title: Highly homologous Mycobacterium tuberculosis chaperonin 60 proteins with differential CD14 dependencies stimulate cytokine production by human monocytes through cooperative activation of p38 and ERK1/2 mitogen-activated protein kinases.
abstract: Tuberculosis is a chronic inflammatory and destructive disease caused by infection with Mycobacterium tuberculosis. We have previously shown that the mycobacterial chaperonin (Cpn)60.1 and 60.2 proteins stimulate human monocytes to secrete pro-inflammatory cytokines. Identification of the cellular mechanisms that contribute to the chronic inflammation characterised by myobacterial infection is therefore of potential therapeutic benefit. In the present study we have investigated the role of the extracellular signal-regulated (ERK1/2) and p38 mitogen-activated protein kinase (MAPK) families in Cpn60-induced cytokine synthesis, and have compared the effects of the bacterial proteins with those of lipopolysaccharide (LPS). Exposure to Cpn60.1, Cpn60.2 or LPS enhanced ERK1/2 activation with increases in phosphorylation evident between 10 and 30 min and maximal after 60-90 min stimulation. Phosphorylation of ERK1/2 in Cpn60-stimulated monocytes was maintained whereas ERK1/2 was rapidly dephosphorylated in LPS-stimulated cells. Exposure to the chaperonins also caused rapid activation of p38(mapk) with kinetics of phosphorylation comparable to those observed in response to LPS. Selective inhibitors of p38(mapk) (SB203580) or of MEK1/2, the direct upstream activator of ERK1/2 (PD98059), reduced the synthesis of IL-1beta, TNFalpha, IL-6 and IL-8 induced by either the chaperonins or LPS. Experiments in which cells were exposed to a combination of both inhibitors led to a nearly complete abrogation of agonist-induced cytokine synthesis. These results show that the p38(mapk) and ERK1/2 signalling pathways are important regulators of the cellular response to mycobacterial chaperonins and that these pathways cooperate to regulate pro-inflammatory cytokine production by human monocytes.
authors.affiliation: Division of Microbial Diseases, UCL Eastman Dental Institute, University College London, UK.
authors.forename: Jo C
authors.initials: JC
authors.lastname: Lewthwaite
authors.forename: Claire E
authors.initials: CE
authors.lastname: Clarkin
authors.forename: Anthony R M
authors.initials: AR
authors.lastname: Coates
authors.forename: Stephen
authors.initials: S
authors.lastname: Poole
authors.forename: Rachel A
authors.initials: RA
authors.lastname: Lawrence
authors.forename: Caroline P D
authors.initials: CP
authors.lastname: Wheeler-Jones
authors.forename: Andrew A
authors.initials: AA
authors.lastname: Pitsillides
authors.forename: Mahavir
authors.initials: M
authors.lastname: Singh
authors.forename: Brian
authors.initials: B
authors.lastname: Henderson
mesh_terms: D002478:Cells, Cultured; D018834:Chaperonin 60 / Q000494:pharmacology*; D016207:Cytokines / Q000096:biosynthesis*; D048049:Extracellular Signal-Regulated MAP Kinases / Q000378:metabolism*; D005419:Flavonoids / Q000494:pharmacology; D006801:Humans; D007093:Imidazoles / Q000494:pharmacology; D007249:Inflammation / Q000378:metabolism; D018950:Lipopolysaccharide Receptors; D009000:Monocytes / Q000187:drug effects* / Q000378:metabolism; D009169:Mycobacterium tuberculosis; D011725:Pyridines / Q000494:pharmacology; D048051:p38 Mitogen-Activated Protein Kinases / Q000037:antagonists & inhibitors / Q000378:metabolism*
chemical_list: D018834:Chaperonin 60; D016207:Cytokines; D005419:Flavonoids; D007093:Imidazoles; D018950:Lipopolysaccharide Receptors; D011725:Pyridines; D048049:Extracellular Signal-Regulated MAP Kinases; D048051:p38 Mitogen-Activated Protein Kinases; C093642:SB 203580; C093973:2-(2-amino-3-methoxyphenyl)-4H-1-benzopyran-4-one