Toxoplasmacidal effect of HL-60 cells differentiated by dimethylsulfoxide

, Choi, Won Young; , Nam, Ho Woo; , Yoo, Jae Eul

doi:1988.26.4.229

Korean J Parasito > Volume 26(4):1988 > Article

Original Article


Korean J Parasitol. 1988 Dec;26(4):229-238. English. Published online Mar 20, 1994. http://dx.doi.org/10.3347/kjp.1988.26.4.229
Copyright © 1988 by The Korean Society for Parasitology


Toxoplasmacidal effect of HL-60 cells differentiated by dimethylsulfoxide
Won Young Choi,Ho Woo Nam and Jae Eul Yoo
Department of Parasitology, Catholic University Medical College, Seaol 135-040, Korea.


Abstract
In vitro culture of Toxoplasma gondii in HL-60 cells cnd cell-mediated immunity against Toxoplasma in dimethylsulfoxide(DMSO)-induced HL-60 cells, i.e., differentiation into granulocytes, were pursued. HL-60 cells were treated with various concentrations of DMSO, and 1.3%(v/v) for 3 day incubation was chosen as the optimal condition for differentiation into granulocytes. The degree of differentiation was assayed in physiological and functional aspects in addition to morphological point. When treated with 1.3% DMSO for 3 days, HL-60 cells did not synthesize DNA materials beyond background level, and showed active chemotactic response to chemotactic peptide, formyl-methionyl-leucyl-phenylalanine(FMLP). Morphologically promyelocytes of high nuclear/cytoplasmic(N/C) ratio changed to granulocytes of relatively low N/C ratio. The relationships between HL-60 cells or DMSO-induced HL-60 cells and Toxoplasma were examined after stain with Giemsa and fluorescent dye (acridine orange). HS-60 cells did not show any sign of toxoplasmacidal activity but showed intracellular proliferation of Toxoplasma to form rosette for 72 hr co-culture. In contrast, DMSO-induced HL-60 cells phagocytosed Toxoplasma within 1 hr, and performed a process of intracellular digestion of Toxoplasma thereafter. With the above results, it is suggested that phagosome-lysosome fusion is one of the critical events for the parasitism by Toxoplasma or for susceptibility of host cells. The in vitro culture system of this study has offered a defined condition to study the protozoan parasite-host cell interactions.

Figures


	Fig. 1 Dose-response growth of HL-60 cells in DMSO. Cells were inoculated at 3×10⁵ cells/ml in the presence of various concentrations of DMSO.


	Fig. 2 ³H-thymidine incorporation by HL-60 cells in the presence of 1.3% DMSO. Untreated or DMSO-treated cells were labeled pulsely with ³H-thymidine for 1 hr, then the percentages were calculated.


	Fig. 3 Chemotactic response of HL-60 cells or DMSO-induced HL-60 cells to FMLP. Percentages of the cells migrated through the filter to initially inoculated were calculated.

Fig. 4
Giemsa-stained photographs of co-culture of HL-60 cells (HC, H1, H2, H3, and H4) or DMSO-induced HL-60 cells (DC, D1, D2, D3, and D4) with Toxoplasma. HC and DC stand for the controls of HL-60 cells and DMSO-induced HL-60 cells. H1 and D1 represent the co-culture of HL-60 cells or DMSO-induced HL-60 cells with Toxoplasma after 1 hr; H2 and D2, 5 hr; H3 and D3, 24 hr; and H4 and D4, 72 hr; respectively.

Fig. 5
Fluorescent photographs of co-culture of HL-60 cells (HFC, HF1, HF2, HF3, and HF4) or DMSO-induced HL-60 cells (DFC, DF1, DF2, DF3, and DF4) with Toxoplasma, HFC and DFC stand for the controls of HL-60 cells or DMSO-induced HL-60 cells. HF1 and DF1 represent the co-culture of HL-60 cells or DMSO-induced HL-60 cells after 1 hr; HF2 and DF2, 5 hrs; HF3 and DF3, 24 hrs; and HF4 and DF4, 72 hrs; respectively.

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