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Purification of Basophils from Peripheral Human Blood

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Basophils and Mast Cells

Part of the book series: Methods in Molecular Biology ((MIMB,volume 1192))

Abstract

The purification of basophils from peripheral blood has represented a formidable challenge for researchers since they were discovered by Paul Ehrlich in 1879. From the first published attempts in the late 1960s, it took half a century to develop robust protocols able to provide sufficient numbers of pure, functionally unimpaired basophils. The existing protocols for basophil purification exploit those properties of basophils which distinguish them from other cell types such as their localization in blood, density, and the presence or absence of surface markers. Purification techniques have been used in various combinations and variations to achieve a common goal in mind: to obtain a pure population of human basophils in sufficient numbers for downstream studies. The arduous way leading up to the modern protocols is summarized in this historical retrospective. A fast protocol for purification of basophils to near homogeneity is also described.

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References

  1. Falcone FH, Haas H, Gibbs BF (2000) The human basophil: a new appreciation of its role in immune responses. Blood 96:4028–4038

    CAS  PubMed  Google Scholar 

  2. Sampson D, Archer GT (1967) Release of histamine from human basophils. Blood 29:722–736

    CAS  PubMed  Google Scholar 

  3. Parwaresch MR (1968) A new method for the enrichment of leukocytes, particularly of basophils. Blut 17:260–265

    Article  CAS  PubMed  Google Scholar 

  4. Pruzansky JJ, Patterson R (1970) Decrease in basophils after incubation with specific antigens of leukocytes from allergic donors. Int Arch Allergy Appl Immunol 38:522–526

    Article  CAS  PubMed  Google Scholar 

  5. Day RP (1972) Basophil leucocyte separation from human peripheral blood: a technique for their isolation in high purity and high yield. Clin Allergy 2:205–212

    Article  CAS  PubMed  Google Scholar 

  6. Ishizaka T, De Bernardo R, Tomioka H et al (1972) Identification of basophil granulocytes as a site of allergic histamine release. J Immunol 108:1000–1008

    CAS  PubMed  Google Scholar 

  7. MacGlashan DW, Lichtenstein LM (1980) The purification of human basophils. J Immunol 124:2519–2521

    PubMed  Google Scholar 

  8. Toll JB, Wikberg JE, Andersson RG (1981) Purification of human basophils by affinity chromatography on anti-IgE-sepharose 6 MB. Allergy 36:411–417

    Article  CAS  PubMed  Google Scholar 

  9. Pruzansky JJ, Patterson R (1981) Enrichment of human basophils. J Immunol Methods 44:183–190

    Article  CAS  PubMed  Google Scholar 

  10. Landry FJ, Findlay SR (1983) Purification of human basophils by negative selection. J Immunol Methods 63:329–336

    Article  CAS  PubMed  Google Scholar 

  11. Raghuprasad PK (1982) A rapid simple method of basophil purification by density centrifugation on Percoll. J Immunol 129:2128–2133

    CAS  PubMed  Google Scholar 

  12. Kauffman HF, Levering PR, De Vries K (1983) A single centrifugation step method for the simultaneous separation of different leukocytes with special reference to basophilic leukocytes. J Immunol Methods 57:1–7

    Article  CAS  PubMed  Google Scholar 

  13. Weil GJ, Leiserson WM, Chused TM (1983) Isolation of human basophils by flow microfluorometry. J Immunol Methods 58:359–363

    Article  CAS  PubMed  Google Scholar 

  14. Bjerke T, Nielsen S, Helgestad J et al (1993) Purification of human blood basophils by negative selection using immunomagnetic beads. J Immunol Methods 157:49–56

    Article  CAS  PubMed  Google Scholar 

  15. Nielsen HV, Shah PM, Schiøtz PO (1998) Factors determining spontaneous histamine release from human basophils purified with Percoll gradients and Dynabeads. Allergy 53:302–306

    Article  CAS  PubMed  Google Scholar 

  16. Pendy LM, Arra SJ, Anselmino LM, Thomas LL (1986) A flow cytometric procedure for the isolation of antigenically responsive human basophils. J Immunol Methods 91:59–63

    Article  CAS  PubMed  Google Scholar 

  17. Pruzansky JJ, Grammer LC, Patterson R, Roberts M (1983) Dissociation of IgE from receptors on human basophils. I. Enhanced passive sensitization for histamine release. J Immunol 131:1949–1953

    CAS  PubMed  Google Scholar 

  18. Bodger MP, Newton LA (1987) The purification of human basophils: their immunophenotype and cytochemistry. Br J Haematol 67:281–284

    Article  CAS  PubMed  Google Scholar 

  19. Gibbs BF, Noll T, Falcone FH et al (1997) A three-step procedure for the purification of human basophils from buffy coat blood. Inflamm Res 46:137–142

    Article  CAS  PubMed  Google Scholar 

  20. Haisch K, Gibbs BF, Körber H et al (1999) Purification of morphologically and functionally intact human basophils to near homogeneity. J Immunol Methods 226:129–137

