Cloning and expression of the 75 kDa DnaK-like protein of Chlamydia psittaci and the evaluation of the recombinant protein by immunoblotting and indirect ELISA

Anderson, I.E.; Craggs, Joanna K.; Dunbar, Susanna M.; Herring, A. J.

doi:10.1016/s0378-1135(97)00166-1

Cited by 2 publications

(3 citation statements)

References 28 publications

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“…In this study, we evaluated the protective immunity induced by the dnaK gene of C. abortus. Indeed, the bacterial heat shock proteins and more particularly, the DnaK protein (Hsp70) of C. abortus are known to be strong immunogens [1,34]. DnaK is located in the outer membrane of C. trachomatis and is accessible to the monoclonal antibody [21].…”

Section: Discussionmentioning

confidence: 99%

“…Proteins of the heat shock protein (Hsp) family are evaluated as good candidates for this approach of vaccination against C. abortus [1]. Widely distributed in nature, they are implicated in the folding, unfolding and translocation of proteins and also in the assembly and disassembly of protein complexes as immunoglobulins (Ig), T-cell receptors (TCR) and major histocompatibility complexes (MHC) [34].…”

Section: Introductionmentioning

confidence: 99%

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Protection evaluation against Chlamydophila abortus challenge by DNA vaccination with a dnaK-encoding plasmid in pregnant and non-pregnant mice

Héchard¹,

Grépinet²,

Rodolakis³

2002

Vet. Res.

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-Mice were intramuscularly immunized with a dnaK-encoding DNA plasmid. The protective effect of DNA immunization against Chlamydophila abortus infection was studied in pregnant and non-pregnant mice models. In non-pregnant mice, the dnaK vaccine induced a specific humoral response with the predominant IgG2a isotype, which failed to have in vitro neutralizing properties. No delayed-type hypersensitivity reaction was observed and the spleens of dnaK vaccinated-mice were not protected against C. abortus challenge. In pregnant mice, the dnaK vaccine induced a non-specific partial protection from abortion. This may be due to the immunogenic properties of the CpG motifs of bacterial DNA present in the vaccinal plasmid backbone. Nevertheless, spleens of dnaK vaccinated-pregnant mice were not protected. DNA immunization / Chlamydophila abortus / DnaK / Hsp70Résumé -Évaluation de la protection induite par vaccination ADN avec un plasmide codant pour dnaK contre une épreuve par Chlamydophila abortus chez des souris gestantes ou nongestantes. Des souris ont été immunisées par l'injection intramusculaire d'un plasmide codant pour la protéine DnaK. L'effet protecteur de la vaccination ADN contre une infection par Chlamydophila abortus a été évalué dans des modèles de souris gestantes ou non-gestantes. La vaccination ADN avec le gène dnaK induit une réponse humorale spécifique chez les souris non-gestantes avec une prédo-minance d'IgG2a qui n'ont pas présenté d'activité neutralisante in vitro. Aucune réaction d'hypersensibilité retardée n'a été observée et les rates des souris vaccinées avec le vaccin ADN-dnaK n'ont pas été protégées contre l'infection par C. abortus. Chez les souris gestantes, le vaccin ADN-dnaK ré-duit partiellement et de façon non spécifique les avortements. Les îlots CpG de l'ADN bactérien, connus pour leur propriétés immunogéniques, pourraient être responsables de cet effet protecteur non spécifique. Cependant, aucune élimination des bactéries n'a été observée dans les rates des souris gestantes.vaccination ADN / Chlamydophila abortus / DnaK / Hsp70 313 Vet. Res. 33 (2002) 313-326

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Section: Discussionmentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

Protection evaluation against Chlamydophila abortus challenge by DNA vaccination with a dnaK-encoding plasmid in pregnant and non-pregnant mice

Héchard¹,

Grépinet²,

Rodolakis³

2002

Vet. Res.

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“…the interest of microbiologists for many years, since they represent major targets of the host's immune response (19). Although less extensively studied than GroEL, DnaK homologues of many bacterial pathogens have been found to be immunogenic in humans or animals (1,2,5,8,10,47). Furthermore, as has been shown by the experimental infection of mice with Borrelia burgdorferi, immunization with proteins containing DnaK-specific sequences may protect against microbial infection (4).…”

mentioning

confidence: 99%

Cloning and Expression of the dnaK Gene of Campylobacter jejuni and Antigenicity of Heat Shock Protein 70

Thiès¹,

Karch

Hartung³

et al. 1999

Infect Immun

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Campylobacter jejuni is a leading cause of infectious diarrhea throughout the world. In addition, there is growing evidence that Guillain-Barré syndrome, an inflammatory demyelinating disease of the peripheral nervous system, is frequently preceded byC. jejuni infection. In the present study, thehrcA-grpE-dnaK gene cluster of C. jejuni was cloned and sequenced. The dnaK gene consists of an open reading frame of 1,869 bp and encodes a protein with a high degree of homology to other bacterial 70-kDa heat shock proteins (HSPs). The overall percentages of identity to the HSP70 proteins ofHelicobacter pylori, Borrelia burgdorferi,Chlamydia trachomatis, and Bacillus subtiliswere calculated to be 78.1, 60.5, 57.2, and 53.8%, respectively. Regions similar to the Escherichia coli ς70promoter consensus sequence and to a cis-acting regulatory element (CIRCE) are located upstream of the hrcA gene. Following heat shock, a rapid increase of dnaK mRNA was detectable, which reached its maximum after 20 to 30 min. A 6-His-tagged recombinant DnaK protein (rCjDnaK-His) was generated inE. coli, after cloning of the dnaK coding region into pET-22b(+), and purified by affinity and gel filtration chromatography. Antibody responses to rCjDnaK-His were significantly elevated, compared to those of healthy individuals, in about one-third of the serum specimens obtained from C. jejuni enteritis patients.

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Cloning and expression of the 75 kDa DnaK-like protein of Chlamydia psittaci and the evaluation of the recombinant protein by immunoblotting and indirect ELISA

Cited by 2 publications

References 28 publications

Protection evaluation against Chlamydophila abortus challenge by DNA vaccination with a dnaK-encoding plasmid in pregnant and non-pregnant mice

Protection evaluation against Chlamydophila abortus challenge by DNA vaccination with a dnaK-encoding plasmid in pregnant and non-pregnant mice

Cloning and Expression of the dnaK Gene of Campylobacter jejuni and Antigenicity of Heat Shock Protein 70

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