Biological activity of a lipopolysaccharide extracted from Treponema hyodysenteriae

Nuessen, M E; Birmingham, J R; Joens, Lynn A.

doi:10.1128/iai.37.1.138-142.1982

Cited by 32 publications

(14 citation statements)

References 18 publications

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“…suggested the possible involvement of endotoxinlike activity associated with those organisms (3,5,10,11,19). However, no precise chemical characterization was performed in these studies.…”

mentioning

confidence: 96%

Absence of lipopolysaccharide in the Lyme disease spirochete, Borrelia burgdorferi

Takayama¹,

Rothenberg²,

Barbour³

1987

Infect Immun

220

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We were unable to deMonstrate the presence of the classic enterobacterium-type lipopolysaccharide in the cells of the Lyme disease spirochete, Borrelia burgdorferi B31. This finding was primarily based on chemical analysis and the absence of free lipid A upon mild acid hydrolysis of the appropriate cell extracts. These re4ults do not preclude the possible existence of an unusual lipopolysaccharide-like compound(s) in B. burgdorferi. Beck et al. (4) recently reported the presence of lipopolysaccharide (LPS) irq the Lyme disease spirochete (Borrelia burgdorferi). We wanted to confirm their results and then examine the nature of the lipid A moiety of LPS from this source. We extracted the cells of B. burgdorferi B31 by two well-known methods used in the isolation of LPS from gram-negative bacteria. The appropriate fractions obtained by these two methods were hydrolyzed under mild acid conditions and examined by thin-layer chromatography (TLC) for the presence of free lipid A. Chemical analyses for the presence of 2-keto-3-deoxyoctonate, glucosamine, and hydroxy fatty acids were performed on the extracts. We found no evidence for the presence of lipid A in any of the fractions tested. Thus, we must conclude that the classic LPS associated with gram-negative bacteria is absent in cells of B. burgdorferi.Cells of B. burgdorferi B31 (ATCC 35210) were grown in BSK II medium (1), washed three times with 50 mM Tris (pH 7.4)-150 mM NaCI-5 mM MgCI2 (2), and lyophilized. In one experiment, the extraction method of Galanos et al. (7) with modifications was used (12,14). The results of this extraction are summarized in Table 1. Almost all of the extractable material was found in the 90% aqueous ethanol fraction, whereas very little material appeared in the phenol-waterchloroform-petroleum ether fraction, which should contain the rough-type LPS (R-LPS). This latter fraction was hydrolyzed in 0.1 N HCI as previously described for the preparation of monophosphoryl lipid A (14), and 30 ,ug of the organic extract was analyzed by TLC for the presence of lipid A (12,14). A mixture of hexa-, penta-, and tetraacyl monopliosphoryl lipid A (from LPS of Salmonella typhimurium) was used as a chromatographic standard (13). There was no evidence of the presence of free lipid A in this sample, indicating the absence of R-LPS in the original extract (phenol-water-chloroform-petroleum ether).Previous studies of lipids X and Y (monosaccharide precursors of LPS) showed that much of X and Y was extracted into the aqueous ethanol fraction (15,16). Thus, it is conceivable that the 113.2 mg of the 90% ethanol extract (Table 1) might contain the R-LPS. This material was initially fractionated by solvent precipitation to yield 79.9 mg of an acetone-insoluble glycolipid-phospholipid fraction. This * Corresponding author. fraction was further purified by preparative TLC on Silica Gel G (Merck & Co., Inc.) by a solvent system of chloroform-methanol-water (65:25:4). Five major fractions (designated A to E) were obtained. Each of these fractions was acid hy...

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“…suggested the possible involvement of endotoxinlike activity associated with those organisms (3,5,10,11,19). However, no precise chemical characterization was performed in these studies.…”

mentioning

confidence: 96%

Absence of lipopolysaccharide in the Lyme disease spirochete, Borrelia burgdorferi

Takayama¹,

Rothenberg²,

Barbour³

1987

Infect Immun

220

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“…Earlier work in our laboratory showed that the LPS extracted from T. hyodysenteriae is biologically active in its ability to act as a mitogen for splenocytes, to increase macrophage Fc and C3 receptor-mediated phagocytosis of red blood cells, to be cytotoxic for peritoneal cells in culture, and to generate a chemotaxin(s) in fresh serum (12). It is possible that LPS is liberated from T. hyodysenteriae during the disease state and that it is active in the induction of lesion formation.…”

Section: Resultsmentioning

confidence: 99%

Serotype-specific opsonization of Treponema hyodysenteriae

Nuessen¹,

Joens²

1982

Infect Immun

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Treponema hyodysenteriae was shown to attach to mouse peritoneal cells in the absence of serum opsonins in vitro. If serotype-specific antiserum from pigs was added to the media and treponemes of that corresponding serotype were employed in the assay, the amount of attachment increased an average of 3.7 times that of the control without pig sera. However, the amount of attachment was increased an average of only 1.5 times that of the control if organisms of any noncorresponding serotype of T. hyodysenteriae were used in the assay. Since the lipopolysaccharide (LPS) extracted from T. hyodysenteriae is the basis for serotyping the treponeme, the ability of these distinct LPS types to block attachment by blocking opsonization of the organisms was tested. Attachment, using corresponding antisera and treponemes, was blocked by LPS extracted from treponemes of that serotype, but not by LPS extracted from treponemes of other serotypes. These results indicate that antibody response to T. hyodysenteriae infection in pigs is serotype-specific. Furthermore, since opsonization and subsequent attachment of bacteria to phagocytes is known to be a protective mechanism, we suggest that the LPS may be an important antigen in the stimulation of host defense against the treponeme.

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“…They found that a phenol extract of Borrelia buccalis produces skin reactions and fever when injected into rabbits. Nuessen et al (21) reported that the LPS extracted from Treponema hyodysenteriae was capable of increasing of [3H]thymidine on murine splenocytes in vitro and was toxic for mouse peritoneal macrophages. Beck et al (3) and Isogai et al (14) also reported the mitogenic activity and adjuvanticity of leptospiral LPS.…”

Section: Discussionmentioning

confidence: 99%

Biological Activities of Lipopolysaccharide‐Like Substance (LLS) Extracted from Leptospira interrogans Serovar canicola Strain Moulton

Shimizu

Matsusaka²,

Nagakura³

et al. 1987

Microbiology and Immunology

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Biological activity of a lipopolysaccharide extracted from Treponema hyodysenteriae

Cited by 32 publications

References 18 publications

Absence of lipopolysaccharide in the Lyme disease spirochete, Borrelia burgdorferi

Absence of lipopolysaccharide in the Lyme disease spirochete, Borrelia burgdorferi

Serotype-specific opsonization of Treponema hyodysenteriae

Biological Activities of Lipopolysaccharide‐Like Substance (LLS) Extracted from Leptospira interrogans Serovar canicola Strain Moulton

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