A newly established in vitro culture using transgenic Drosophila reveals functional coupling between the phospholipase A2-generated fatty acid cascade and lipopolysaccharide-dependent activation of the immune deficiency (imd) pathway in insect immunity.
Innate immunity is the first line of defence against infectious micro-organisms, and the basic mechanisms of pathogen recognition and response activation are evolutionarily conserved. In mammals, the innate immune response in combination with antigen-specific recognition is required for the activation of adaptive immunity. Therefore, innate immunity is a pharmaceutical target for the development of immune regulators. Here, for the purpose of pharmaceutical screening, we established an in vitro culture based on the innate immune response of Drosophila. The in vitro system is capable of measuring lipopolysaccharide (LPS)-dependent activation of the immune deficiency (imd) pathway, which is similar to the tumour necrosis factor signalling pathway in mammals. Screening revealed that well-known inhibitors of phospholipase A(2) (PLA(2)), dexamethasone (Dex) and p-bromophenacyl bromide (BPB) inhibit LPS-dependent activation of the imd pathway. The inhibitory effects of Dex and BPB were suppressed by the addition of an excess of three (arachidonic acid, eicosapentaenoic acid and gamma-linolenic acid) of the fatty acids so far tested. Arachidonic acid, however, did not activate the imd pathway when used as the sole agonist. These findings indicate that PLA(2) participates in LPS-dependent activation of the imd pathway via the generation of arachidonic acid and other mediators, but requires additional signalling from LPS stimulation. Moreover, PLA(2) was activated in response to bacterial infection in Sarcophaga. These results suggest a functional link between the PLA(2)-generated fatty acid cascade and the LPS-stimulated imd pathway in insect immunity.
Yajima
Masashi
M
Graduate School of Pharmaceutical Sciences, Tohoku University, Aramaki, Aoba-ku, Sendai 980-8578, Japan.
Takada
Masatoshi
M
Takahashi
Nahoko
N
Kikuchi
Haruhisa
H
Natori
Shunji
S
Oshima
Yoshiteru
Y
Kurata
Shoichiro
S
D000098:Acetophenones / Q000494:pharmacology; D000818:Animals; D030801:Animals, Genetically Modified; D016718:Arachidonic Acid / Q000494:pharmacology; D001681:Biological Assay / Q000379:methods*; D003907:Dexamethasone / Q000494:pharmacology; D004175:Diptera / Q000378:metabolism; D004330:Drosophila / Q000235:genetics* / Q000276:immunology*; D029721:Drosophila Proteins / Q000378:metabolism; D004791:Enzyme Inhibitors / Q000494:pharmacology; D005227:Fatty Acids / Q000378:metabolism* / Q000494:pharmacology; D017930:Genes, Reporter; D049452:Green Fluorescent Proteins; D007113:Immunity, Innate / Q000187:drug effects / Q000502:physiology; D019476:Insect Proteins / Q000235:genetics; D007814:Larva; D008070:Lipopolysaccharides / Q000276:immunology* / Q000494:pharmacology; D008164:Luminescent Proteins / Q000235:genetics / Q000378:metabolism; D010741:Phospholipases A / Q000037:antagonists & inhibitors / Q000378:metabolism*; D054467:Phospholipases A2; D015398:Signal Transduction / Q000276:immunology
D000098:Acetophenones; C054756:DptA protein, Drosophila; D029721:Drosophila Proteins; D004791:Enzyme Inhibitors; D005227:Fatty Acids; D019476:Insect Proteins; D008070:Lipopolysaccharides; D008164:Luminescent Proteins; D049452:Green Fluorescent Proteins; D016718:Arachidonic Acid; D003907:Dexamethasone; D010741:Phospholipases A; D054467:Phospholipases A2; C006821:4-bromophenacyl bromide