Production and characterization of monoclonal antibodies specific for lipooligosaccharide of Serpulina hyodysenteriae

Westerman, R B; Phillips, R. E.; Joens, Lynn A.

doi:10.1128/jcm.33.8.2145-2149.1995

Cited by 8 publications

(6 citation statements)

References 23 publications

Supporting

Mentioning

Contrasting

Order By: Relevance

“…16 Due to strong crossreactions between the different species of Serpulina, development of immunohistochemical methods based on monoclonal antibodies for specific detection of spirochetes in situ is difficult. 22,43 By the FISH technique, even single cells present in or on the mucosa can be Diarrhea in Growing Pigs detected in formalin-fixed tissue samples used for histology. This study shows the applicability of the method for identification of B. pilosicoli in pathogenesis studies.…”

Section: Discussionmentioning

confidence: 99%

Scanning Electron Microscopy and Fluorescent In Situ Hybridization of Experimental Brachyspira (Serpulina) pilosicoli Infection in Growing Pigs

Jensen¹,

Möller²,

Boye³

et al. 2000

Vet Pathol

View full text Add to dashboard Cite

Two groups of six 8-week-old pigs were challenged with 1x10(9) cfu Brachyspira (Serpulina) pilosicoli or Serpulina intermedia daily for 3 consecutive days to study the pathology of porcine colonic spirochetosis by scanning electron microscopy (SEM) and fluorescent in situ hybridization (FISH) with oligonucleotide probes targeting ribosomal RNA specific for B. pilosicoli and the genus Brachyspira/Serpulina. Six pigs served as noninoculated controls. The animals were euthanatized successively between postinoculation days 14 and 24. B. pilosicoli was reisolated in feces from all of the inoculated pigs; however, only two pigs developed transient watery diarrhea. S. intermedia was reisolated from four of the inoculated pigs, but clinical signs were not observed. Gross examination of the B. pilosicoli-infected pigs revealed dilated large intestines with a hyperemic mucosa, whereas the large intestines of the S. intermedia-inoculated pigs and the control pigs appeared normal. SEM examination of B. pilosicoli-infected pigs revealed degenerated epithelial cells and spirochetal colonization of the colonic mucosa in four pigs. By FISH, B. pilosicoli cells were found colonizing and invading the surface epithelium and the crypts in all the pigs. Spirochetal crypt colonization markedly exceeded the occurrence of spirochetes on the mucosal surface. SEM examination of S. intermedia-inoculated pigs revealed no abnormalities, and Serpulina cells were detected only sporadically in the otherwise normal-appearing mucosa of four pigs by FISH. The results provide further evidence that B. pilosicoli is associated with colitis in pigs, although the gross lesions are mild. The spirochete is capable of colonizing the large intestine, inducing mucosal damage, invasion of the crypt and surface epithelium, and focal infiltration of the lamina propria. In addition, the study shows the applicability of FISH for specific identification of B. pilosicoli in formalin-fixed tissue.

show abstract

Section: Discussionmentioning

confidence: 99%

Scanning Electron Microscopy and Fluorescent In Situ Hybridization of Experimental Brachyspira (Serpulina) pilosicoli Infection in Growing Pigs

Jensen¹,

Möller²,

Boye³

et al. 2000

Vet Pathol

View full text Add to dashboard Cite

show abstract

“…A mouse MAb was produced by using whole-cell suspensions of killed S. hyodysenteriae B234 as previously described (36). Supernatants from hybridomas were screened for antibody production against whole-cell lysates of S. hyodysenteriae B234 and S. innocens B256 by an enzyme-linked immunosorbent assay (36). One hybridoma secreting monoclonal immunoglobulin G antibodies reacted with S. hyodysenteriae B234 and S. innocens B256.…”

Section: Methodsmentioning

confidence: 99%

“…One hybridoma secreting monoclonal immunoglobulin G antibodies reacted with S. hyodysenteriae B234 and S. innocens B256. The MAb, designated 7G2, produced a banding pattern by immunoblot against whole-cell lysate and purified PF (35,36) that was suggestive of reactivity with the 37-, 34-, and 32-kDa FlaB proteins of S. hyodysenteriae B204.…”

Section: Methodsmentioning

confidence: 99%

Immunoblot reactivity of polyclonal and monoclonal antibodies with periplasmic flagellar proteins FlaA1 and FlaB of porcine Serpulina species

