All issues > Volume 41(10); 1998

Original Article

J Korean Pediatr Soc. 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 HS Kim¹, Kyung Hyo KH Kim¹, Gyoung Hee GH Kim¹, Ju Young JY Seoh²

¹Department of Pediatrics, College of Medicine, Ewha Womans University, Seoul, Korea
²Department of Microbiology, College of Medicine, Ewha Womans University, Seoul, Korea

Correspondence Hye Soon HS 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.

Keywords :HL-60 cells, FcγRⅠ, FcγRⅡ, FcγRⅢ, CD11b/CD18