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Formaldehyde treatment increases the immunogenicity and decreases the toxicity of Haemophilus ducreyi cytolethal distending toxin.

Journal article
Authors Teresa Lagergård
Annika Lundqvist
Catharina Wising
Vivianne Gabrielsson
Karin Ahlman
Published in Vaccine
Volume 25
Issue 18
Pages 3606-14
ISSN 0264-410X
Publication year 2007
Published at Institute of Biomedicine, Department of Microbiology and Immunology
Pages 3606-14
Language en
Links dx.doi.org/10.1016/j.vaccine.2007.0...
Keywords Animals, Antibodies, Bacterial, blood, Bacterial Toxins, administration & dosage, immunology, isolation & purification, toxicity, Chancroid, prevention & control, Formaldehyde, pharmacology, Haemophilus Vaccines, Haemophilus ducreyi, growth & development, immunology, Hela Cells, Humans, Immunization, Mice, Mice, Inbred BALB C, Neutralization Tests, Toxoids, administration & dosage, immunology
Subject categories Medical and Health Sciences

Abstract

Haemophilus ducreyi cytolethal distending toxin (HdCDT) is a tripartite AB toxin, which causes DNA damage in affected cells. We investigated the effects of formaldehyde on the chemical, biological, and immunological properties of the HdCDT complex, which was purified by immobilizing the glutathione S-transferase (GST)-CdtB fusion protein, followed by binding of the CdtA and CdtC recombinant proteins. The HdCDT was treated with increasing concentrations of formaldehyde in the presence of lysine. The treatment of HdCDT at 1 and 0.1 mg protein/ml with 320 and 80 mM of formaldehyde, respectively, resulted in the complete abrogation of cytotoxic activity, loss of DNase activity, and loss of binding capacity to HeLa cells. The toxoid showed protein bands of 75-150 kDa in SDS-PAGE, composed of the three cross-linked CDT components detected by immunoblotting. Three doses of 10 microg protein/mouse of the formaldehyde-treated HdCDT elicited toxin-neutralizing antibodies at titers about 200 times higher than those elicited by the native toxin. The described methodology may be applied to produce immunogenic toxoids from other CDTs, which might be used as candidate components in vaccines against CDT-producing bacteria, including H. ducreyi.

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