Application of Intracellular Alkaline Phosphatase Activity Measurement in Detection of Neutrophil Adherence In Vitro

Bednarska, Katarzyna; Klink, Magdalena; Sulowska, Zofia

doi:https://doi.org/10.1155/MI/2006/19307

Mediators of Inflammation

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Research Communication | Open Access

Volume 2006 | Article ID 019307 | https://doi.org/10.1155/MI/2006/19307

Application of Intracellular Alkaline Phosphatase Activity Measurement in Detection of Neutrophil Adherence In Vitro

Katarzyna Bednarska,¹Magdalena Klink,¹and Zofia Sulowska¹

Received28 Apr 2006

Accepted13 Jul 2006

Published06 Sept 2006

Abstract

We have proposed the use of the fluorimetric method with 4-methylumbelliferyl phosphate (4-MUP) specific substrate for the alkaline phosphatase determination in the neutrophil adhesion assay. We provide evidence that the endogenous neutrophil alkaline phosphatase (NAP) activity evaluation is reliable to quantify neutrophil adhesion at a wide range of cell numbers (104–106). The results obtained by fluorimetric NAP activity test correlate to the results of adherence evaluated using the MTT reduction assay. The fluorimetric NAP activity test may be applied for resting as well as activated neutrophils without the risk of the activators interferences into the test. The alkaline phosphatase survey with the use of 4-MUP substrate is recommended herein as a sensitive, repeatable, simple, and reliable method of the neutrophil adherence determination in vitro.

