A rapid two-step procedure for the purification of human peripheral blood basophils to near homogeneity

Clinical and Experimental Allergy : Journal of the British Society for Allergy and Clinical Immunology
B F GibbsF H Falcone

Abstract

Basophils are increasingly utilized as indicators of allergic inflammation and as primary allergic effector cells to study signalling pathways. However, until the present, their enrichment has been time consuming, costly and limited to relatively few specialized laboratories. We have therefore devised a reproducible and rapid method for the purification of human basophils from small quantities of peripheral blood within 1.5 h, which does not require the use of specialized equipment such as elutriators. Human basophils were obtained from healthy volunteers undergoing venipuncture. Heparinized or K3-ethylenediaminetetraacetic acid blood samples were first subjected to centrifugation in Hetasep, directly followed by negative selection using immunomagnetic beads. Basophil morphology and purity were assessed by May-Grünwald staining of cytospins. IgE-mediated histamine release was analysed spectrofluorometrically and IL-4 and IL-13 production by quantitative RT-PCR. CD203c and CD63 surface expression was measured using flow cytometry before and after activation with anti-IgE. Using this protocol, basophils were enriched close to homogeneity in most cases with a mean purity of 99.34+/-0.88% (range 97-100%, n=18) and a mean recovery o...Continue Reading

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Citations

Nov 15, 2008·Nature Immunology·Booki Min
Aug 19, 2010·Hybridoma·Ling YangJie Zhu
Oct 6, 2009·The Journal of Allergy and Clinical Immunology·Franco H FalconeDavid I Pritchard
May 17, 2011·Clinical and Experimental Allergy : Journal of the British Society for Allergy and Clinical Immunology·F H FalconeB F Gibbs
Nov 21, 2012·Experimental Dermatology·Bernhard F GibbsFrancesca Levi-Schaffer
Sep 22, 2009·European Journal of Immunology·Vadim V SumbayevBernhard F Gibbs
Jun 26, 2012·The International Journal of Biochemistry & Cell Biology·Bernhard F GibbsVadim V Sumbayev
Jun 16, 2012·The Journal of Allergy and Clinical Immunology·Natalija NovakCezmi Akdis
Dec 20, 2011·The Journal of Allergy and Clinical Immunology·Markus BöhmUlrike Raap
May 9, 2014·The Journal of Allergy and Clinical Immunology·Sa'ar MizrahiFrancesca Levi-Schaffer
May 13, 2014·The Journal of Allergy and Clinical Immunology·Vito SabatoDidier G Ebo
May 20, 2016·Clinical and Experimental Allergy : Journal of the British Society for Allergy and Clinical Immunology·S KleinerU Raap
Dec 22, 2016·Clinical and Experimental Allergy : Journal of the British Society for Allergy and Clinical Immunology·U RaapB F Gibbs
Feb 13, 2018·Clinical and Experimental Allergy : Journal of the British Society for Allergy and Clinical Immunology·P KorošecP J Turner
Nov 27, 2014·Plastic and Reconstructive Surgery. Global Open·Fumio OhnishiKazuo Kishi
Nov 16, 2018·Current Allergy and Asthma Reports·Oliver HemmingsAlexandra F Santos
Apr 16, 2019·Frontiers in Pharmacology·Inna M YasinskaBernhard F Gibbs
Jan 1, 2021·Frontiers in Immunology·Anuya ParanjapeStephen J Galli

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Basophils are myeloid cells with a high affinity IgE receptor and is involved in inflammatory responses during allergy. Discover the latest research on Basophils here.