title: 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.
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.
authors.affiliation: Department of Animal Health, Basque Institute for Agricultural Research and Development, NEIKER-Tecnalia, Technological Park of Bizkaia, Bizkaia, Spain.
authors.forename: Naiara
authors.initials: N
authors.lastname: Abendaño
authors.affiliation: Department of Animal Health, Basque Institute for Agricultural Research and Development, NEIKER-Tecnalia, Technological Park of Bizkaia, Bizkaia, Spain.
authors.forename: Lyudmila
authors.initials: L
authors.lastname: Tyukalova
authors.affiliation: Department of Animal Health, Basque Institute for Agricultural Research and Development, NEIKER-Tecnalia, Technological Park of Bizkaia, Bizkaia, Spain.
authors.forename: Jesse F
authors.initials: JF
authors.lastname: Barandika
authors.affiliation: Department of Animal Health, Department of Agriculture of the Regional Government of the Principality of Asturias, SERIDA, Deva, Asturias, Spain.
authors.forename: Ana
authors.initials: A
authors.lastname: Balseiro
authors.affiliation: Department of Animal Health, Basque Institute for Agricultural Research and Development, NEIKER-Tecnalia, Technological Park of Bizkaia, Bizkaia, Spain.
authors.forename: Iker A
authors.initials: IA
authors.lastname: Sevilla
authors.affiliation: Department of Animal Health, Basque Institute for Agricultural Research and Development, NEIKER-Tecnalia, Technological Park of Bizkaia, Bizkaia, Spain.
authors.forename: Joseba M
authors.initials: JM
authors.lastname: Garrido
authors.affiliation: Department of Animal Health, Basque Institute for Agricultural Research and Development, NEIKER-Tecnalia, Technological Park of Bizkaia, Bizkaia, Spain.
authors.forename: Ramon A
authors.initials: RA
authors.lastname: Juste
authors.affiliation: Department of Animal Health, Basque Institute for Agricultural Research and Development, NEIKER-Tecnalia, Technological Park of Bizkaia, Bizkaia, Spain.
authors.forename: Marta
authors.initials: M
authors.lastname: Alonso-Hearn
mesh_terms: D000818:Animals; D017209:Apoptosis; D002417:Cattle; D005786:Gene Expression Regulation; D005838:Genotype; D006099:Granuloma / Q000276:immunology / Q000382:microbiology*; D058507:Host Specificity; D007249:Inflammation / Q000378:metabolism / Q000382:microbiology; D008264:Macrophages / Q000166:cytology* / Q000276:immunology / Q000378:metabolism; D050296:Microbial Viability; D009000:Monocytes / Q000166:cytology; D016927:Mycobacterium avium subsp. paratuberculosis / Q000302:isolation & purification* / Q000502:physiology*; D010641:Phenotype; D012756:Sheep / Q000382:microbiology
references.citation: Biomed Res Int. 2013;2013:908348
references.pmid: 23509800
references.citation: J Clin Microbiol. 2012 Aug;50(8):2609-17
references.pmid: 22649014
references.citation: Prev Vet Med. 1999 Jun 11;40(3-4):179-92
references.pmid: 10423773
references.citation: Vet Immunol Immunopathol. 2001 Oct;82(3-4):139-51
references.pmid: 11587730
references.citation: Vet Immunol Immunopathol. 2006 Mar 15;110(1-2):109-20
references.pmid: 16256206
references.citation: Nat Rev Immunol. 2012 Apr 20;12(5):352-66
references.pmid: 22517424
references.citation: J Infect Dis. 2005 Feb 1;191(3):453-60
references.pmid: 15633105
references.citation: J Pathol. 2007 Jan;211(1):76-85
references.pmid: 17115379
references.citation: PLoS One. 2013;8(1):e53657
references.pmid: 23308269
references.citation: Aust Vet J. 2001 Jul;79(7):484-91
references.pmid: 11549048
references.citation: Vet Q. 2000 Oct;22(4):200-4
references.pmid: 11087130
references.citation: Prev Vet Med. 2009 Jan 1;88(1):1-14
references.pmid: 18817995
references.citation: Vet Immunol Immunopathol. 2009 Mar 15;128(1-3):44-52
references.pmid: 19022505
references.citation: Vet Microbiol. 2013 May 3;163(3-4):325-34
references.pmid: 23415474
references.citation: Infect Immun. 2003 Nov;71(11):6487-98
references.pmid: 14573671
references.citation: Vet Microbiol. 1995 Jul;45(2-3):211-7
references.pmid: 7571372
references.citation: Rev Sci Tech. 2005 Dec;24(3):1061-6
references.pmid: 16642774
references.citation: Vet Pathol. 2004 Jul;41(4):326-37
references.pmid: 15232132
references.citation: Infect Immun. 2010 Aug;78(8):3570-7
references.pmid: 20498259
references.citation: Cells Tissues Organs. 2003;174(3):110-6
references.pmid: 12835574
references.citation: BMC Microbiol. 2006 Feb 14;6:10
references.pmid: 16478544
references.citation: J Clin Microbiol. 2002 May;40(5):1798-804
references.pmid: 11980962
references.citation: Zoonoses Public Health. 2007;54(6-7):243-52
references.pmid: 17803513
references.citation: J Clin Microbiol. 1990 Jul;28(7):1591-6
references.pmid: 2166089
references.citation: Clin Microbiol Rev. 2001 Jul;14(3):489-512
references.pmid: 11432810
references.citation: J Vet Diagn Invest. 2005 Jul;17(4):354-9
references.pmid: 16130994
references.citation: J Pathol. 1997 Jan;181(1):25-30
references.pmid: 9071999
references.citation: Immunol Cell Biol. 2007 Feb-Mar;85(2):160-8
references.pmid: 17199112
references.citation: Immunol Cell Biol. 2000 Aug;78(4):334-41
references.pmid: 10947857
references.citation: Cell Microbiol. 2004 May;6(5):423-33
references.pmid: 15056213
references.citation: Mol Cell Probes. 2001 Jun;15(3):139-45
references.pmid: 11352594
references.citation: J Immunol. 2007 Mar 1;178(5):3161-9
references.pmid: 17312164
references.citation: Comp Immunol Microbiol Infect Dis. 2003 Jul;26(4):269-83
references.pmid: 12676127
references.citation: Nat Immunol. 2009 Sep;10(9):943-8
references.pmid: 19692995
references.citation: Infect Immun. 2002 Oct;70(10):5556-61
references.pmid: 12228282
references.citation: Vet Res. 2012 Mar 28;43:25
references.pmid: 22455317
references.citation: Appl Environ Microbiol. 2008 Jan;74(1):275-85
references.pmid: 17981953
references.citation: BMC Microbiol. 2007 Mar 12;7:18
references.pmid: 17352818
references.citation: Physiol Genomics. 2006 Dec 13;28(1):67-75
references.pmid: 17062651
references.citation: Vet Res. 2014 Jan 16;45:5
references.pmid: 24428881
references.citation: Brief Funct Genomics. 2010 May;9(3):220-37
references.pmid: 20495212