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Immunologic Changes of HL-60 Cells by Differentiation Inducing Agents I : Phenotypic Differentiation of FcγR and Mac-1

Journal of the Korean Pediatric Society 1998;41(10):1342-1350.
Published online October 15, 1998.
Immunologic Changes of HL-60 Cells by Differentiation Inducing Agents I : Phenotypic Differentiation of FcγR and Mac-1
Hye Soon Kim1, Kyung Hyo Kim1, Gyoung Hee Kim1, Ju Young Seoh2
1Department of Pediatrics, College of Medicine, Ewha Womans University, Seoul, Korea
2Department of Microbiology, College of Medicine, Ewha Womans University, Seoul, Korea
HL-60 세포주의 분화유도제 처리에 따른 면역학적 변화 Ⅰ: FcγR와 Mac-1 표현형에 관한 연구
김혜순1, 김경효1, 김경희1, 서주영2
1이화여자대학교 의과대학 소아과학교실
2이화여자대학교 의과대학 미생물학교실
Correspondence: 
Hye Soon Kim, Email: 1
Abstract
Purpose
: Fc receptors and Mac-1 play an important role in the protective response of granulocytes and monocytes against microbial infection. FcγRⅠ, FcγRⅡ, FcγRⅢ as well as CD11b/ CD18 have never been measured in a quantitative way during hemopoiesis. Thus we quantified the expression of FcγRⅠ, FcγRⅡ, FcγRⅢ, and CD11b/CD18 during hematopoietic differentiation using HL-60 cells, which was induced to differentiate by DMSO, or PMA.
Methods
: HL-60 cells(ATCC CCL-240) were induced to differentiate by adding 1.0% DMSO, or 16nM PMA. On the 4th and 7th day after stimulation as well as before stimulation, 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).
Results
: Percent positive cells and MESF of CD11b on HL-60 cells increased upon induction by DMSO, but not by PMA. Percent positive cells of CD18 on HL-60 cells was 99% regardless of differentiation. But MESF of CD18 was increased on the 4th day and decreased on the 7th day by DMSO or PMA. Percent positive cells and MESF of FcγRⅠ on HL-60 cells increased upon induction by DMSO or PMA. Percent positive cells of FcγRⅡ on HL-60 cells was above 90% regardless of differentiation. MESF of FcγRⅡ showed no significant change by DMSO or PMA.
Conclusion
: Quantitative expression of FcγRⅠ, FcγRⅡ, FcγRⅢ, and CD11b/CD18 of HL-60 cells changed during induction of differentiation by DMSO or PMA. MESF of FcγR and CD11b/ CD18 a better indicator than percent positive cells to compare the differentiation of HL-60cells.
Key Words: HL-60 cells, FcγRⅠ, FcγRⅡ, FcγRⅢ, CD11b/CD18


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