Effect of normal and immune sera on Haemophilus ducreyi 35000HP and its isogenic MOMP and LOS mutants

Thomas J Hiltke; Margaret E Bauer; Julia  Klesney-Tait; Eric J Hansen; Robert S Munson Jr; Stanley M Spinola

doi:10.1006/mpat.1998.0250

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Effect of normal and immune sera on Haemophilus ducreyi 35000HP and its isogenic MOMP and LOS mutants

Journal article

Peer reviewed

Effect of normal and immune sera on Haemophilus ducreyi 35000HP and its isogenic MOMP and LOS mutants

Thomas J Hiltke, Margaret E Bauer, Julia Klesney-Tait, Eric J Hansen, Robert S Munson Jr and Stanley M Spinola

Microbial pathogenesis, Vol.26(2), pp.93-102

02/1999

DOI: 10.1006/mpat.1998.0250

PMID: 10090856

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Abstract

A bactericidal assay was developed in order to test the effect of hyperimmune rabbit sera on the viability of serum-resistant Haemophilus ducreyi 35000HP. Testing of several lots of rabbit complement and time course experiments showed that the serum-sensitive H. ducreyi CIPA77 was killed efficiently by 25% complement at 35 degrees C in 3 h. We hypothesized that incubation of 35000HP under these conditions with the appropriate bactericidal antibody would kill this strain. A panel of high titre rabbit antisera was developed and tested against 35000HP. The panel included antisera raised to whole cells, total membranes, Sarkosyl-insoluble outer membrane proteins, the H. ducreyi lipoprotein, and the peptidoglycan-associated lipoprotein. None of the antisera convincingly showed bactericidal activity. The bactericidal assay was also used to determine the effect of normal human serum (NHS) on isogenic mutants of 35000HP. 35000HP-RSM2, an Omegakan insertion mutant that expresses a truncated lipooligosaccharide, was as resistant to NHS as its parent. A mutant deficient in expression of the major outer membrane protein (35000. 60) was sensitive to NHS. We conclude that 35000HP is relatively resistant to normal and hyperimmune sera, and that the major outer membrane protein contributes to this resistance.

Mutation

Chancroid - immunology

Rabbits

Bacterial Outer Membrane Proteins - immunology

Enzyme-Linked Immunosorbent Assay

Electrophoresis, Polyacrylamide Gel

Humans

Antigens, Bacterial - immunology

Complement System Proteins - immunology

Haemophilus ducreyi - genetics

Haemophilus ducreyi - immunology

Blood Bactericidal Activity - immunology

Immunization - veterinary

Antibodies, Monoclonal

Colony Count, Microbial

Haemophilus ducreyi - growth & development

Antigens, Bacterial - biosynthesis

Lipopolysaccharides - immunology

Blotting, Western

Animals

Bacterial Outer Membrane Proteins - genetics

Details

Title: Subtitle: Effect of normal and immune sera on Haemophilus ducreyi 35000HP and its isogenic MOMP and LOS mutants
Creators: Thomas J Hiltke - School of Medicine, Indiana University, Indianapolis, Indiana 46202, USA
Margaret E Bauer
Julia Klesney-Tait
Eric J Hansen
Robert S Munson Jr
Stanley M Spinola
Resource Type: Journal article
Publication Details: Microbial pathogenesis, Vol.26(2), pp.93-102
DOI: 10.1006/mpat.1998.0250
PMID: 10090856
ISSN: 0882-4010
eISSN: 1096-1208
Grant note: AI09971 / NIAID NIH HHS F32 AI009971-01 / NIAID NIH HHS AI31494 / NIAID NIH HHS F32 AI009971-02 / NIAID NIH HHS AI27863 / NIAID NIH HHS F32 AI009971-03 / NIAID NIH HHS F32 AI009971 / NIAID NIH HHS
Language: English
Date published: 02/1999
Academic Unit: Pulmonary, Critical Care, and Occupational Medicine; Internal Medicine
Record Identifier: 9984094645802771

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