Heat shock protein 10 of Chlamydophila pneumoniae induces proinflammatory cytokines through Toll-like receptor (TLR) 2 and TLR4 in human monocytes THP-1.

Inflammatory response is the first line of infection. Previous studies have suggested that Chlamydophila pneumoniae heat shock protein (CHSP) 60 is present in human atheromata, and it plays an important role on the chronic infection elicited by C. pneumoniae. Here, we demonstrated in vitro the impact of heat shock protein 10 (HSP10) of C. pneumoniae on THP-1 cells and the role of Toll-like receptors (TLRs) in the procedures of inflammatory response. The production of proinflammatory cytokines, including tumor necrosis factor alpha (TNF-alpha), interleukin (IL)-6, and IL-1beta were induced by recombinant HSP10 dose-dependently, and the proinflammatory activity of HSP10 was greatly reduced by heating and deproteinization treatment. The expression of TLR4 and TLR2 on the cultured cells were determined by reverse transcriptase-polymerase chain reaction and immunofluorescence. Peritoneal macrophages isolated from wild-type (C3H/HeN) and TLR4-deficient mice (C3H/HeJ) were respectively stimulated with endotoxin-free proteins. Cytokine responses after stimulation were significantly different, depending on the presence of TLR4. The effect on cytokine expression was blocked by anti-TLR2 or anti-TLR4 MAb partially or dramatically. Thus, HSP10 of C. pneumoniae which could elicit inflammatory reactions in human monocytes may contribute to the inflammatory processes in Chlamydophila infection, and the effects were mediated by TLR4 and, to a lesser extent, TLR2.

Results of a phase IIa clinical trial of an anti-inflammatory molecule, chaperonin 10, in multiple sclerosis.

BACKGROUND: Chaperonin 10 (Cpn10) is a mitochondrial molecule involved in protein folding. The aim of this study was to determine the safety profile of Cpn10 in patients with multiple sclerosis (MS). METHODS: A total of 50 patients with relapse-remitting or secondary progressive MS were intravenously administered 5 mg or 10 mg of Cpn10 weekly for 12 weeks in a double-blind, randomized, placebo controlled, phase II trial. Clinical reviews, including Expanded Disability Status Scale and magnetic resonance imaging (MRI) with Gadolinium, were undertaken every 4 weeks. Stimulation of patient peripheral blood mononuclear cells with lipopolysaccharide ex vivo was used to measure the in vivo activity of Cpn10. RESULTS: No significant differences in the frequency of adverse events were seen between treatment and placebo arms. Leukocytes from both groups of Cpn10-treated patients produced significantly lower levels of critical proinflammatory cytokines. A trend toward improvement in new Gadolinium-enhancing lesions on MRI was observed, but this difference was not statistically significant. No differences in clinical outcome measures were seen. CONCLUSIONS: Cpn10 is safe and well tolerated when administered to patients with MS for 3 months, however, a further extended phase II study primarily focused on efficacy is warranted.

Recombinant GroES in combination with CpG oligodeoxynucleotides protects mice against Mycobacterium avium infection.

The groES gene of Mycobacterium avium strain 485 was cloned and expressed in Escherichia coli and the recombinant GroES protein was purified by affinity chromatography. The GroES preparation showed high purity by electrophoresis and immunoblotting. Immuno-electron microscopy showed that GroES was located both in the cytoplasm and on the surface of the mycobacterial cells and thus is readily available to interact with the host immune system. BALB/c mice were immunised intranasally with recombinant GroES, alone or in combination with a synthetic oligodeoxynucleotide containing unmethylated CpG motifs, and tested for protection against infection with M. avium. Neither GroES nor CpG alone provided any protection against subsequent challenge with M. avium, whereas a combination of the two significantly protected the lungs and spleen against colonisation by M. avium after intranasal challenge with a low dose of the organism. This indicates that intranasal administration of GroES and CpG oligodeoxynucleotides increases the resistance of BALB/c mice to M. avium infection.

Preventive administration of Mycobacterium tuberculosis 10-kDa heat shock protein (hsp10) suppresses adjuvant arthritis in Lewis rats.

Adjuvant arthritis (AA) can be induced in Lewis rats by immunization with Mycobacterium tuberculosis (Mt) in oil. We have investigated the modulation of AA by mycobacterial 10-kDa heat shock protein (hsp10), administered according to several protocols known to induce immune tolerance and immune deviation. Subcutaneous immunization with hsp10 in aqueous solution did not induce a cellular immune response, evaluated as delayed-type hypersensitivity (DTH) reaction, although anti-hsp10 antibodies, mainly of the IgG2a isotype, were detected in serum of treated animals. When rats were pretreated with hsp10 in aqueous solution before AA induction, no effects were seen on arthritis-induced joint swelling, although osteolysis and lymphocyte infiltration were slightly decreased. When other routes of administration were attempted, the strongest suppression was seen in the group of animals which received four intranasal (i.n.) administrations of protein and a subsequent challenge of hsp10 in incomplete Freund's adjuvant (IFA). We also found that the extent of disease suppression among the different groups of animals correlated with serum anti-hsp10 antibody levels. These antibodies mostly belonged to the IgG2a subtype, suggesting that immune deviation may play a role in the mechanism of disease suppression by hsp10.