Eosinophils pp 101-110 | Cite as

Eosinophil Chemotaxis

  • Gordon DentEmail author
Part of the Methods in Molecular Biology book series (MIMB, volume 1178)


Chemotaxis assays have a number of applications in the study of leukocyte biology and immune/inflammatory pathology. Multiwell “blind chamber”-type assays allow a large number of parallel measurements within a single assay. The development of fluorescence assays using microplate-based chemotaxis chambers has permitted a degree of automation to be applied to these assays. Here, a method is described for the quantitative measurement of eosinophil migration using a 96-well assay with numbers of cells that may realistically be obtained from blood samples.

Key words

Cell-based assays Chemotaxis Eosinophils Fluorescence Leukocytes Migration 


  1. 1.
    Dent G, Hadjicharalambous C, Yoshikawa T et al (2004) Contribution of eotaxin-1 to eosinophil chemotactic activity of moderate and severe asthmatic sputum. Am J Respir Crit Care Med 169:1110–1117PubMedCrossRefGoogle Scholar
  2. 2.
    Furukawa E, Ohrui T, Yamaya M et al (2004) Human airway submucosal glands augment eosinophil chemotaxis during rhinovirus infection. Clin Exp Allergy 34:704–711PubMedCrossRefGoogle Scholar
  3. 3.
    Fujishima H, Fukagawa K, Okada N et al (2013) Chemotactic responses of peripheral blood eosinophils to prostaglandin D2 in atopic keratoconjunctivitis. Ann Allergy Asthma Immunol 111:126–131PubMedCrossRefGoogle Scholar
  4. 4.
    Hasan AM, Mourtada-Maarabouni M, Hameed MS, Williams GT, Dent G (2010) Phosphoinositide 3-kinase γ mediates chemotactic responses of human eosinophils to platelet-activating factor. Int Immunopharmacol 10:1017–1021PubMedCrossRefGoogle Scholar
  5. 5.
    Heit B, Liu L, Colarusso P, Puri KD, Kubes P (2008) PI3K accelerates, but is not required for, neutrophil chemotaxis to fMLP. J Cell Sci 121:205–214PubMedCrossRefGoogle Scholar
  6. 6.
    Boyden S (1962) The chemotactic effect of mixtures of antibody and antigen on polymorphonuclear leucocytes. J Exp Med 115:453–466PubMedCentralPubMedCrossRefGoogle Scholar
  7. 7.
    Dent G, Hosking LA, Lordan JL et al (2002) Differential roles of IL-16 and CD28/B7 costimulation in the generation of T-lymphocyte chemotactic activity in the bronchial mucosa of mild and moderate asthmatic individuals. J Allergy Clin Immunol 110:906–914PubMedCrossRefGoogle Scholar
  8. 8.
    Hadjicharalambous C, Dent G, May RD et al (2004) Measurement of eotaxin (CCL11) in induced sputum supernatants: validation and detection in asthma. J Allergy Clin Immunol 113:657–662PubMedCrossRefGoogle Scholar
  9. 9.
    Al-Rabia MW, Blaylock MG, Sexton DW, Walsh GM (2004) Membrane receptor-mediated apoptosis and caspase activation in the differentiated Eol-1 eosinophilic cell line. J Leukoc Biol 75:1045–1055Google Scholar
  10. 10.
    Baumann MA, Paul CC (1998) The AML14 and AML14.3D10 cell lines: long overdue model for the study of eosinophils and more. Stem Cells 16:16–24PubMedCrossRefGoogle Scholar
  11. 11.
    Denholm EM, Stankus GP (1995) Differential effects of two fluorescent probes on macrophage migration as assessed by manual and automated methods. Cytometry 19:366–369PubMedCrossRefGoogle Scholar
  12. 12.
    Sunder-Plassmann G, Hofbauer R, Sengoelge G, Hörl WH (1996) Quantification of leukocyte migration: improvement of a method. Immunol Invest 25:49–63PubMedCrossRefGoogle Scholar
  13. 13.
    Taylor ML, Dastych J, Sehgal D et al (2001) The Kit-activating mutation D816V enhances stem cell factor-dependent chemotaxis. Blood 98:1195–1199PubMedCrossRefGoogle Scholar
  14. 14.
    Hasan AM (1999) Intracellular signalling in eosinophils: differential roles for phosphoinositide 3-kinase and p38 MAP kinase in cellular responses to chemoattractants. PhD thesis, Keele UniversityGoogle Scholar
  15. 15.
    Suominen I (2009) Application note AN-HP-REVPIPET-0909: dispense liquids containing proteins more reliably with reverse pipetting. ~ .pdf. Accessed 2 Dec 2013
  16. 16.
    Life Technologies Corp (2013) Calcein, AM, cell-permeant dye. Accessed 2 Dec 2013

Copyright information

© Springer Science+Business Media, New York 2014

Authors and Affiliations

  1. 1.Institute of Science & Technology in MedicineKeele UniversityKeele, StaffordshireUK

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