Screening of Mycobacterium avium subsp. paratuberculosis mutants for attenuation in a bovine monocyte-derived macrophage model

Lamont, Elise A.; Talaat, Adel M.; Coussens, Paul M.; Bannantine, John P.; Gröhn, Yrjö T.; Katani, Robab; Li, Ling ling; Kapur, Vivek; Sreevatsan, Srinand

doi:10.3389/fcimb.2014.00087

Cited by 23 publications

(31 citation statements)

References 44 publications

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“…Each dot represents one individual response, the middle line represents the average in each group, and the error bars represent SD from the mean in each group. the concentration of NO − and bactericidal activity has been reported in rats and chickens but not in mice and cattle (Weiss et al, 2002;Sun et al, 2008;Guimarães et al, 2011;Zhao et al, 2013;Lamont et al, 2014;Akhtar et al, 2016;Qin et al, 2016). Lack of the ability to reliably measure macrophage function, including the production of NO − , seems to be the source of inconsistency.…”

Section: Discussionmentioning

confidence: 99%

The effect of host genetics on in vitro performance of bovine monocyte-derived macrophages

Emam

Tabatabaei

Sargolzaei

et al. 2019

Journal of Dairy Science

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The dynamic interaction between the host and pathogens, along with environmental factors, influences the regulation of mammalian immune responses. Therefore, comprehensive in vivo immune-phenotyping during an active response to a pathogen can be complex and prone to confounding effects. Evaluating critical fundamental aspects of the immune system at a cellular level is an alternative approach to reduce this complexity. Therefore, the objective of the current study was to examine an in vitro model for functional phenotyping of bovine monocyte-derived macrophages (MDM), cells which play a crucial role at all phases of inflammation, as well influence downstream immune responses. As indicators of MDM function, phagocytosis and nitric oxide (NO −) production were tested in MDM of 16 cows in response to 2 common bacterial pathogens of dairy cows, Escherichia coli and Staphylococcus aureus. Notable functional variations were observed among the individuals (coefficient of variation: 33% for phagocytosis and 70% in the production of NO −). The rank correlation analysis revealed a significant, positive, and strong correlation (rho = 0.92) between NO − production in response to E. coli and S. aureus, and a positive but moderate correlation (rho = 0.58) between phagocytosis of E. coli and S. aureus. To gain further insight into this trait, another 58 cows were evaluated solely for NO − response against E. coli. The pedigree of the tested animals was added to the statistical model and the heritability was estimated to be 0.776. Overall, the finding of this study showed a strong effect of host genetics on the in vitro activities of MDM and the possibility of ranking Holstein cows based on the in vitro functional variation of MDM.

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Section: Discussionmentioning

confidence: 99%

The effect of host genetics on in vitro performance of bovine monocyte-derived macrophages

Emam

Tabatabaei

Sargolzaei

et al. 2019

Journal of Dairy Science

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“…Strong evidence for the transposon insertions affecting the functions of Mod operon (e.g., MAP1566 or MAP1567) is provided by the similar properties of both mutants in MDMs, mice (Bannantine et al, 2014 ) and goats (Hines et al, 2014 ). These mutants were attenuated in C57BL/6 mice but protective against wild type challenge, displayed low skin reactivity in goats and showed significant attenuation in MDM assays as analyzed elsewhere (Lamont et al, 2014 ). However, in this current analysis, 30H9 and STM68 were comparable to the wild type strain.…”

Section: Discussionmentioning

confidence: 77%

“…Isolate 22F4 carries an insertion within the gene encoding the Lsr2 protein that has been implicated in various roles: dominant T cell responses in MTB (Stewart et al, 2002 ) and biofilm formation and colony morphology in Msmeg (Chen et al, 2006 ). Furthermore, a MAP lsr2 deletion mutant has been constructed (Park et al, 2008 ) that showed similar properties and a marginal degree of attenuation in MDMs to 22F4 (Lamont et al, 2014 ), thus providing strong evidence that the transposon insertion, that is internal to lsr2 , does disrupt the function of this gene.…”

Section: Discussionmentioning

confidence: 99%

“…However, in this current analysis, 30H9 and STM68 were comparable to the wild type strain. Interestingly, another MAP1566 Tn 5367 mutant (WAg906), derived from the MAP wild type strain 989 (Cavaignac et al, 2000 ), was shown to be attenuated based on the average slope values obtained in long-term MDM assays as well as BALB/c mice at 12 weeks post-infection (Scandurra et al, 2010 ; Bannantine et al, 2014 ; Lamont et al, 2014 ). However, in protection challenge experiments, 30H9 and STM68 showed good protection in C57BL/6 mice against wild type challenge, while WAg906 failed to protect BALB/c mice.…”

Section: Discussionmentioning

confidence: 99%

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Generation and screening of a comprehensive Mycobacterium avium subsp. paratuberculosis transposon mutant bank

Rathnaiah

Lamont

Harris

et al. 2014

Front. Cell. Infect. Microbiol.