References

M. A. Jutila, “Leukocyte traffic to sites of inflammation,” APMIS, vol. 100, no. 3, pp. 191–201, 1992.
View at: Google Scholar
O. Sørensen and N. Borregaard, “Methods for quantitation of human neutrophil proteins, a survey,” Journal of Immunological Methods, vol. 232, no. 1-2, pp. 179–190, 1999.
View at: Publisher Site | Google Scholar
S. Oez, K. Welte, E. Platzer, and J. R. Kalden, “A simple assay for quantifying the inducible adherence of neutrophils,” Immunobiology, vol. 180, no. 4-5, pp. 308–315, 1990.
View at: Google Scholar
M. Klink, M. Cedzyński, A. St Swierzko, H. Tchórzewski, and Z. Sułowska, “Involvement of nitric oxide donor compounds in the bactericidal activity of human neutrophils in vitro,” Journal of Medical Microbiology, vol. 52, no. 4, pp. 303–308, 2003.
View at: Publisher Site | Google Scholar
T. Mosmann, “Rapid colorimetric assay for cellular growth and survival: application to proliferation and cytotoxicity assays,” Journal of Immunological Methods, vol. 65, no. 1-2, pp. 55–63, 1983.
View at: Publisher Site | Google Scholar
M. Pozzolini, S. Scarfí, U. Benatti, and M. Giovine, “Interference in MTT cell viability assay in activated macrophage cell line,” Analytical Biochemistry, vol. 313, no. 2, pp. 338–341, 2003.
View at: Publisher Site | Google Scholar
C. Rollino, S. Borsa, G. Bellone, G. Piccoli, and G. Emanuelli, “False positive results with MTT assay,” Journal of Immunological Methods, vol. 185, no. 1, pp. 141–143, 1995.
View at: Publisher Site | Google Scholar
A. L. Akeson and C. W. Woods, “A fluorometric assay for the quantitation of cell adherence to endothelial cells,” Journal of Immunological Methods, vol. 163, no. 2, pp. 181–185, 1993.
View at: Publisher Site | Google Scholar
Y. Maeda, K. Tanaka, Y. Koga et al., “A simple quantitative in vitro assay for thymocyte adhesion to thymic epithelial cells using a fluorescein diacetate,” Journal of Immunological Methods, vol. 157, no. 1-2, pp. 117–123, 1993.
View at: Publisher Site | Google Scholar
M. Heimbürger, R. Lerner, and J. Palmblad, “Effects of antirheumatic drugs on adhesiveness of endothelial cells and neutrophils,” Biochemical Pharmacology, vol. 56, no. 12, pp. 1661–1669, 1998.
View at: Publisher Site | Google Scholar
L. S. De Clerck, C. H. Bridts, A. M. Mertens, M. M. Moens, and W. J. Stevens, “Use of fluorescent dyes in the determination of adherence of human leucocytes to endothelial cells and the effect of fluorochromes on cellular function,” Journal of Immunological Methods, vol. 172, no. 1, pp. 115–124, 1994.
View at: Publisher Site | Google Scholar
E. Tolosa and S. Shaw, “A fluorogenic assay of endogenous phosphatase for assessment of cell adhesion,” Journal of Immunological Methods, vol. 192, no. 1-2, pp. 165–172, 1996.
View at: Publisher Site | Google Scholar
L. I. Huschtscha, F. C. Lucibello, and W. F. Bodmer, “A rapid micro method for counting cells “in situ” using a fluorogenic alkaline phosphatase enzyme assay,” In Vitro Cellular & Developmental Biology, vol. 25, no. 1, pp. 105–108, 1989.
View at: Google Scholar
H. Vergnes, J. Grozdea, and J. Corberand, “Liver- and placenta-like alkaline phosphatases (EC 3.1.3.1) isoenzymes in normal human neutrophils,” Enzyme, vol. 31, no. 3, pp. 154–165, 1984.
View at: Google Scholar
N. Borregaard, L. J. Miller, and T. A. Springer, “Chemoattractant-regulated mobilization of a novel intracellular compartment in human neutrophils,” Science, vol. 237, no. 4819, pp. 1204–1206, 1987.
View at: Google Scholar
N. Borregaard, L. Kjeldsen, H. Sengelov et al., “Changes in subcellular localization and surface expression of L-selectin, alkaline phosphatase, and Mac-1 in human neutrophils during stimulation with inflammatory mediators,” Journal of Leukocyte Biology, vol. 56, no. 1, pp. 80–87, 1994.
View at: Google Scholar
G. F. Santini, S. Battistin, P. De Paoli, D Villalta, M Boran, and G Basaglia, “A method for the determination of the adherence of granulocytes to microtitre plates,” Journal of Immunological Methods, vol. 100, no. 1-2, pp. 153–156, 1987.
View at: Publisher Site | Google Scholar
I. M. Roberts, S. L. Jones, R. R. Premier, and J. C. Cox, “A comparison of the sensitivity and specificity of enzyme immunoassays and time-resolved fluoroimmunoassay,” Journal of Immunological Methods, vol. 143, no. 1, pp. 49–56, 1991.
View at: Publisher Site | Google Scholar
K. R. Gee, W.-C. Sun, M. K. Bhalgat et al., “Fluorogenic substrates based on fluorinated umbelliferones for continuous assays of phosphatases and $β$ -galactosidases,” Analytical Biochemistry, vol. 273, no. 1, pp. 41–48, 1999.
View at: Publisher Site | Google Scholar
B. Caughey, L. de Marco, T. J. Peters, K. Mashiter, and W. A. Gibbons, “Analytical subcellular fractionation of rat pituitary homogenates with special reference to the subcellular localization and properties of alkaline phosphatases,” Biochimica et Biophysica Acta, vol. 757, no. 3, pp. 296–301, 1983.
View at: Google Scholar
S. A. Kelly, P. J. Goldschmidt-Clermont, E. E. Milliken, T. Arai, E. H. Smith, and G. B. Bulkley, “Protein tyrosine phosphorylation mediates TNF-induced endothelial-neutrophil adhesion in vitro,” American Journal of Physiology. Heart and Circulatory Physiology, vol. 274, no. 2 pt 2, pp. H513–H519, 1998.
View at: Google Scholar
T. Kobayashi, V. S. Zinchuk, T. Okada, E. G. Del Saz, and H. Seguchi, “Intracellular dynamics of alkaline phosphatase-containing granules in electropermeabilized human neutrophils,” Histochemistry and Cell Biology, vol. 110, no. 4, pp. 395–406, 1998.
View at: Publisher Site | Google Scholar
L. A. Boxer and J. E. Smolen, “Neutrophil granule constituents and their release in health and disease,” Hematology/Oncology Clinics of North America, vol. 2, no. 1, pp. 101–134, 1988.
View at: Google Scholar
V. Fait, S. Sela, E. Ophir et al., “Peripheral polymorphonuclear leukocyte priming contributes to oxidative stress in early pregnancy,” Journal of the Society for Gynecologic Investigation, vol. 12, no. 1, pp. 46–49, 2005.
View at: Publisher Site | Google Scholar
F Stagno, P Guglielmo, U Consoli, P Fiumara, G S Longo, and R Giustolisi, “All-trans-retinoic-acid- and growth-factor- mediated induction of alkaline phosphatase activity in freshly isolated chronic myeloid leukemia cells,” Acta Haematologica, vol. 102, no. 2, pp. 61–65, 1999.
View at: Publisher Site | Google Scholar
A. Karlsson, L. Khalfan, C. Dahlgren, T. Stigbrand, and P. Follin, “Neutrophil alkaline phosphatase activity increase in bacterial infections is not associated with a general increase in secretory vesicle membrane components,” Infection and Immunity, vol. 63, no. 3, pp. 911–916, 1995.
View at: Google Scholar

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Copyright © 2006 Katarzyna Bednarska et al. This is an open access article distributed under the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly cited.

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