Mycobacterium avium subsp. paratuberculosis-specific mpt operon expressed in M. bovis BCG as vaccine candidate

Heinzmann, Julia; Wilkens, Mirja R.; Dohmann, Karen; Gerlach, Gerald-F.

doi:10.1016/j.vetmic.2008.01.014

Cited by 11 publications

(9 citation statements)

References 29 publications

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“…Several iron related genes were downregulated in tissues including LSP15, a MAP unique pathogenicity island that encodes a ferric uptake regulator (MAP3773c), as well as the ABC transporter (MAP3731c - MAP3736c), and a possible siderophore biosynthesis operon (MAP3741-MAP3746) that contains a FUR binding box within the intergenic region [42]. This is of significant interest as part of this region (MAP3731c-MAP3736c) has previously been shown to be immunogenic and preliminary studies indicate its use as a potential vaccine candidate[43]. Furthermore, our genome analysis revealed that a type VII secretory system (esx3) was located immediately downstream of LSP15.…”

Section: Discussionmentioning

confidence: 99%

Primary transcriptomes of Mycobacterium avium subsp. paratuberculosis reveal proprietary pathways in tissue and macrophages

et al. 2010

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BackgroundMycobacterium avium subsp. paratuberculosis (MAP) persistently infects intestines and mesenteric lymph nodes leading to a prolonged subclinical disease. The MAP genome sequence was published in 2005, yet its transcriptional organization in natural infection is unknown. While prior research analyzed regulated gene sets utilizing defined, in vitro stress related or advanced surgical methods with various animal species, we investigated the intracellular lifestyle of MAP in the intestines and lymph nodes to understand the MAP pathways that function to govern this persistence.ResultsOur transcriptional analysis shows that 21%, 8% and 3% of the entire MAP genome was represented either inside tissues, macrophages or both, respectively. Transcripts belonging to latency and cell envelope biogenesis were upregulated in the intestinal tissues whereas those belonging to intracellular trafficking and secretion were upregulated inside the macrophages. Transcriptomes of natural infection and in vitro macrophage infection shared genes involved in transcription and inorganic ion transport and metabolism. MAP specific genes within large sequence polymorphisms of ancestral M. avium complex were downregulated exclusively in natural infection.ConclusionsWe have unveiled common and unique MAP pathways associated with persistence, cell wall biogenesis and virulence in naturally infected cow intestines, lymph nodes and in vitro infected macrophages. This dichotomy also suggests that in vitro macrophage models may be insufficient in providing accurate information on the events that transpire during natural infection. This is the first report to examine the primary transcriptome of MAP at the local infection site (i.e. intestinal tissue). Regulatory pathways that govern the lifecycle of MAP appear to be specified by tissue and cell type. While tissues show a "shut-down" of major MAP metabolic genes, infected macrophages upregulate several MAP specific genes along with a putative pathogenicity island responsible for iron acquisition. Many of these regulatory pathways rely on the advanced interplay of host and pathogen and in order to decipher their message, an interactome must be established using a systems biology approach. Identified MAP pathways place current research into direct alignment in meeting the future challenge of creating a MAP-host interactome.

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Section: Discussionmentioning

confidence: 99%

Primary transcriptomes of Mycobacterium avium subsp. paratuberculosis reveal proprietary pathways in tissue and macrophages

et al. 2010

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“…Recombinant MAP vaccines should have various merits over killed or attenuated vaccines in terms of antigen production and human safety. The most commonly evaluated recombinant proteins have been Hsp 70 (Koets et al, 2006), antigen 85, 74F, SOD, 35 kDa (Chen et al, 2008; Kathaperumal et al, 2008; Park et al, 2008), mpt (Heinzmann et al, 2008), 95 kDa (Bull et al, 2007), P22 (Rigden et al, 2006), 65 kDa (Velaz-Faircloth et al, 1999), and 16.8 kDa (Kadam et al, 2009). Many of the recombinant vaccines were reported to induce strong cellular as well as antibody mediated immune responses (Rigden et al, 2006; Kathaperumal et al, 2008; Roupie et al, 2008).…”

Section: Introductionmentioning

confidence: 99%

Cellular and humoral immune responses in sheep vaccinated with candidate antigens MAP2698c and MAP3567 from Mycobacterium avium subspecies paratuberculosis

Gurung

Purdie

Whittington

et al. 2014

Front. Cell. Infect. Microbiol.