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Mycobacterium avium subsp. paratuberculosis (MAP) is the etiologic agent of Johne's Disease in ruminants. This enteritis has significant economic impact and worldwide distribution. Vaccination is one of the most cost effective infectious disease control measures. Unfortunately, current vaccines reduce clinical disease and shedding, but are of limited efficacy and do not provide long-term protective immunity. Several strategies have been followed to mine the MAP genome for virulence determinants that could be applied to vaccine and diagnostic assay development. In this study, a comprehensive mutant bank of 13,536 MAP K-10 Tn5367 mutants (P > 95%) was constructed and screened in vitro for phenotypes related to virulence. This strategy was designated to maximize identification of genes important to MAP pathogenesis without relying on studies of other mycobacterial species that may not translate into similar effects in MAP. This bank was screened for mutants with colony morphology alterations, susceptibility to D-cycloserine, impairment in siderophore production or secretion, reduced cell association, and decreased biofilm and clump formation. Mutants with interesting phenotypes were analyzed by PCR, Southern blotting and DNA sequencing to determine transposon insertion sites. These insertion sites mapped upstream from the MAP1152-MAP1156 cluster, internal to either the Mod operon gene MAP1566 or within the coding sequence of lsr2, and several intergenic regions. Growth curves in broth cultures, invasion assays and kinetics of survival and replication in primary bovine macrophages were also determined. The ability of vectors carrying Tn5370 to generate stable MAP mutants was also investigated.

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“…Scandurra et al (2010) were able to classify the strains according to their virulence and made a preliminary selection of vaccine candidates in a well-defined bovine monocytederived macrophage model. However, some studies have highlighted the limitations of the macrophage model in the initial screening of Map virulence and in predicting vaccine candidates (Bannantine et al, 2014b;Lamont et al, 2014). Vaccine trials in goat, sheep, deer or bovine hosts are highly costly and thus the use of small animal models such as mice facilitate comparisons between different vaccine trials.…”

Section: Introductionmentioning

confidence: 99%

Protection efficacy of Argentinian isolates of Mycobacterium avium subsp. paratuberculosis with different genotypes and virulence in a murine model

Olivieri

Moyano

Travería³

et al. 2018

Research in Veterinary Science

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Protection efficacy of Argentinian isolates of Mycobacterium avium subsp. paratuberculosis with different genotypes and virulence in a murine model. AbstractParatuberculosis is a chronic disease caused by Mycobacterium avium subsp.paratuberculosis (Map). The disease causes economic losses and, therefore, it is imperative to follow proper control strategies, which should include an effective vaccine. Several strategies have assessed the virulence and immune response of Map strains that could be used as a vaccine. This study evaluates the degree of virulence, immune response, and protection of Argentinian strains of Map with different genotype in a murine model. Four local isolates (Cattle type) with different genotypes (analyzed by MIRU-VNTR and SSRs) were selected and evaluated in a virulence assay in BALB/c mice. This assay allowed us to differentiate virulent and low-virulence Map strains. The less virulent strains (1543/481 and A162) failed to induce a significant production of the proinflammatory cytokine IFNg, whereas the virulent strain 6611 established infection along with a proinflammatory immune response. On the other hand, the virulent strain 1347/498 was efficient in establishing a persistent infection, but failed to promote an important Th1 response compared with 6611 at the evaluated time.We selected the low-virulence strain 1543/498 as a live vaccine and the virulent strain 6611 as a live and inactivated vaccine in a protection assay in mice. Strain 1543/481 failed to protect the animals from challenge, whereas strain 6611, in its live and inactivated form, significantly reduced the CFUs count in the infected mice, although ACCEPTED MANUSCRIPT

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Screening of Mycobacterium avium subsp. paratuberculosis mutants for attenuation in a bovine monocyte-derived macrophage model

Cited by 23 publications

References 44 publications

The effect of host genetics on in vitro performance of bovine monocyte-derived macrophages

The effect of host genetics on in vitro performance of bovine monocyte-derived macrophages

Generation and screening of a comprehensive Mycobacterium avium subsp. paratuberculosis transposon mutant bank

Protection efficacy of Argentinian isolates of Mycobacterium avium subsp. paratuberculosis with different genotypes and virulence in a murine model

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