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Investigation of Chemotactic Activities in Differentiated HL-60 Cells by a Time-lapse Videomicroscopic Assay

Jung, Yun-Jae   (Department of Microbiology, Ewha Womans University, Laboratory of ImmunodynamicsUU0001056  ); Woo, So-Youn   (Department of Microbiology, Ewha Womans University, Laboratory of ImmunodynamicsUU0001056  ); Ryu, Kyung-Ha   (Department of Pediatrics, College of Medicine, Ewha Womans University, Laboratory of ImmunodynamicsUU0001056  ); Jang, Myoung-Ho   (Department of Microbiology and Immunology, Osaka University Graduate School of MedicineUU0014187  ); Miyasaka, Masayuki   (Department of Microbiology and Immunology, Osaka University Graduate School of MedicineUU0014187  ); Seoh, Ju-Young   (Department of Microbiology, Ewha Womans University, Laboratory of ImmunodynamicsUU0001056  );
  • 초록

    Background: Chemotaxis is one of the cardinal functions of leukocytes, which enables them to be recruited efficiently to the right place at the right time. Analyzing chemotactic activities is important not only for the study on leukocyte migration but also for many other applications including development of new drugs interfering with the chemotactic process. However, there are many technical limitations in the conventional in vitro chemotaxis assays. Here we applied a new optical assay to investigate chemotactic activities induced in differentiated HL-60 cells. Methods: HL-60 cells were stimulated with 0.8% dimethylformamide (DMF) for 4 days. The cells were analyzed for morphology, flow cytometry as well as chemotactic activities by a time-lapse videomicroscopic assay using a chemotactic microchamber bearing a fibronectin-coated cover slip and an etched silicon chip. Results: Videomicroscopic observation of the real cellular motions in a stable concentration gradient of chemokines demonstrated that HL-60 cells showed chemotaxis to inflammatory chemokines (CCL3, CCL5 and CXCL8) and also a homeostatic chemokine (CXCL12) after DFM-induced differentiation to granulocytic cells. The cells moved randomly at a speed of $6.99{\pm}1.24{\mu}m/min$ (n=100) in the absence of chemokine. Chemokine stimulation induced directional migration of differentiated HL-60 cells, while they still wandered very much and significantly increased the moving speeds. Conclusion: The locomotive patterns of DMF-stimulated HL-60 cells can be analyzed in detail throughout the course of chemotaxis by the use of a time-lapse videomicroscopic assay. DMF-stimulated HL-60 cells may provide a convenient in vitro model for chemotactic studies of neutrophils.

  • 주제어

    Chemotaxis .   videomicroscopy .   HL-60 cells .   leukocyte migration .   boyden chamber .   transwell assay.  

  • 참고문헌 (32)

