PMID: 7540045May 29, 1995Paper

Development of oxidase 'priming' in maturing HL60 cells: correlation with protein expression and tyrosine phosphorylation

Biochimica Et Biophysica Acta
D Lloyds, M B Hallett

Abstract

The mechanisms involved in 'priming' of the oxidase response of neutrophils are unknown. Two major problems are encountered in using circulating neutrophils; firstly, prior exposure to circulating 'priming' cytokines cannot be controlled and secondly, non-intentional 'priming' during cell separation can occur. In this study, these problems were avoided by differentiating the promyeloid leukaemic cell line, HL60, towards granulocytes using dibutyrl cyclic AMP, to produce a 'virgin cell' model system. We have demonstrated that the ability of substance P to both prime the oxidase response and induce tyrosine phosphorylation increased during differentiation. The major tyrosine-phosphorylated protein, with molecular weight of 74 kDa, was not recognised by anti-c-raf1 antibodies. Furthermore, c-raf1 expression rapidly declined during HL60 cell granulocytic differentiation. This data shows that although there was no simple relationship between c-raf quantity and priming, the data were consistent with tyrosine phosphorylation of a 74 kDa protein being important for oxidase 'priming'.

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Citations

Jun 1, 1995·Immunology Today·M B Hallett, D Lloyds
Mar 28, 1997·Journal of Immunological Methods·A al-ShamiP H Naccache
Jun 4, 2008·Journal of Leukocyte Biology·Sabrina Bréchard, Eric J Tschirhart
May 2, 2003·The Journal of Biological Chemistry·Loralee HaughnDavid M Hockenbery

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