Limits...
Mycobacterium Avium subsp. paratuberculosis isolates induce in vitro granuloma formation and show successful survival phenotype, common anti-inflammatory and antiapoptotic responses within ovine macrophages regardless of genotype or host of origin.

Abendaño N, Tyukalova L, Barandika JF, Balseiro A, Sevilla IA, Garrido JM, Juste RA, Alonso-Hearn M - PLoS ONE (2014)

Bottom Line: After 7 d of infection, variations in the estimated log10 CFUs between all the tested isolates were not statistically significant.In addition, ovine MDMs exhibited enhanced anti-inflammatory, antiapoptotic and antidestructive responses when infected with two ovine isolates of distinct genotype (C and S) or with two C-type isolates from distinct hosts (cattle and sheep); which correlated with the successful survival of these isolates within ovine MDMs. A second objective was to study, based on an in vitro granuloma model, latter stages of the infection by investigating the capacity of two Map isolates from cattle and sheep to trigger formation of microgranulomas.In summary, our results demonstrated that Map isolates from cattle, sheep, goats, deer, fallow-deer and wild boar were able not only to initiate but also to establish a successful infection in ovine macrophages regardless of genotype.

View Article: PubMed Central - PubMed

Affiliation: Department of Animal Health, Basque Institute for Agricultural Research and Development, NEIKER-Tecnalia, Technological Park of Bizkaia, Bizkaia, Spain.

ABSTRACT
The analysis of the early macrophage responses, including bacterial growth within macrophages, represents a powerful tool to characterize the virulence of clinical isolates of Mycobcaterium avium susbp. paratuberculosis (Map). The present study represents the first assessment of the intracellular behaviour in ovine monocyte-derived macrophages (MDMs) of Map isolates representing distinct genotypes (C, S and B), and isolated from cattle, sheep, goat, fallow deer, deer, and wild boar. Intracellular growth and survival of the selected isolates in ovine MDMs was assessed by quantification of CFUs inside of the host cells at 2 h p.i. (day 0) and 7 d p. i. using an automatic liquid culture system (Bactec MGIT 960). Variations in bacterial counts over 7 days from the baseline were small, in a range between 1.63 to 1.05-fold. After 7 d of infection, variations in the estimated log10 CFUs between all the tested isolates were not statistically significant. In addition, ovine MDMs exhibited enhanced anti-inflammatory, antiapoptotic and antidestructive responses when infected with two ovine isolates of distinct genotype (C and S) or with two C-type isolates from distinct hosts (cattle and sheep); which correlated with the successful survival of these isolates within ovine MDMs. A second objective was to study, based on an in vitro granuloma model, latter stages of the infection by investigating the capacity of two Map isolates from cattle and sheep to trigger formation of microgranulomas. Upon 10 d p.i., both Map isolates were able to induce the formation of granulomas comparable to the granulomas observed in clinical specimens with respect to the cellular components involved. In summary, our results demonstrated that Map isolates from cattle, sheep, goats, deer, fallow-deer and wild boar were able not only to initiate but also to establish a successful infection in ovine macrophages regardless of genotype.

Show MeSH

Related in: MedlinePlus

Number of microgranulomes generated in vitro 5 and 10 days after infection of ovine PBMCs with (A) the K10 reference strain and with (B) the ovine 2349/06-1 isolate.Aggregate numbers were estimated under a light microscope. The mean aggregate number estimated by triplicate is shown for each MOI (1:8, 1:16 and 1:33) ± SD.
© Copyright Policy
Related In: Results  -  Collection

License
getmorefigures.php?uid=PMC4128652&req=5

pone-0104238-g003: Number of microgranulomes generated in vitro 5 and 10 days after infection of ovine PBMCs with (A) the K10 reference strain and with (B) the ovine 2349/06-1 isolate.Aggregate numbers were estimated under a light microscope. The mean aggregate number estimated by triplicate is shown for each MOI (1:8, 1:16 and 1:33) ± SD.

Mentions: The number of granuloma-like aggregates formed in response to the infection with both Map isolates at 5 and 10 days post-infection is shown in Figure 3. Statistical analysis of the data showed significant variations between the number of aggregates generated after 5 days of infection with the K10 reference strain and the 2349/06 isolate at the highest MOI (1∶8), with the K10 strain inducing the formation of more aggregates than the ovine isolate (P<0.01). However, after 10 days of infection the number of aggregates formed by both strains was not significantly different at any of the 3 assessed MOIs. The number of aggregates generated by both strains at the highest and lowest MOI (1∶8 and 1∶33) was significantly different at 10 days p. i. Significant differences in the number of aggregates triggered by both strains between days 5 and 10 were not observed which suggested that most of the granulomas were formed during the first five days of the infection with both isolates. Uninfected cells showed no granuloma formation.


