title: Granzyme B-expressing neutrophils correlate with bacterial load in granulomas from Mycobacterium tuberculosis-infected cynomolgus macaques.
abstract: The role of neutrophils in tuberculosis (TB), and whether neutrophils express granzyme B (grzB), a pro-apoptotic enzyme associated with cytotoxic T cells, is controversial. We examined neutrophils in peripheral blood (PB) and lung granulomas of Mycobacterium tuberculosis-infected cynomolgus macaques and humans to determine whether mycobacterial products or pro-inflammatory factors induce neutrophil grzB expression. We found large numbers of grzB-expressing neutrophils in macaque and human granulomas and these cells contained more grzB+ granules than T cells. Higher neutrophil, but not T cell, grzB expression correlated with increased bacterial load. Although unstimulated PB neutrophils lacked grzB expression, grzB expression increased upon exposure to M.tuberculosis bacilli, M.tuberculosis culture filtrate protein or lipopolysaccharide from Escherichia coli. Perforin is required for granzyme-mediated cytotoxicity by T cells, but was not observed in PB or granuloma neutrophils. Nonetheless, stimulated PB neutrophils secreted grzB as determined by enzyme-linked immunospot assays. Purified grzB was not bactericidal or bacteriostatic, suggesting secreted neutrophil grzB acts on extracellular targets, potentially enhancing neutrophil migration through extracellular matrix and regulating apoptosis or activation in other cell types. These data indicate mycobacterial products and the pro-inflammatory environment of granulomas up-regulates neutrophil grzB expression and suggests a previously unappreciated aspect of neutrophil biology in TB.
authors: Department of Microbiology and Molecular Genetics, University of Pittsburgh, Pittsburgh, PA, USA.
authors: Joshua T
authors: JT
authors: Mattila
authors: Department of Microbiology and Molecular Genetics, University of Pittsburgh, Pittsburgh, PA, USA.
authors: Pauline
authors: P
authors: Maiello
authors: Department of Immunology, University of Pittsburgh, Pittsburgh, PA, USA.
authors: Tao
authors: T
authors: Sun
authors: Tuberculosis Research Section, Laboratory of Clinical Infectious Diseases, NIAID, NIH, Bethesda, MD, USA.
authors: Laura E
authors: LE
authors: Via
authors: Department of Microbiology and Molecular Genetics, University of Pittsburgh, Pittsburgh, PA, USA.|Department of Immunology, University of Pittsburgh, Pittsburgh, PA, USA.
authors: JoAnne L
authors: JL
authors: Flynn
mesh_terms: D000818:Animals; D058491:Bacterial Load; D004195:Disease Models, Animal; D058501:Enzyme-Linked Immunospot Assay; D005434:Flow Cytometry; D020869:Gene Expression Profiling; D006099:Granuloma / Q000382:microbiology / Q000473:pathology*; D053804:Granzymes / Q000032:analysis* / Q000378:metabolism; D006801:Humans; D008252:Macaca fascicularis; D009169:Mycobacterium tuberculosis / Q000302:isolation & purification*; D009504:Neutrophils / Q000737:chemistry*; D014376:Tuberculosis / Q000382:microbiology / Q000473:pathology*
chemical_list: D053804:Granzymes
references: Genome Biol. 2006;7(10):R100
references: 17076895
references: Cell Microbiol. 2007 Feb;9(2):382-96
references: 16925784
references: Am J Respir Crit Care Med. 2007 Mar 1;175(5):464-72
references: 17138956
references: Toxicol Pathol. 2007 Jan;35(1):111-5
references: 17325979
references: J Biol Chem. 2007 Mar 16;282(11):8157-64
references: 17224449
references: Immunol Lett. 2007 Jul 31;111(1):57-65
references: 17604848
references: Inflammation. 2007 Oct;30(5):178-88
references: 17624583
references: Circ Res. 2008 Jan 4;102(1):32-41
references: 17975113
references: Eur J Immunol. 2008 Feb;38(2):437-47
references: 18203135
references: Mol Immunol. 2008 Mar;45(6):1761-6
references: 17996944
references: J Infect. 2008 May;56(5):340-7
references: 18359089
references: Int J Biochem Cell Biol. 2008;40(6-7):1317-33
references: 18180196
references: Int J Chron Obstruct Pulmon Dis. 2008;3(2):253-68
references: 18686734
references: J Biol Chem. 2008 Aug 15;283(33):22498-504
references: 18577516
references: J Immunol. 2008 Dec 15;181(12):8595-603
references: 19050279
references: J Immunol. 2009 Aug 15;183(4):2602-9
references: 19620298
references: Infect Immun. 1997 Apr;65(4):1317-20
