Adherence of Human Oral Spirochetes by Collagen‐Binding Proteins

Umemoto, Toshihiko; Li, Mingyu; Namikawa, Isamu

doi:10.1111/j.1348-0421.1997.tb01950.x

Cited by 24 publications

(25 citation statements)

References 31 publications

Supporting

Mentioning

Contrasting

Order By: Relevance

“…These include the virulence factors CNE of Streptococcus equi (29) and Acm of Enterococcus faecium (35), the surface proteins RspA and RspB of Erysipelothrix rhusiopathiae (46), the S-layer protein CbsA of the nonpathogenic bacterium Lactobacillus crispatus (32,49), and the adhesin YadA found in Yersinia enterocolitica and Yersinia pseudotuberculosis (53). Treponema also has the ability to bind collagen directly by collagen-binding polypeptides (54,55).In contrast to the above, previous studies using isolated collagen have been unable to detect a direct interaction between this ECM component and whole B. burgdorferi cells (4,(20)(21)(22). This is despite the fact that B. burgdorferi is mainly an extracellular pathogen that migrates actively from the dermis into the bloodstream and from the blood into connective tissues of organs, such as the heart.…”

contrasting

confidence: 38%

Borrelia burgdorferi Binds to, Invades, and Colonizes Native Type I Collagen Lattices

Zambrano¹,

Beklemisheva²,

Bryksin³

et al. 2004

Infect Immun

View full text Add to dashboard Cite

Borrelia burgdorferi binds strongly to the extracellular matrix and cells of the connective tissue, a binding apparently mediated by specific proteins and proteoglycans. We investigated the interactions between B. burgdorferi cells and intact type I collagen using hydrated lattices that reproduce features of in vivo collagen matrices. B. burgdorferi cells of several strains adhered avidly to these acellular matrices by a mechanism that was not mediated by decorin or other proteoglycans. Moreover, following adhesion to these matrices, B. burgdorferi grew and formed microcolonies. The collagen used in these studies was confirmed to lack decorin by immunoblot analysis; B. burgdorferi cells lacking the decorin adhesin bound readily to intact collagen matrices. B. burgdorferi also bound to collagen lattices that incorporated enzymes that degraded glycosaminoglycan chains in any residual proteoglycans. Binding of the bacteria to intact collagen was nonetheless specific, as bacteria did not bind agar and showed only minimal binding to bovine serum albumin, gelatin, pepsinized type I collagen, and intact collagen that had been misassembled under nonphysiological pH and ionic-strength conditions. Proteinase K treatment of B. burgdorferi cells decreased the binding, as did a lack of flagella, suggesting that surface-exposed proteins and motility may be involved in the ability of B. burgdorferi to interact with intact collagen matrices. The high efficiency of binding of B. burgdorferi strains to intact collagen matrices permits replacement of the commonly used isotopic binding assay with visual fluorescent microscopic assays and will facilitate future studies of these interactions.Borrelia burgdorferi, the etiologic agent of Lyme disease, is transmitted to animals and humans mainly by nymphae of the tick Ixodes scapularis, which during a blood meal deposit a small number of microorganisms into the skin (8,43,51). The inoculation of the bacteria by the tick bite results after a few days in a characteristic rash, erythema migrans, which may be accompanied by systemic symptoms, including malaise, fatigue, fever, headache, neck stiffness, arthralgias, or myalgias (30,51,58). Adhesion, colonization, and proliferation within the skin and other host organs and tissues by B. burgdorferi necessitates interaction between the spirochete and cells of the connective tissue, including macrophages, dendritic cells, fibroblasts (24,48), and the associated extracellular matrix (ECM) (24).B. burgdorferi expresses cell surface proteins that interact specifically with different components of the ECM of the host organism and of mammalian cells in culture (8,23,26,30). These B. burgdorferi surface proteins include the fibronectin receptor encoded by BBK32 (42); proteins that bind directly to glycosaminoglycans (GAGs), such as Bgp (39-41); and membrane lipoproteins, such as DbpA and DbpB, which bind to decorin (4,16,(20)(21)(22). The B. burgdorferi outer membrane protein p66 binds to the beta3 integrin chain of host ECM receptors (9-12). The var...

show abstract

contrasting

confidence: 38%

Borrelia burgdorferi Binds to, Invades, and Colonizes Native Type I Collagen Lattices