    Article  CAS  PubMed  Google Scholar 

  21. De Boer M, Roos D (1986) Metabolic comparison between basophils and other leukocytes from human blood. J Immunol 136:3447–3454

    PubMed  Google Scholar 

  22. Splinter TA, Beudeker M, Van Beek A (1978) Changes in cell density induced by isopaque. Exp Cell Res 111:245–251

    Article  CAS  PubMed  Google Scholar 

  23. Graham HT, Lowry OH, Wheelwright F et al (1955) Distribution of histamine among leukocytes and platelets. Blood 10:467–481

    CAS  PubMed  Google Scholar 

  24. Miroli AA, James BM, Spitz M (1986) Single step enrichment of human peripheral blood basophils by Ficoll-Paque centrifugation. J Immunol Methods 88:91–96

    Article  CAS  PubMed  Google Scholar 

  25. Lett-Brown MA, Robinson L, Juneja HS, Grant JA (1989) Purification of human basophils. Their response to anti-IgE. J Immunol Methods 117:163–167

    Article  CAS  PubMed  Google Scholar 

  26. Schroeder JT, Hanrahan LR (1990) Purification of human basophils using mouse monoclonal IgE. J Immunol Methods 133:269–277

    Article  CAS  PubMed  Google Scholar 

  27. Mul FP, Knol EF, Roos D (1992) An improved method for the purification of basophilic granulocytes from human blood. J Immunol Methods 149:207–214

    Article  CAS  PubMed  Google Scholar 

  28. Mita H, Akiyama K, Hayakawa T et al (1993) Purification of human blood basophils and leukotriene C4 generation following calcium ionophore stimulation. Prostaglandins Leukot Essent Fatty Acids 49:783–788

    Article  CAS  PubMed  Google Scholar 

  29. Falcone FH, Dahinden CA, Gibbs BF et al (1996) Human basophils release interleukin-4 after stimulation with Schistosoma mansoni egg antigen. Eur J Immunol 26:1147–1155

    Article  CAS  PubMed  Google Scholar 

  30. Tanimoto Y, Takahashi K, Takata M et al (1992) Purification of human blood basophils using negative selection by flow cytometry. Clin Exp Allergy 22:1015–1019

    Article  CAS  PubMed  Google Scholar 

  31. Kepley C, Craig S, Schwartz L (1994) Purification of human basophils by density and size alone. J Immunol Methods 175:1–9

    Article  CAS  PubMed  Google Scholar 

  32. Willheim M, Agis H, Sperr WR et al (1995) Purification of human basophils and mast cells by multistep separation technique and mAb to CDw17 and CD117/c-kit. J Immunol Methods 182:115–129

    Article  CAS  PubMed  Google Scholar 

  33. Tsang S, Hayashi M, Zheng X et al (2000) Simplified purification of human basophils. J Immunol Methods 233:13–20

    Article  CAS  PubMed  Google Scholar 

  34. Gibbs BF, Papenfuss K, Falcone FH (2008) A rapid two-step procedure for the purification of human peripheral blood basophils to near homogeneity. Clin Exp Allergy 38:480–485

    Article  CAS  PubMed  Google Scholar 

  35. Gilbert HS, Ornstein L (1975) Basophil counting with a new staining method using alcian blue. Blood 46:279–286

    CAS  PubMed  Google Scholar 

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Acknowledgements

This work was supported by the COST Action BM1007 Mast Cells and Basophils—Targets for Innovative Therapies.

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Authors and Affiliations

  1. Division of Molecular and Cellular Science, The School of Pharmacy, The University of Nottingham, Boots Science Building, Science Road, Nottingham, NG7 2RD, UK

    Franco H. Falcone

  2. Medway School of Pharmacy, The University of Kent, Chatham Maritime, Kent, UK

    Bernhard F. Gibbs

Authors
  1. Franco H. Falcone
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  2. Bernhard F. Gibbs
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Corresponding author

Correspondence to Franco H. Falcone .

Editor information

Editors and Affiliations

  1. Medway School of Pharmacy, The University of Kent, Chatham Maritime, Kent, United Kingdom

    Bernhard F. Gibbs

  2. Division of Molecular and Cellular Science, The School of Pharmacy, The University of Nottingham, Nottingham, United Kingdom

    Franco H. Falcone

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Falcone, F.H., Gibbs, B.F. (2014). Purification of Basophils from Peripheral Human Blood. In: Gibbs, B., Falcone, F. (eds) Basophils and Mast Cells. Methods in Molecular Biology, vol 1192. Humana Press, New York, NY. https://doi.org/10.1007/978-1-4939-1173-8_2

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  • DOI: https://doi.org/10.1007/978-1-4939-1173-8_2

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  • Publisher Name: Humana Press, New York, NY

  • Print ISBN: 978-1-4939-1172-1

  • Online ISBN: 978-1-4939-1173-8

  • eBook Packages: Springer Protocols

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