Monoclonal antibodies reactive with all strains of Haemophilus ducreyi

Hansen, Eric J.; Loftus, Tiffany

doi:10.1128/iai.44.1.196-198.1984

Cited by 32 publications

(9 citation statements)

References 17 publications

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“…MAbs. Lymphocyte hybridomas secreting monoclonal antibodies (MAbs) reactive with H. ducreyi cell envelope antigens were obtained from hybridoma fusions involving the use of splenocytes from mice immunized with H. ducreyi 35000 cell envelopes (15). MAb 1F8, reactive with the 49-kDa cell envelope antigen, and MAb 2H4, directed against the 23-kDa cell envelope antigen, were identified by their reactivities in Western immunoblot analysis with H. ducreyi cell envelopes as antigens (14a).…”

Section: Bacteriophage X1105mentioning

confidence: 99%

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Use of electroporation to construct isogenic mutants of Haemophilus ducreyi

et al. 1992

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Little is known about the genetics of Haemophilus ducreyi, the etiologic agent of chancroid. To develop a method for constructing isogenic mutants of this organism that could be utilized in pathogenesis-related studies, electroporation techniques were evaluated as a means of introducing DNA into this organism. Electroporation of the plasmid shuttle vector pLS88 into H. ducreyi yielded approximately 10(6) antibiotic-resistant transformants per microgram of plasmid DNA. Studies of the feasibility of moving mutated genes into H. ducreyi were initiated by using NotI linker insertion and mini-Tn10kan mutagenesis techniques to introduce insertion mutations into cloned H. ducreyi genes encoding cell envelope antigens. In the former case, a gene encoding chloramphenicol acetyltransferase was then inserted into the NotI linker site created in the cloned H. ducreyi gene. The recombinant Escherichia coli strains containing these mutated plasmids no longer expressed the homologous H. ducreyi cell envelope antigens, as evidenced by their lack of reactivity with monoclonal antibody probes for these H. ducreyi proteins. Subsequent electroporation of both circular and linearized forms of plasmids carrying these mutated H. ducreyi genes into the homologous wild-type strain of H. ducreyi yielded antibiotic-resistant transformants which also lacked reactivity with the cell envelope antigen-specific monoclonal antibodies. Southern blot analysis confirmed that homologous recombination had occurred in these monoclonal antibody-unreactive transformants, resulting in the replacement of the wild-type allele with the mutated allele. Allelic exchange was most efficient when linear DNA molecules were used for electroporation. These results indicate that electroporation methods can be utilized to construct isogenic mutants of H. ducreyi.

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Section: Bacteriophage X1105mentioning

confidence: 99%

“…The colony blot radioimmunoassay procedure was used to screen bacterial colonies for their reactivities with H. ducreyi-specific MAbs (13,15).…”

Section: Bacteriophage X1105mentioning

confidence: 99%

Use of electroporation to construct isogenic mutants of Haemophilus ducreyi

et al. 1992

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“…Complex whorls (A); parallel chains (B); streptobacillary chains (C); short chains and individual bacilli (D). Kindly provided by D. Hardy (49,120…”

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Biology of Haemophilus ducreyi.

Albritton¹

1989

Microbiological Reviews

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“…. 90-99 L-Aspartate, L-lysyl-alanine, L-lysyl-L-lysine.................................................................................................................. 80-89 Glycyl-L-phenylalanine, glycyl-L-proline ..................................................................................................................... 70-79 L-Leucyl-L-alanine, L-seryl-L-methionine, L-leucyl-glycine, L-ornithine .............................................................................. [60][61][62][63][64][65][66][67][68][69].…”

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Chancroid and Haemophilus ducreyi.

Morse¹

1989

Clinical Microbiology Reviews

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The organism was observed both within and outside of neutrophils. It grew in chains, especially in tissue, but also appeared as single cells in secretions. Ducrey consistently observed this organism from a cleansed soft chancre, in pus from an unruptured bubo, and in the deep portion of an abscess. Since he was unable to culture the organism in vitro, he used serial cutaneous inoculations to establish the specificity of the infectious agent. His observations were confirmed by Krefting in 1892 and also by Unna in 1892 (as cited in reference 160). There is some controversy concerning the first culture isolation of Haemophilus ducreyi. Sullivan (160) attributed the first cultures of the bacillus of Ducrey to Lenglet in 1898 and Bezancon, Griffin, and LeSourd in 1900. However, Istamanoff (131) and Akspianz (38) have been credited with earlier isolations.

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Monoclonal antibodies reactive with all strains of Haemophilus ducreyi

Cited by 32 publications

References 17 publications

Use of electroporation to construct isogenic mutants of Haemophilus ducreyi

Use of electroporation to construct isogenic mutants of Haemophilus ducreyi

Biology of Haemophilus ducreyi.

Chancroid and Haemophilus ducreyi.

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