Zambrano¹,

Beklemisheva²,

Bryksin³

et al. 2004

Infect Immun

View full text Add to dashboard Cite

show abstract

“…The washed membranes were incubated with 1% skim milk-TBS as blocking solution for 1 hr. The rabbit albumin-binding polypeptides from the treponemal cells were examined by incubation of the treponemal polypeptide-loaded nitrocellulose membranes with PBS, or rabbit albumin solution (120-360 ug/ml), for 2 hr at 4 C, followed by washing with Tween 20-TBS, blocking with skim milk-TBS, and incubation with the horseradish peroxidase (HRP)-conjugated sheep anti-rabbit albumin serum (I: 1,000 dilution; Bethyl Laboratories Inc., Montgomery, Tex., U.S.A.) for 2 hr at room temperature, as described by Umemoto et al (25).…”

Section: Methodsmentioning

confidence: 99%

“…Location of the rabbit albumin-binding polypeptides on the treponemal cells was carried out by immunoelectron microscopy as described by Umemoto et al (25). Briefly, the T. denticola and T. vincentii cells of each 4-day culture were gently washed twice with 0.01 M phosphate-buffered saline (PBS, pH 7.2), fixed for 1 hr at room temperature in 4% paraformaldehyde-0.5% glutaraldehyde fixate (pH 7.0), and washed twice with 1% glycine-PBS.…”

Section: Methodsmentioning

confidence: 99%

Chemotaxis of Oral Treponemes toward Sera and Albumin of Rabbit

Umemoto

Jinno

Taiji

et al. 2001

Microbiology and Immunology

Self Cite

View full text Add to dashboard Cite

Abstract:The motility and chemotaxis of human oral spirochetes Treponemadenticola ATCC 35404, T. medium ATCC 700293, and T. vincentii ATCC 35580 were examined by a capillary assay method. Of five sera three human oral treponemes were dominantly chemoattractant to the rabbit serum. The checkerboard analysis of chemotaxis toward rabbit serum clearly showed that the motile T. denticola cells swam toward the culture media containing higher concentrations of the rabbit serum. T. denticola chemotaxis to the rabbit serum was clearly reduced by heating serum, and rabbit albumin contributed by 60 to 70% to its chemotaxis to the rabbit serum. Western blotting analysis demonstrated that these treponemes possessed rabbit albumin-binding polypeptides with approximate molecular sizes of 65 kDa and 70 kDa. Immunoelectron microscopy demonstrated that a 65 kDa rabbit albumin-binding polypeptide was located on the outer envelopes, suggesting that the rabbit albumin-binding polypeptide is responsible for chemotaxis toward rabbit serum.Key words: Chemotaxis, Oral spirochetes, Serum albumin, Albumin-binding polypeptides Many flagellated microorganisms move either towards or away from chemical stimulants (1,5,27) and possess specific receptors for reacting to the attractants or repellents (2). It has been suggested that flagella may playa role in spirochetal motility (4, 9, 15), because flagelladeficient mutants have been seen to lose their motility (6). In the human oral cavity, anaerobic treponemes are frequently detected in subgingival plaque samples from both healthy and diseased sites (12,23,29). Specific nutrients in sera, such as albumin or fatty acids, seem to be essential requisites for in vitro cultivation of their individual species (28). Chemotaxis toward serum in the crevice fluid would benefit these nutritionally strict treponemes in those limited ecological environments where many kinds of oral microorganisms compete with neighboring organisms for growth nutrients. In this study, we examined the chemotactic characterizations of human oral spirochetes for sera and predominant serum albumin. Materials and MethodsBacterial strains, culture, and growth conditions. Human oral treponemes, T. denticola ATCC 35404, T. medium ATCC 700293, and T. vincentii ATCC 35580, *Addresscorrespondence to Dr. Tomohiko Ogawa, Department of Oral Microbiology, Asahi University School of Dentistry, Hozumi, Hozumi-cho, Motosu-gun, Gifu 501-0296, Japan. Fax: +81-58-329-1422. E-mail: tomo527@dent.asahi-u.ac.jp 571 were used in this study. These spirochetes were all anaerobically grown in trypticase-yeast extract-gelatinevolatile fatty acid-rabbit serum (TYGVS) medium, in an anaerobic chamber (Nz:Hz:COz, 80%: 10%: 10%) at 37 C (16). The treponemes of each 4-day culture were gently washed twice with 10% rabbit serum-free TYGVS medium (TYGV), suspended gently again in the TYGV medium, and counted by a Petroff-Hausser bacteria counter (C.A. Hausser & Son, Pa., U.S.A.). Bacterial cells were suspended in the TYGV (negative control), serum-, or albumin-c...

show abstract

“…Fenno et al (17) also found that recombinant Msp did not show an oligomeric form but it adhered to fibronectin and laminin. Umemoto et al (84,86) reported that T vincentii and T socranskii subsp. huecale have outer envelope proteins that bind to laminin, and that the 95 kDa outer sheath protein of T medium binds to type IV collagen.…”

Section: Adherencementioning

confidence: 99%