| Phenotypic and Functional Differentiation of Promyelocytic Cell Line HL-60 by N-N-dimethylformamide |
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Kyung Hyo Kim1, Ju Young Seoh2 |
1Department of Pediatrics, College of Medicine, Ewha Womans University, Seoul, Korea
2Department of Midrobiology, College of Medicine, Ewha Womans University, Seoul, Korea |
| N-N-dimethylformamide에 의한 HL-60 세포주의 표현형 및 기능 변화에 관한 연구 |
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김경효1, 서주영2 |
1이화여자대학교 의과대학 소아과학교실
2이화여자대학교 의과대학 미생물학교실 |
Correspondence:
Kyung Hyo Kim, Email: 1
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| Abstract |
Purpose
: HL-60 is a promyelocytic cell line. Fcγ receptors and complement receptor 3(CR3) play
important role in the protective response of granulocytes and monocytes against microbial infection.
We quantified the expression of FcγI, FcγII, FcγIII, and CD11b/CD18 during differentiation using
HL-60 cells by N-N-dimethylformamide(DMF). Functional studies, such as phagocytic activity,
respiratory burst and ADCC, were also performed.
Methods
: HL-60 cells were induced to differentiate by adding 0.8% DMF. On the 4th and 7th day
after stimulation as well as before stimulation, the cells were analyzed for phenotypic and functional
differentiaton. Phenotypic analysis was performed by flow cytometry after staining the cells with
PE-conjugated anti-human CD64, CD32, CD16, CD11b, CD18, and isotype controls. And the measured
fluorescent intensity was transformed into Molecules of Equivalent Soluble Fluorochromes
(MESF). Phagocytic activity was also measured by flow cytometry after incubation with fluorochrome-
conjugated beads. Respiratory burst was measured by chemiluminescence assay of cells incubated
with luminol after stimulation with PMA. ADCC was measured by hemoglobin release
assay.
Results
: The expression of CD11b, CD18 and CD64 on HL-60 cells markedly increased on the 4th
day and slightly decreased on the 7th day. Expression of CD32 was already induced before
differentiation induction and slightly increased by DMF. CD16 was not expressed during differentiation.
In phagocytic assay, the phagocytic cell fraction increased by stimulation on 4th and 7th
day. Chemiluminescence showed the DMF increased the respiratory burst of HL-60 cells on the 4th
and 7th day. In ADCC, DMF increased the target cell lysis continuously.
Conclusion
: HL-60 cells which were differentiated with DMF for are good models for studying
opsonophagocytic assay of immunized sera. |
| Key Words:
HL-60, N-N-dimethylformamide, FcγR, CR3, Function |
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