Mycobacterium Avium subsp. paratuberculosis isolates induce in vitro granuloma formation and show successful survival phenotype, common anti-inflammatory and antiapoptotic responses within ovine macrophages regardless of genotype or host of origin.

Abendaño N, Tyukalova L, Barandika JF, Balseiro A, Sevilla IA, Garrido JM, Juste RA, Alonso-Hearn M - PLoS ONE (2014)

Number of microgranulomes generated in vitro 5 and 10 days after infection of ovine PBMCs with (A) the K10 reference strain and with (B) the ovine 2349/06-1 isolate.Aggregate numbers were estimated under a light microscope. The mean aggregate number estimated by triplicate is shown for each MOI (1:8, 1:16 and 1:33) ± SD.
© Copyright Policy
Related In: Results  -  Collection

License
Show All Figures
getmorefigures.php?uid=PMC4128652&req=5

pone-0104238-g003: Number of microgranulomes generated in vitro 5 and 10 days after infection of ovine PBMCs with (A) the K10 reference strain and with (B) the ovine 2349/06-1 isolate.Aggregate numbers were estimated under a light microscope. The mean aggregate number estimated by triplicate is shown for each MOI (1:8, 1:16 and 1:33) ± SD.
Mentions: The number of granuloma-like aggregates formed in response to the infection with both Map isolates at 5 and 10 days post-infection is shown in Figure 3. Statistical analysis of the data showed significant variations between the number of aggregates generated after 5 days of infection with the K10 reference strain and the 2349/06 isolate at the highest MOI (1∶8), with the K10 strain inducing the formation of more aggregates than the ovine isolate (P<0.01). However, after 10 days of infection the number of aggregates formed by both strains was not significantly different at any of the 3 assessed MOIs. The number of aggregates generated by both strains at the highest and lowest MOI (1∶8 and 1∶33) was significantly different at 10 days p. i. Significant differences in the number of aggregates triggered by both strains between days 5 and 10 were not observed which suggested that most of the granulomas were formed during the first five days of the infection with both isolates. Uninfected cells showed no granuloma formation.

Bottom Line: After 7 d of infection, variations in the estimated log10 CFUs between all the tested isolates were not statistically significant.In addition, ovine MDMs exhibited enhanced anti-inflammatory, antiapoptotic and antidestructive responses when infected with two ovine isolates of distinct genotype (C and S) or with two C-type isolates from distinct hosts (cattle and sheep); which correlated with the successful survival of these isolates within ovine MDMs. A second objective was to study, based on an in vitro granuloma model, latter stages of the infection by investigating the capacity of two Map isolates from cattle and sheep to trigger formation of microgranulomas.In summary, our results demonstrated that Map isolates from cattle, sheep, goats, deer, fallow-deer and wild boar were able not only to initiate but also to establish a successful infection in ovine macrophages regardless of genotype.

View Article: PubMed Central - PubMed

Affiliation: Department of Animal Health, Basque Institute for Agricultural Research and Development, NEIKER-Tecnalia, Technological Park of Bizkaia, Bizkaia, Spain.

ABSTRACT
The analysis of the early macrophage responses, including bacterial growth within macrophages, represents a powerful tool to characterize the virulence of clinical isolates of Mycobcaterium avium susbp. paratuberculosis (Map). The present study represents the first assessment of the intracellular behaviour in ovine monocyte-derived macrophages (MDMs) of Map isolates representing distinct genotypes (C, S and B), and isolated from cattle, sheep, goat, fallow deer, deer, and wild boar. Intracellular growth and survival of the selected isolates in ovine MDMs was assessed by quantification of CFUs inside of the host cells at 2 h p.i. (day 0) and 7 d p. i. using an automatic liquid culture system (Bactec MGIT 960). Variations in bacterial counts over 7 days from the baseline were small, in a range between 1.63 to 1.05-fold. After 7 d of infection, variations in the estimated log10 CFUs between all the tested isolates were not statistically significant. In addition, ovine MDMs exhibited enhanced anti-inflammatory, antiapoptotic and antidestructive responses when infected with two ovine isolates of distinct genotype (C and S) or with two C-type isolates from distinct hosts (cattle and sheep); which correlated with the successful survival of these isolates within ovine MDMs. A second objective was to study, based on an in vitro granuloma model, latter stages of the infection by investigating the capacity of two Map isolates from cattle and sheep to trigger formation of microgranulomas. Upon 10 d p.i., both Map isolates were able to induce the formation of granulomas comparable to the granulomas observed in clinical specimens with respect to the cellular components involved. In summary, our results demonstrated that Map isolates from cattle, sheep, goats, deer, fallow-deer and wild boar were able not only to initiate but also to establish a successful infection in ovine macrophages regardless of genotype.

Show MeSH
Related in: MedlinePlus