Detection of heat shock proteins but not superantigen by isolated oral bacteria from patients with Behcet's disease

Abstract

We isolated bacteria from periodontal sites and mixed saliva in eight patients with Behcet's disease and surveyed them to determine whether they produced heat shock proteins (HSPs) and superantigen. Cultivable bacterial compositions from periodontal sites and saliva were examined by anaerobic culture using blood agar plates. Gram-negative anaerobic rods such as Prevotella intermedia, Fusobacterium nucleatum, and Capnocytophaga species were predominant in the isolates from the subgingival plaque samples. The Streptococcus mitis group was the most common type isolated from the saliva samples. To detect the production of HSPs, Western blot analyses were performed using a polyclonal rabbit antibody to Escherichia coli DnaK and a monoclonal antibody to Helicobacter pylori Gro-EL. Sonic extracts of 27 of the strains (79.4%) reacted with the antibody against E. coli DnaK. Nine of these 34 strains (26.5%) were found to produce HSPs that reacted with antibody to H. pylori Gro-EL. A total of 54 isolates were examined for superantigen activity against human peripheral leukocytes. Twenty-five gram-negative clinical strains isolated from chronic periodontitis lesions and 20 ATCC strains of microorganisms were also examined. We could not detect any superantigen activity in 500x diluted supernatant of the strains isolated from the eight patients with Behcet's disease. The present study indicates that the anaerobic strains isolated from the oral cavity of these patients produce HSPs, the production being related to Bechet's disease.

Behcet’s disease: from heat shock proteins to infections

“…Kirino et al hypothesized that HSP60 stimulates not only antigen-specific autoimmune responses, but also the innate immune system through constitutively overexpressed TLR4 [73]. These data are consistent with the interpretation that the immune response against TLR4 ligands, such as HSP and LPS, plays an important part in development of BD [76]. By means of western blot analyses, the bacterial composition of the subgingival plaque and saliva collected from patients with BD was investigated and the levels of superantigen and HSP production by the oral bacteria isolated from these patients (groups of S. salivarius and S. mitis as predominant strains, Prevotella spp, Fusobacterium spp) was determined.…”

“…By means of western blot analyses, the bacterial composition of the subgingival plaque and saliva collected from patients with BD was investigated and the levels of superantigen and HSP production by the oral bacteria isolated from these patients (groups of S. salivarius and S. mitis as predominant strains, Prevotella spp, Fusobacterium spp) was determined. The results indicated that the anaerobic strains isolated from the oral cavity of these patients produce HSPs and possibly that HSPs produced by microorganisms in the oral cavity might lead to the acceleration of oral membrane ulceration in patients with BD [76]. Besides their immunodominance as microbial antigens, under various circumstances HSP do elicit immune responses also when (over-)expressed as self antigens by cells or tissues.…”

“…In some studies, the expression of TLR1, TLR2, TLR4, and TLR6 on monocytes and granulocytes in patients with BD was investigated, which showed increased expression of TLR4, but not TLR2 and others, by peripheral blood mononuclear cells [73,74]. Miura et al [76] hypothesized that TLR4 polymorphisms may be associated with the risk of BD. Kirino et al hypothesized that HSP60 stimulates not only antigen-specific autoimmune responses, but also the innate immune system through constitutively overexpressed TLR4 [73].…”

The role of infections in Behçet disease and neuro-Behçet syndrome

“…Streptococci are more frequently isolated from the oral cavity of people with BD, who also have more frequent streptococcal infections. Initially, Streptococcus sanguis was the identified strain but others have been isolated [28,29]. We hypothesized that if BD is triggered by a streptococcal infection, anti-neuronal antibodies could mediate some neurological involvement and/or constitute a biomarker to help clinical therapeutic decisions.…”

Infection and Behçet Disease

“…The results showed that S. sanguinis infection and the sera from patients with BD are both inflammatory stimuli for heterogeneous nuclear ribonucleoprotein. 20 Once again this process refers to molecular mimicry. 13 However, the stimulation of T cells in patients with BD is not specific for this bacteria and is shared with other species, 14 leading to the hypothesis of a non-specific lymphocytic cell hypersensitivity.…”