    1. Howe RS, Fischkoff SA, Rossi RM, Lyttle CR: Chemotactic capabilities of HL-60 human myeloid leukemia cells differentiated to eosinophils. Exp Hematol 18;299-303, 1990 
    2. Chaplinski TJ, Sloan GJ, Niedel JE: Granulocyte functions during maturation of human promyelocytic leukemia cells. Leuk Res 9;897-903, 1985 
    3. Matzner Y, Gavison R, Rachmilewitz EA, Fibach E: Expression of granulocytic functions by leukemic promyelocytic HL-60 cells: differential induction by dimethylsulfoxide and retinoic acid. Cell Differ 21;261-269, 1987 
    4. Koeffler HP, Bar-Eli M, Territo MC: Phorbol ester effect on differentiation of human myeloid leukemia cell lines blocked at different stages of maturation. Cancer Res 41;919-926, 1981 
    5. Tanaka T, Ebisuno Y, Kanemitsu N, Umemoto E, Yang BG, Jang MH, Miyasaka M: Molecular determinants controlling homeostatic recirculation and tissue-specific trafficking of lymphocytes. Int Arch Allergy Immunol 134;120-134, 2004 
    6. Boyden S: The chemotactic effect of mixtures of antibody and antigen on polymorphonuclear leucocytes. J Exp Med 115;453-466, 1962 
    7. Zigmond SH: Ability of polymorphonuclear leukocytes to orient in gradients of chemotactic factors. J Cell Biol 75;606- 616, 1977 
    8. Bonnefoy A, Legrand C: Proteolysis of subendothelial adhesive glycoproteins (fibronectin, thrombospondin, and von Willebrand factor) by plasmin, leukocyte cathepsin G, and elastase. Thromb Res 98;323-332, 2000 
    9. Gupta SK, Pillarisetti K, Aiyar N: CXCR4 undergoes complex lineage and inducing agent-dependent dissociation of expression and functional responsiveness to SDF-1alpha during myeloid differentiation. J Leukoc Biol 70;431-438, 2001 
    10. Collins SJ, Ruscetti FW, Gallagher RE, Gallo RC: Normal functional characteristics of cultured human promyelocytic leukemia cells (HL-60) after induction of differentiation by dimethylsulfoxide. J Exp Med 149;969-974, 1979 
    11. Hauert AB, Martinelli S, Marone C, Niggli V: Differentiated HL-60 cells are a valid model system for the analysis of human neutrophil migration and chemotaxis. Int J Biochem Cell Biol 34;838-854, 2002 
    12. Akiyama SK, Olden K, Yamada KM: Fibronectin and integrins in invasion and metastasis. Cancer Metastasis Rev 14; 173-189, 1995 
    13. Friedl P, Noble PB, Zanker KS: Lymphocyte locomotion in three-dimensional collagen gels. Comparison of three quantitative methods for analysing cell trajectories. J Immunol Methods 165;157-165, 1993 
    14. Friedl P, Noble PB, Zanker KS: T lymphocyte locomotion in a three-dimensional collagen matrix. Expression and function of cell adhesion molecules. J Immunol 154;4973-4985, 1995 
    15. Lewis WH: On the locomotion of the polmorphonuclear neutrophils of the rat in autoplasma cultures. Bull J Hopkins Hosp 55;273, 1934 
    16. Kanegasaki S, Nomura Y, Nitta N, Akiyama S, Tamatani T, Goshoh Y, Yoshida T, Sato T, Kikuchi Y: A novel optical assay system for the quantitative measurement of chemotaxis. J Immunol Methods 282;1-11, 2003 
    17. Murphy PM, Baggiolini M, Charo IF, Hebert CA, Horuk R, Matsushima K, Miller LH, Oppenheim JJ, Power CA: International union of pharmacology. XXII. Nomenclature for chemokine receptors. Pharmacol Rev 52;145-176, 2000 
    18. Tanaka T, Bai Z, Srinoulprasert Y, Yang BG, Hayasaka H, Miyasaka M: Chemokines in tumor progression and metastasis. Cancer Sci 96;317-322, 2005 
    19. Wang AZ, Chen JM, Fisher GW, Wang JC, Diamond HS: Improved in vitro models for assay of rheumatoid synoviocyte chemotaxis. Clin Exp Rheumatol 12;293-299, 1994 
    20. Kim HS KK, Kim GH, Seoh JY: Functional differentiation of HL-60 cells by dimethylsulfoxide and phorbol 12-myristate 13-acetate. J Korean Pediat Soc 42;355- 363, 1999 
    21. Traves SL, Smith SJ, Barnes PJ, Donnelly LE: Specific CXC but not CC chemokines cause elevated monocyte migration in COPD: a role for CXCR2. J Leukoc Biol 76;441-450, 2004 
    22. Entschladen F, Drell TLt, Lang K, Masur K, Palm D, Bastian P, Niggemann B, Zaenker KS: Analysis methods of human cell migration. Exp Cell Res 307;418-426, 2005 
    23. Lariviere B, Rouleau M, Picard S, Beaulieu AD: Human plasma fibronectin potentiates the mitogenic activity of platelet- derived growth factor and complements its wound healing effects. Wound Repair Regen 11;79-89, 2003 
    24. Miyasaka M, Tanaka T: Lymphocyte trafficking across high endothelial venules: dogmas and enigmas. Nat Rev Immunol 4;360-370, 2004 
    25. Breitman TR, Hemmi H, Imaizumi M: Induction by physiological agents of differentiation of the human leukemia cell line HL-60 to cells with functional characteristics. Prog Clin Biol Res 226;215-233, 1986 
    26. Wood W, Faria C, Jacinto A: Distinct mechanisms regulate hemocyte chemotaxis during development and wound healing in Drosophila melanogaster. J Cell Biol 173;405-416, 2006 
    27. Gao Z, Metz WA: Unraveling the chemistry of chemokine receptor ligands. Chem Rev 103;3733-3752, 2003 
    28. Airoldi I, Raffaghello L, Piovan E, Cocco C, Carlini B, Amadori A, Corrias MV, Pistoia V: CXCL12 does not attract CXCR4+ human metastatic neuroblastoma cells: clinical implications. Clin Cancer Res 12;77-82, 2006 
    29. Snyderman R, Altman LC, Hausman MS, Mergenhagen SE: Human mononuclear leukocyte chemotaxis: a quantitative assay for humoral and cellular chemotactic factors. J Immunol 108;857-860, 1972 
    30. Sawada J, Shimizu S, Tamatani T, Kanegasaki S, Saito H, Tanaka A, Kambe N, Nakahata T, Matsuda H: Stem cell factor has a suppressive activity to IgE-mediated chemotaxis of mast cells. J Immunol 174;3626-3632, 2005 
    31. Lewis WH: Locomotion of lymphocytes. Bull J Hopkins Hosp 49;29, 1931 
    32. Shibayama H, Tagawa S, Hattori H, Inoue R, Katagiri S, Kitani T: Laminin and fibronectin promote the chemotaxis of human malignant plasma cell lines. Blood 86;719-725, 1995 

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