Production of a Monoclonal Antibody That Recognizes the Lipopolysaccharide of a <i>Campylobacter</i>‐Like Organism

Kuan, Steven; Coloe, Peter J.; Alderton, Malcolm R.

doi:10.1111/j.1348-0421.1992.tb02081.x

Cited by 3 publications

(3 citation statements)

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“…The electrophoretic mobility and antigenicity of C. hyoilei RMIT-32A LOS have previously been reported to be similar in size to C. coli with low molecular mass LOS antigens (Kuan et al, 1992; Mandatori & Penner, 1989). Whether C. hyoilei RMIT-32A LOS is similar to that of the LPS of a rough phenotype of Salmonella spp.…”

Section: Discussionmentioning

confidence: 83%

“…E. coli HB101 cells carrying the recombinant plasmids expressing the C. hyoilei antigens and C. hyoilei RMIT-32A cells were tested for the presence of LPS/LOS by treatment with 20 mg proteinase K for 16 h at 37 °C per 20 mg sample. Bacterial LPS and/or LOS was also extracted using a modified hot phenol procedure (Kuan et al, 1992).…”

Section: Methodsmentioning

confidence: 99%

“…In this paper we report on the molecular cloning and expression in E. coli of a DNA region encoding the synthesis of LOS antigen(s) from a newly identified species, Campylobacter hyoilei strain RMIT-32A (Alderton et al, 1995). C. hyoilei RMIT-32A had been shown to possess a low molecular mass glycolipid component (Kuan et al, 1992) which is likely to be LOS rather then traditional LPS characteristic of E. coli and Salmonella spp.…”

Section: Introductionmentioning

confidence: 99%

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Expression of Campylobacter hyoilei lipo-oligosaccharide (LOS) antigens in Escherichia coli

et al. 1997

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Campylobacter spp. are well recognized as primary pathogens in animals and in people. To isolate and define the genetic regions encoding major surface antigens of Campylobacter hyoilei, genomic DNA of the type strain of the species, RMIT-32A, was cloned into a cosmid vector, plA2917, in Escherichia coli and the resulting genomic library was screened using antiserum raised to the parent C. hyoilei strain. Six cosmid clones were found to express a series of immunoreactive bands in the 15-25 kDa range. These bands were proteinase Kresistant and were found in the LPS fraction of the cells, suggesting that the recombinant cosmids expressed C. hyoirei lipo-oligosaccharide (LOS) antigen(s). The minimum DNA insert size required for expression of C. hyoilei LOS antigen@) in E. coli was 11.8 kb. This region was subcloned into the plasmid vector pBR322. The partial sequencing of the 11.8 kb region showed that it contains two ORFs, designated rfbf and rfbP, showing homology with the rfbF gene from Serratia marcescens and the rfbP gene from Salmonella typhimurium. Both genes are involved in LPS synthesis. The region also contained a sequence homologous to the rfaC gene of E. coli and Sal. typhimurium which is involved in core oligosaccharide synthesis.

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Section: Discussionmentioning

confidence: 83%

Section: Methodsmentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

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Expression of Campylobacter hyoilei lipo-oligosaccharide (LOS) antigens in Escherichia coli

et al. 1997

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Multiplication of Orally AdministeredClostridium butyricumin Rats

Sato¹,

Tanaka²

1996

Microbial Ecology in Health and Disease

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A mouse monoclonal antibody (MAb) specific to the vegetative cell of Clostridium butyricurn strain MIYAIRI 588 (CBM588) was produced by immunisation with whole vegetative cells. This MAb (MAb-MS35) recognised a 35 KDa surface protein on CBM588 and did not cross-react with other intestinal bacteria including 16 strains of C. buiyricum. A close correlation was observed between CBM588 vegetative cell counts in rat faeces and enzyme-linked immunosorbent assay (ELISA) values using MAb-MS35. Consequently, this ELISA was used to specifically determine the concentrations of CBM588 in the faeces of rats. The aim of this study was to investigate the fate of CBM588 in the rat gastrointestinal tract after a single oral administration. CBM588 spores were administered intragastrically at lo7 cells per rat, and the faeces of each rat were collected twice daily. CBM588 vegetative cells were detected in the faeces for 3 d after administration. Total recovery of CBM588 vegetative cells from faeces were three to six times the spore dose. In addition, CBM588 vegetative cells were observed microscopically using faecal smears and an immunogold-silver staining method with MAb-MS35. These findings indicate that CBM588 spores germinate and the vegetative cells multiply in the gastrointestinal tract of rats.

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Multiplication of Orally AdministeredClostridium butyricumin Rats

Sato¹,

Tanaka²

1996

Microbial Ecology in Health & Disease

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Clostridium butyricum has been used as a probiotic in animals and humans for years, however, its fate in the intestine has not been clarified yet. We investigated the intestinal fate of C. butyricum using a selective medium and a monoclonal antibody after orally administering C. butyricum spores to rats. The number of C. butyricum, both viable and dead cells, in the intestinal contents were counted by enzyme-linked immunosorbent assay (ELISA) at various times after a single oral administration. The total viable number of C. butyricum was counted using a selective medium, and viable resting spores were selectively detected by treating the samples with ethanol. To investigate the intraluminal localization of the C. butyricum cells, frozen intestinal tracts were imprinted onto slides and stained with immunogold-silver. Total viable spores exceeded the number of viable resting spores by more than 10-fold from the proximal to middle of the small intestine 30 min after administration. Vegetative cells of C. butyricum were first detected in the dis-tal small intestine after 2 hr, and vegetative growth was observed from the cecum to the colon 5 hr after administration. Dead vegetative cells were detected 9 hr after administration, and C. butyricum cells were not detected in the intestine after 3 days. The C. butyricum cells in the intestinal imprints were stained specifically by immunogold-silver staining, and proliferative cells were observed in the cecum after 3 hr. These results suggest that the administered C. butyricum germinated in the upper small intestine, grew mainly from the distal small intestine to the colon and were excreted from the rat intestine within 3 days.

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Production of a Monoclonal Antibody That Recognizes the Lipopolysaccharide of a Campylobacter‐Like Organism

Cited by 3 publications

References 45 publications

Expression of Campylobacter hyoilei lipo-oligosaccharide (LOS) antigens in Escherichia coli

Expression of Campylobacter hyoilei lipo-oligosaccharide (LOS) antigens in Escherichia coli

Multiplication of Orally AdministeredClostridium butyricumin Rats

Multiplication of Orally AdministeredClostridium butyricumin Rats

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