Type

Journal Article

Authors

Kevin Kavanagh
Frans B. Wientjes
Julie Renwick
Emer P. Reeves
David Bergin

Subjects

Biochemistry

Topics
nadph oxidase proteins protein binding human matrix assisted laser desorption ionization immune response biology neutrophils insect proteins wax moth galleria mellonella

Superoxide Production in Galleria mellonella Hemocytes: Identification of Proteins Homologous to the NADPH Oxidase Complex of Human Neutrophils (2005)

Abstract The insect immune response has a number of structural and functional similarities to the innate immune response of mammals. The objective of the work presented here was to establish the mechanism by which insect hemocytes produce superoxide and to ascertain whether the proteins involved in superoxide production are similar to those involved in the NADPH oxidase-induced superoxide production in human neutrophils. Hemocytes of the greater wax moth (Galleria mellonella) were shown to be capable of phagocytosing bacterial and fungal cells. The kinetics of phagocytosis and microbial killing were similar in the insect hemocytes and human neutrophils. Superoxide production and microbial killing by both cell types were inhibited in the presence of the NADPH oxidase inhibitor diphenyleneiodonium chloride. Immunoblotting of G. mellonella hemocytes with antibodies raised against human neutrophil phox proteins revealed the presence of proteins homologous to gp91phox, p67phox, p47phox, and the GTP-binding protein rac 2. A protein equivalent to p40phox was not detected in insect hemocytes. Immunofluorescence analysis localized insect 47-kDa and 67-kDa proteins throughout the cytosol and in the perinuclear region. Hemocyte 67-kDa and 47-kDa proteins were immunoprecipitated and analyzed by matrix-assisted laser desorption ionization-time of flight analysis. The results revealed that the hemocyte 67-kDa and 47-kDa proteins contained peptides matching those of p67phox and p47phox of human neutrophils. The results presented here indicate that insect hemocytes phagocytose and kill bacterial and fungal cells by a mechanism similar to the mechanism used by human neutrophils via the production of superoxide. We identified proteins homologous to a number of proteins essential for superoxide production in human neutrophils and demonstrated that significant regions of the 67-kDa and 47-kDa insect proteins are identical to regions of the p67phox and p47phox proteins of neutrophils.
Collections Ireland -> Maynooth University -> Subject = Science & Engineering: Biology
Ireland -> Maynooth University -> Subject = Science & Engineering
Ireland -> Maynooth University -> Academic Unit = Faculty of Science and Engineering: Biology
Ireland -> Maynooth University -> Status = Published
Ireland -> Maynooth University -> Open Access DRIVERset
Ireland -> National University of Ireland Maynooth -> Status = Published
Ireland -> National University of Ireland Maynooth -> Open Access DRIVERset
Ireland -> National University of Ireland Maynooth -> Academic Unit = Faculty of Science and Engineering
Ireland -> National University of Ireland Maynooth -> Academic Unit = Faculty of Science and Engineering: Biology
Ireland -> Maynooth University -> Type = Article
Ireland -> National University of Ireland Maynooth -> Type = Article
Ireland -> Maynooth University -> Academic Unit = Faculty of Science and Engineering

Full list of authors on original publication

Kevin Kavanagh, Frans B. Wientjes, Julie Renwick, Emer P. Reeves, David Bergin

Experts in our system

1
Kevin Kavanagh
Maynooth University
Total Publications: 183
 
2
Emer P Reeves
Royal College of Surgeons in Ireland
Total Publications: 63
 
3
David A Bergin
Royal College of Surgeons in Ireland
Total Publications: 28