Immunization with Recombinant Corynebacterium pseudotuberculosis Heat-Shock Protein (Hsp)-60 is Able to Induce an Immune Response in Mice, But Fails to Confer Protection Against Infection

Pinho, J. M. R.; Dorella, Fernanda Alves; Coelho, Keila da Silva; Fonseca, Cristina Toscano; Cardoso, Fernanda C.; Meyer, Roberto; Portela, Ricardo Wagner; Oliveira, Sérgio C.; Miyoshi, Anderson; Azevedo, Vasco

doi:10.2174/1874318800903010022

Cited by 3 publications

(4 citation statements)

References 15 publications

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“…The hsp60 DNA vaccine induced a cellular immune response but failed to confer protective immunity to the host, which corroborates a previous study that found that an Hsp60 protein subunit vaccine also did not confer protection [13]. …”

Section: Discussionsupporting

confidence: 89%

“…The Hsp60 protein of C. pseudotuberculosis , using a protein subunit as immunogen against CLA, failed to confer protection against infection with C. pseudotuberculosis in mice [13]. Using an alternative strategy, we assessed the feasibility of using DNA encoding hsp60 for protection against experimental challenge with C. pseudotuberculosis .…”

Section: Resultsmentioning

confidence: 99%

“…To examine the functional activity of the C. pseudotuberculosis hsp60 gene, E. coli B178 groEL44 mutant (Δ groEL44 ) was transformed with a pProEx-Hta/ hsp60 plasmid constructed by Pinho et al [13]. The E. coli Δ groEL44 mutant is a strain that bears a temperature-sensitive allele of groEL , namely groEL44 .…”

Section: Methodsmentioning

confidence: 99%

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Molecular characterization of the Corynebacterium pseudotuberculosis hsp60-hsp10 operon, and evaluation of the immune response and protective efficacy induced by hsp60 DNA vaccination in mice

et al. 2011

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BackgroundHeat shock proteins (HSPs) are important candidates for the development of vaccines because they are usually able to promote both humoral and cellular immune responses in mammals. We identified and characterized the hsp60-hsp10 bicistronic operon of the animal pathogen Corynebacterium pseudotuberculosis, a Gram-positive bacterium of the class Actinobacteria, which causes caseous lymphadenitis (CLA) in small ruminants.FindingsTo construct the DNA vaccine, the hsp60 gene of C. pseudotuberculosis was cloned in a mammalian expression vector. BALB/c mice were immunized by intramuscular injection with the recombinant plasmid (pVAX1/hsp60).ConclusionThis vaccination induced significant anti-hsp60 IgG, IgG1 and IgG2a isotype production. However, immunization with this DNA vaccine did not confer protective immunity.

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Section: Discussionsupporting

confidence: 89%

Section: Resultsmentioning

confidence: 99%

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Molecular characterization of the Corynebacterium pseudotuberculosis hsp60-hsp10 operon, and evaluation of the immune response and protective efficacy induced by hsp60 DNA vaccination in mice

et al. 2011

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“…This study was performed because of the need for a live vaccine to prevent caseous lymphadenitis. To date, the mutant strains that have been evaluated as possible C. pseudotuberculosis immunogens have failed to confer adequate protection to susceptible animals 9,21,22 . Here, the aroA mutant of C. pseudotuberculosis was chosen based on the good results that were obtained by applying the aroA mutant of Salmonella enterica, subsp.…”

Section: Cytokine Measurement Induced By Vaccinationmentioning

confidence: 99%

Evaluation of the aroA mutant of Corynebacterium pseudotuberculosis in cellular and murine models

et al. 2016

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Caseous lymphadenitis of small ruminants causes economic losses worldwide. To date, no effective vaccine has been developed against the causative agent of this disease, Corynebacterium pseudotuberculosis. The objective of the present work was to evaluate an aroA mutant gene strain of C. pseudotuberculosis in cellular and murine models, for attenuation and the ability to stimulate an immune response. The intracellular survival of the aroA mutant strain and the wild type strain (WT) of C. pseudotuberculosis was evaluated in J774A.1 murine macrophages using a multiplicity of infection (MOI) of 1:1 with the following infection times: 30 min, and 1, 2, 4, 8, 12, and 24 h. The largest difference in the intracellular survival of the mutant was observed 30 min post-infection. After subcutaneous skin vaccination, the subcutaneous lesion progression observed on the 14 th day was more severe in those animals that were vaccinated with the WT strain. An analysis of the residual virulence in the murine model did not reveal any bacteria in mice vaccinated with the aroA strain on day 28 post-vaccination. Mice vaccinated with the mutant showed 50 % protection against the intraperitoneal challenge, exceeding that of the control group (41.67 %). We conclude that the virulence of the aroA mutant was significantly attenuated in both cellular and murine models according to the residual virulence detected in mice. However, vaccination with the mutant failed to confer at least 80 % protection, which is desirable for an immunogen. Hence, this study contributes to the knowledge of the immune response against Corynebacterium pseudotuberculosis.

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Ion Torrent‐based transcriptional assessment of a Corynebacterium pseudotuberculosis equi strain reveals denaturing high‐performance liquid chromatography a promising rRNA depletion method

Castro

Seyffert

Ramos

et al. 2013

Microbial Biotechnology

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Corynebacterium pseudotuberculosis equi is a Gram-positive pathogenic bacterium which affects a variety of hosts. Besides the great economic losses it causes to horse-breeders, this organism is also known to be an important infectious agent to cattle and buffaloes. As an outcome of the efforts in characterizing the molecular basis of its virulence, several complete genome sequences were made available in recent years, enabling the large-scale assessment of genes throughout distinct isolates. Meanwhile, the RNA-seq stood out as the technology of choice for comprehensive transcriptome studies, which may bring valuable information regarding active genomic regions, despite of the still impeditive associated costs. In an attempt to increase the use of generated reads per instrument run, by effectively eliminating unwanted rRNAs from total RNA samples without relying on any commercially available kits, we applied denaturing high-performance liquid chromatography (DHPLC) as an alternative method to assess the transcriptional profile of C. pseudotuberculosis. We have found that the DHPLC depletion method, allied to Ion Torrent sequencing, allows mapping of transcripts in a comprehensive way and identifying novel transcripts when a de novo approach is used. These data encourage us to use DHPLC in future transcriptional evaluations in C. pseudotuberculosis.

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Immunization with Recombinant Corynebacterium pseudotuberculosis Heat-Shock Protein (Hsp)-60 is Able to Induce an Immune Response in Mice, But Fails to Confer Protection Against Infection

Cited by 3 publications

References 15 publications

Molecular characterization of the Corynebacterium pseudotuberculosis hsp60-hsp10 operon, and evaluation of the immune response and protective efficacy induced by hsp60 DNA vaccination in mice

Molecular characterization of the Corynebacterium pseudotuberculosis hsp60-hsp10 operon, and evaluation of the immune response and protective efficacy induced by hsp60 DNA vaccination in mice

Evaluation of the aroA mutant of Corynebacterium pseudotuberculosis in cellular and murine models

Ion Torrent‐based transcriptional assessment of a Corynebacterium pseudotuberculosis equi strain reveals denaturing high‐performance liquid chromatography a promising rRNA depletion method

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