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Control of Johne's disease, caused by Mycobacterium avium subspecies paratuberculosis (MAP) in ruminants using commercially available vaccine reduces production losses, mortality, fecal shedding and histopathological lesions but does not provide complete protection from infection and interferes with serological diagnosis of Johne's disease and bovine tuberculosis. At this time no recombinant antigens have been found to provide superior protection compared to whole killed or live-attenuated MAP vaccines. Therefore, there is a need to evaluate more candidate MAP antigens. In this study recombinant MAP antigens MAP2698c and MAP3567 were formulated with four different MONTANIDE™ (ISA 50V2, 61VG, 71VG, and 201VG) adjuvants and evaluated for their ability to produce specific immune responses in vaccinated sheep. The cellular immune response was measured with an interferon-gamma (IFN-γ) release assay and the humoral immune response was measured by antibody detection enzyme linked immunosorbent assay. Recombinant vaccine formulation with the antigen MAP2698c and MONTANIDE™ ISA 201VG adjuvant produced strong whole-MAP as well as MAP2698c-specific IFN-γ responses in a high proportion of the vaccinated sheep. The formulation caused less severe injection site lesions in comparison to other formulations. The findings from this study suggest that the MAP2698c + 201VG should be evaluated in a challenge trial to determine the efficacy of this vaccine candidate.

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“…However, ELISA data also suggests that MptD, or at least the epitope recognized by the fMptD phage, is not exposed on the surface of L. salivarius. It has already been shown that MptD expression on the cell surface of a recombinant host can be achieved using fMptD for both M. smegmatis and M. bovis BCG (Stratmann et al, 2004; Heinzmann et al, 2008); which could be due to the presence of mycobacterium specific chaperones facilitating appropriate presentation of the MptD protein (Goldstone et al, 2008). However, mptD is naturally present on a six gene operon ( mptA-F ) transcribed as a single polycistronic mRNA molecule [60] and in both the aforementioned studies, the recombinant hosts were not transformed with the mptD gene in isolation.…”

Section: Discussionmentioning

confidence: 99%

“…However, mptD is naturally present on a six gene operon ( mptA-F ) transcribed as a single polycistronic mRNA molecule [60] and in both the aforementioned studies, the recombinant hosts were not transformed with the mptD gene in isolation. The M. smegmatis harbored a vector including mptC-F genes of the mpt operon (Stratmann et al, 2004) while the M. bovis BCG host contained the entire operon integrated into the chromosome (Heinzmann et al, 2008). Consequently, it is possible that for effective MptD cell surface display, some ancillary Mpt proteins may also be required.…”

Section: Discussionmentioning

confidence: 99%

Enhanced expression of codon optimized Mycobacterium avium subsp. paratuberculosis antigens in Lactobacillus salivarius

Johnston

Bannantine

Govender

et al. 2014

Front. Cell. Infect. Microbiol.

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It is well documented that open reading frames containing high GC content show poor expression in A+T rich hosts. Specifically, G+C-rich codon usage is a limiting factor in heterologous expression of Mycobacterium avium subsp. paratuberculosis (MAP) proteins using Lactobacillus salivarius. However, re-engineering opening reading frames through synonymous substitutions can offset codon bias and greatly enhance MAP protein production in this host. In this report, we demonstrate that codon-usage manipulation of MAP2121c can enhance the heterologous expression of the major membrane protein (MMP), analogous to the form in which it is produced natively by MAP bacilli. When heterologously over-expressed, antigenic determinants were preserved in synthetic MMP proteins as shown by monoclonal antibody mediated ELISA. Moreover, MMP is a membrane protein in MAP, which is also targeted to the cellular surface of recombinant L. salivarius at levels comparable to MAP. Additionally, we previously engineered MAP3733c (encoding MptD) and show herein that MptD displays the tendency to associate with the cytoplasmic membrane boundary under confocal microscopy and the intracellularly accumulated protein selectively adheres to the MptD-specific bacteriophage fMptD. This work demonstrates there is potential for L. salivarius as a viable antigen delivery vehicle for MAP, which may provide an effective mucosal vaccine against Johne's disease.

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Mycobacterium avium subsp. paratuberculosis-specific mpt operon expressed in M. bovis BCG as vaccine candidate

Cited by 11 publications

References 29 publications

Primary transcriptomes of Mycobacterium avium subsp. paratuberculosis reveal proprietary pathways in tissue and macrophages

Primary transcriptomes of Mycobacterium avium subsp. paratuberculosis reveal proprietary pathways in tissue and macrophages

Cellular and humoral immune responses in sheep vaccinated with candidate antigens MAP2698c and MAP3567 from Mycobacterium avium subspecies paratuberculosis

Enhanced expression of codon optimized Mycobacterium avium subsp. paratuberculosis antigens in Lactobacillus salivarius

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