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Infection and Immunity, May 2005, p. 3053-3062, Vol. 73, No. 5
0019-9567/05/$08.00+0     doi:10.1128/IAI.73.5.3053-3062.2005

Cytolethal Distending Toxin (CDT)-Negative Campylobacter jejuni Strains and Anti-CDT Neutralizing Antibodies Are Induced during Human Infection but Not during Colonization in Chickens

Manal AbuOun,1* Georgina Manning,1 Shaun A. Cawthraw,1 Anne Ridley,1 If H. Ahmed,1 Trudy M. Wassenaar,2 and Diane G. Newell1

Veterinary Laboratories Agency (Weybridge), Surrey, United Kingdom,1 Molecular Microbiology and Genomics Consultants, Zotzenheim, Germany2

Received 21 May 2004/ Returned for modification 3 June 2004/ Accepted 16 November 2004

The cytolethal distending toxin (CDT) of Campylobacter jejuni was detectable, using an in vitro assay, in most but not all of 24 strains tested. The reason for the absence of toxin activity in these naturally occurring CDT-negative C. jejuni strains was then investigated at the genetic level. CDT is encoded by three highly conserved genes, cdtA, -B, and -C. In the CDT-negative strains, two types of mutation were identified. The CDT activities of C. jejuni strains possessing both types of mutation were successfully complemented with the functional genes of C. jejuni 11168. The first type of mutation comprised a 667-bp deletion across cdtA and cdtB and considerable degeneration in the remainder of the cdt locus. Using a PCR technique to screen for this deletion, this mutation occurred in fewer than 3% of 147 human, veterinary, and environmental strains tested. The second type of mutation involved at least four nonsynonymous nucleotide changes, but only the replacement of proline with serine at CdtB position 95 was considered important for CDT activity. This was confirmed by site-directed mutagenesis. This type of mutation also occurred in fewer than 3% of strains as determined using a LightCycler biprobe assay. The detection of two CDT-negative clinical isolates raised questions about the role of CDT in some cases of human campylobacteriosis. To determine if anti-CDT antibodies are produced in human infection, a toxin neutralization assay was developed and validated using rabbit antisera. Pooled human sera from infected patients neutralized the toxin, indicating expression and immunogenicity during infection. However, no neutralizing antibodies were detected in colonized chickens despite the expression of CDT in the avian gut as indicated by reverse transcription-PCR.

* Corresponding author. Mailing address: Department of Food and Environmental Safety, Veterinary Laboratories Agency, New Haw, Addlestone, Surrey KT15 3NB, United Kingdom. Phone: 44 1932357738. Fax: 44 1932357595. E-mail: m.abuoun{at}vla.defra.gsi.gov.uk.

Editor: V. J. DiRita

Infection and Immunity, May 2005, p. 3053-3062, Vol. 73, No. 5
0019-9567/05/$08.00+0     doi:10.1128/IAI.73.5.3053-3062.2005




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