Fisher

Duhamel

Westerman

et al. 1997

Clin Diagn Lab Immunol

Self Cite

View full text Add to dashboard Cite

The periplasmic-flagellum (PF) proteins of Triton X-100-soluble and Triton X-100-insoluble sodium dodecyl sulfate-treated fractions from reference and field strains of Serpulina hyodysenteriae, Serpulina innocens, and Serpulina pilosicoli were characterized by Western blotting with a rabbit polyclonal antibody (PAb) specific for the 44-kDa PF sheath protein of S. hyodysenteriae (Z. Li, F. Dumas, D. Dubreuil, and M. Jacques, J. Bacteriol. 175:8000-8007, 1993) and a murine monoclonal antibody (MAb), designated 7G2, specific for the PF core FlaB proteins of S. hyodysenteriae. The MAb 7G2 reacted with a conserved epitope present in the 37-, 34-, and 32-kDa PF core FlaB proteins of all Serpulina species. This suggested that the core FlaB proteins are conserved among porcine Serpulina species. An immunoreactive band of approximately 44 kDa was present with all S. hyodysenteriae, S. innocens, and S. pilosicoli strains that were reacted with the PAb. The specificities of the PAb and the MAb for the FlaA1 and FlaB proteins of Serpulina species were confirmed by N-terminal amino acid sequencing of 44-and 37-kDa proteins, respectively, of S. hyodysenteriae and S. pilosicoli. Results from this study provide further evidence that the 44-kDa protein FlaA1 and the 37-, 34-, and 32-kDa FlaB proteins are conserved among porcine Serpulina species.

show abstract

“…Transmission electron microscopy (TEM) may give some indicative information regarding the species involved, but pathogenic and non-pathogenic spirochaetes cannot be differentiated by TEM. Furthermore, the development of immunohistochemical methods for specific detection of S. hyodysenteriue is difficult due to strong cross-reactions between the different species of Serpulinu (16,18,28).…”

mentioning

confidence: 99%

Association of Serpulina hyodysenteriae with the colonic mucosa in experimental swine dysentery studied by fluorescent in situ hybridization

Jensen¹,

Boye²,

Möller³

et al. 1998

APMIS

View full text Add to dashboard Cite

Association of Serpulina hyodysenteriae with the colonic mucosa in experimental swine dysentery studied by fluorescent in situ hybridization.The localization of Serpulina hyodysenteriae in experimental swine dysentery was studied by fluorescent in situ hybridization (FISH) using an oligonucleotide probe targeting the 23s rRNA of S. hyodysenteriae. Nine 8-week-old pigs were challenged. Seven of the pigs were intragastrically dosed with 1 X lo9 cfu S. hyodysenteriae for 3 consecutive days, whereas two pigs were infected by contact. Six non-challenged pigs served as negative controls. The challenged pigs developed clinical swine dysentery from 8 to 14 days postinfection with typical gross lesions. By FISH S. hyodysenteriae cells in huge numbers were found colonizing the mucus layer, the luminal epithelium and the crypts of the large intestinal mucosa. The orientation of the spirochaetes to the epithelium appeared to be random. Spirochaetes in clusters or as single cells were invading the epithelium and were occasionally detected in the adjacent lamina propria. The distribution of spirochaetes in the mucosa provides further evidence that S. hyodysenteriae is intimately associated with the mucus layer and the epithelium in a random pattern. Furthermore, the results demonstrate the applicability of FISH for specific detection of S. hyodysenteriae cells in clusters or as single cells in formalin-fixed tissue samples.

show abstract

Production and characterization of monoclonal antibodies specific for lipooligosaccharide of Serpulina hyodysenteriae

Cited by 8 publications

References 23 publications

Scanning Electron Microscopy and Fluorescent In Situ Hybridization of Experimental Brachyspira (Serpulina) pilosicoli Infection in Growing Pigs

Scanning Electron Microscopy and Fluorescent In Situ Hybridization of Experimental Brachyspira (Serpulina) pilosicoli Infection in Growing Pigs

Immunoblot reactivity of polyclonal and monoclonal antibodies with periplasmic flagellar proteins FlaA1 and FlaB of porcine Serpulina species

Association of Serpulina hyodysenteriae with the colonic mucosa in experimental swine dysentery studied by fluorescent in situ hybridization

Contact Info

Product

Resources

About