Abstract:
BACKGROUND:Respiratory burst function resulting in the release of reactive oxygen species such as superoxide anion (O2-) from neutrophils is one of the key mechanisms of the innate immune system, and maladaptive control of this mechanism is thought to play a pivotal role in the development of pathologies such as acute lung injury and sepsis. Ovine models of these pathologies are limited by the poor understanding of ovine neutrophil respiratory burst function. RESULTS:Aspects of ovine neutrophil respiratory burst function to be characterised were: i) the maximum rate of O2- generated (Vmax); ii) the time taken to reach Vmax; iii) the total amount of O2- generated during the reaction; and iv) the duration of the reaction. As well as for unstimulated neutrophils, these aspects were also characterised after incubation with a priming agonist (platelet activating factor [PAF], tumour necrosis factor alpha [TNF-alpha] and lipopolysaccharides [LPS]) activating agonists (N-formylmethionyl-leucyl-phenylalanine [fMLP] and phorbol 12-myristate 13-acetate [PMA]) or a combination of a priming and an activating agonist. In the absence of priming or activating agonists, ovine neutrophils displayed a low level of respiratory burst function which was not enhanced by either PAF, TNF-alpha, LPS or fMLP, but was significantly enhanced by PMA. The PMA-induced respiratory burst function was further enhanced by pre-incubation with PAF, but not with TNF-alpha or LPS. By varying the length of pre-incubation with PAF it was demonstrated that this effect decreased as the duration of pre-incubation with PAF increased, and that PAF was enhancing PMA's effects rather than PMA enhancing PAF's effects. CONCLUSION:This study successfully adapted a commonly used method of measuring human neutrophil respiratory burst function to characterise different aspects of ovine neutrophil respiratory burst function. This improved understanding of ovine neutrophils will facilitate the validitation of ovine biomedical models of human pathologies in which neutrophils have been implicated.
journal_name
BMC Immunoljournal_title
BMC immunologyauthors
Tung JP,Fraser JF,Wood P,Fung YLdoi
10.1186/1471-2172-10-25subject
Has Abstractpub_date
2009-05-08 00:00:00pages
25issn
1471-2172pii
1471-2172-10-25journal_volume
10pub_type
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