Pearson analysis between the phenotypicchanges and OPKA titers suggests that CD11c might be a useful differentiation marker forHL-60 cells for use in pneumococcal OPKA.

참고문헌 (Reference)

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