RESUMO
The purpose of this investigation was to use molecular and immunological techniques to determine whether oral Treponema infected the human brain. Pieces of frontal lobe cortex from 34 subjects were analyzed with species-specific PCR and monoclonal antibodies. PCR detected Treponema in 14/16 Alzheimer's disease (AD) and 4/18 non-AD donors (P < 0.001), and AD specimens had more Treponema species than controls (P < 0.001). PCR also detected Treponema in trigeminal ganglia from three AD and two control donors. Cortex from 15/16 AD subjects and 6/18 controls contained Treponema pectinovorum and/or Treponema socranskii species-specific antigens (P < 0.01). T. pectinovorum and/or T. socranskii antigens were also found in trigeminal ganglia and pons from four embalmed cadavers, and 2/4 cadavers also had Treponema in the hippocampus. These findings suggest that oral Treponema may infect the brain via branches of the trigeminal nerve.
Assuntos
Doença de Alzheimer/microbiologia , Encéfalo/microbiologia , Boca/microbiologia , Treponema/classificação , Adulto , Idoso , Idoso de 80 Anos ou mais , Anticorpos Monoclonais , Borrelia burgdorferi/isolamento & purificação , Cadáver , Distribuição de Qui-Quadrado , Corantes , DNA Bacteriano/genética , Feminino , Lobo Frontal/microbiologia , Hipocampo/microbiologia , Humanos , Técnicas Imunoenzimáticas , Masculino , Pessoa de Meia-Idade , Reação em Cadeia da Polimerase , Ponte/microbiologia , Saliva/microbiologia , Estatísticas não Paramétricas , Treponema/genética , Treponema/imunologia , Gânglio Trigeminal/microbiologia , Nervo Trigêmeo/microbiologiaRESUMO
Identification of plaque spirochetes from dogs is rare and no studies to date report cultivation of canine or feline plaque spirochetes. Plaque samples obtained from canine and feline patients were cultured in broth media. Spirochetes cultured were subjected to microscopic evaluation and were cloned on a solid medium. The clones were provisionally identified using species-specific PCR for treponema isolated from human plaque. Canine spirochete clones included Treponema denticola, T. socranskii ssp., T. vincentii, T. maltophilum, T. medium, and T. pectinovorum. Feline clones included T. maltophilum and T. socranskii. Non-amplified clones may represent novel treponemes. Future studies will be aimed at comparison of the spirochetes present in dogs and cats with or without periodontal disease.
Assuntos
Doenças do Gato/microbiologia , Doenças do Cão/microbiologia , Periodontite/veterinária , Treponema/isolamento & purificação , Infecções por Treponema/veterinária , Animais , Gatos , Cães , Humanos , Periodontite/microbiologia , Reação em Cadeia da Polimerase/veterinária , Treponema/classificação , Treponema/genética , Treponema/ultraestrutura , Infecções por Treponema/microbiologiaRESUMO
Recent findings challenge the assumption that pathogen-related oral spirochetes (PROS) are related to Treponema pallidum. Treponema vincentii, grown in OMIZ-Pat media, cross-reacted with monoclonal antibody H9-2 against T. pallidum, and cultivable PROS had 16S rRNA gene sequences similar to those of T. vincentii (C.-B. Choi, C. Wyss, and U. B. Göbel. J. Clin. Microbiol. 34:1922-1925, 1996). Aims of the present study were to determine whether antigen phenotypes of oral treponemas were influenced by growth conditions and to evaluate the genetic relatedness of cultivable PROS to T. pallidum and T. vincentii. Results show that three T. pallidum monoclonal antibodies (H9-1, H9-2, and F5) cross-reacted with whole cells from four Treponema species grown in modified OMIZ-Pat medium, but not with treponemas grown in NOS medium. Only H9-2 reacted in immunoblots with reduced proteins from cultivable PROS and T. vincentii. Three of five PROS isolates were amplified by T. vincentii-specific PCR, and one was amplified by Treponema medium-specific PCR. None were amplified by T. pallidum-specific PCR. Three of five PROS isolates had 16S ribosomal DNA restriction fragment length polymorphism patterns identical to that of T. vincentii, and the patterns of two isolates resembled that of T. medium. Arbitrarily primed-PCR profiles from whole genomic DNA were distinct among five PROS isolates and two T. vincentii strains. Thus, PROS isolates represent a heterogeneous group of treponemas that share some 16S rRNA gene sequences with T. vincentii and T. medium, but not with T. pallidum. It is proposed that the PROS nomenclature be dropped.
Assuntos
Boca/microbiologia , Spirochaetaceae/classificação , Spirochaetaceae/genética , Treponema/classificação , Treponema/genética , Anticorpos Monoclonais , Antígenos de Bactérias , Técnicas de Tipagem Bacteriana , Sequência de Bases , Reações Cruzadas , Meios de Cultura , Primers do DNA/genética , Genótipo , Humanos , Immunoblotting , Fenótipo , Reação em Cadeia da Polimerase , Polimorfismo de Fragmento de Restrição , RNA Bacteriano/genética , RNA Ribossômico 16S/genética , Especificidade da Espécie , Spirochaetaceae/patogenicidade , Treponema/patogenicidade , Treponema pallidum/classificação , Treponema pallidum/genética , Treponema pallidum/patogenicidadeRESUMO
Species-specific nested PCR was used to detect Treponema amylovorum, Treponema denticola, Treponema maltophilum, Treponema medium, Treponema pectinovorum, Treponema socranskii, and Treponema vincentii in dental plaque. Subjects with periodontitis harbored all species, but T. pectinovorum and T. vincentii were not found in plaque from disease-free subjects.