references: 9119468
references: Eur Respir J. 1998 Jun;11(6):1227-31
references: 9657559
references: Cell Signal. 2005 Mar;17(3):385-94
references: 15567069
references: Arterioscler Thromb Vasc Biol. 2004 Dec;24(12):2245-50
references: 15472125
references: Immunol Res. 2005;32(1-3):211-5
references: 16106072
references: Blood. 2005 Sep 1;106(5):1843-50
references: 15886329
references: Blood. 2005 Oct 15;106(8):2871-8
references: 15998831
references: Blood. 2005 Nov 15;106(10):3474-82
references: 16037389
references: J Biol Chem. 2006 May 12;281(19):13525-32
references: 16524880
references: Nat Rev Immunol. 2006 Jul;6(7):541-50
references: 16799473
references: J Biol Chem. 2006 Jul 14;281(28):19781-92
references: 16690621
references: Cell Death Differ. 2010 Apr;17(4):596-606
references: 20139894
references: Immunol Rev. 2010 May;235(1):105-16
references: 20536558
references: J Dermatol Sci. 2010 Aug;59(2):129-35
references: 20621450
references: Nature. 2010 Aug 19;466(7309):973-7
references: 20725040
references: J Clin Invest. 2010 Sep;120(9):3365-76
references: 20679732
references: Int J Oncol. 2010 Dec;37(6):1361-78
references: 21042704
references: J Immunol. 2011 Jun 15;186(12):7110-9
references: 21555529
references: Mol Cell. 2011 Oct 21;44(2):265-78
references: 22017873
references: J Exp Med. 2011 Oct 24;208(11):2251-62
references: 21967766
references: Trends Immunol. 2012 Jan;33(1):14-25
references: 22094048
references: Cell Host Microbe. 2012 Jan 19;11(1):81-90
references: 22264515
references: PLoS One. 2012;7(2):e30626
references: 22359547
references: J Immunol. 2012 May 1;188(9):4476-87
references: 22461690
references: Front Biosci (Landmark Ed). 2012;17:2278-83
references: 22652778
references: Proc Natl Acad Sci U S A. 2012 Aug 28;109(35):14188-93
references: 22826237
references: Cell Host Microbe. 2012 Sep 13;12(3):301-12
references: 22980327
references: Trends Mol Med. 2012 Dec;18(12):732-41
references: 23099058
references: PLoS One. 2012;7(12):e50345
references: 23251364
references: Antioxid Redox Signal. 2013 Feb 20;18(6):642-60
references: 22881869
references: Am J Respir Cell Mol Biol. 2013 May;48(5):531-9
references: 23328639
references: J Immunol. 2013 Jul 15;191(2):773-84
references: 23749634
references: J Immunol. 2013 Oct 1;191(7):3818-26
references: 23997221
references: Tuberculosis (Edinb). 2013 Nov;93(6):675-87
references: 23988278
references: Nat Med. 2014 Jan;20(1):75-9
references: 24336248
references: Ther Adv Respir Dis. 2009 Jun;3(3):113-29
references: 19638369
references: Infect Immun. 2009 Oct;77(10):4631-42
references: 19620341
references: Lab Invest. 2009 Nov;89(11):1195-220
references: 19770840
references: Chest. 2010 Jan;137(1):122-8
references: 19749004
references: Am J Pathol. 2010 Feb;176(2):1038-49
references: 20035050
references: Arthritis Rheum. 2010 Feb;62(2):340-50
references: 20112395
references: Cell Death Differ. 2010 Apr;17(4):616-23
references: 20075940
references: Annu Rev Pathol. 2014;9:181-218
references: 24050624
references: Immunol Lett. 2014 Jul;160(1):23-32
references: 24694750
references: Proc Natl Acad Sci U S A. 2000 Apr 11;97(8):4204-8
references: 10760288
references: J Immunol. 2000 Jul 1;165(1):353-63
references: 10861072
references: Proc Natl Acad Sci U S A. 2000 Aug 1;97(16):8841-8
references: 10922044
references: Chest. 2002 Feb;121(2):519-26
references: 11834667
references: Nat Rev Immunol. 2002 Jun;2(6):401-9
references: 12093006
references: Nat Rev Immunol. 2002 Oct;2(10):735-47
references: 12360212
references: Mod Pathol. 2003 May;16(5):460-70
references: 12748253
references: J Rheumatol. 2003 Aug;30(8):1799-810
references: 12913938
references: Blood. 2004 Feb 1;103(3):1099-104
references: 14512315
references: Infect Immun. 2004 Mar;72(3):1804-6
references: 14977991
references: Blood. 2004 Aug 1;104(3):905-6; author reply 907-8
references: 15265799
references: Blood. 2004 Aug 1;104(3):906-7; author reply 907-8
references: 15265800
references: J Immunol. 2004 Aug 15;173(4):2746-54
references: 15294993
references: Chest. 1992 Jul;102(1):54-9
references: 1623796
references: Proc Natl Acad Sci U S A. 1992 Dec 15;89(24):12013-7
references: 1465432
references: Am J Respir Crit Care Med. 1996 Feb;153(2):799-804
references: 8564135
references: Clin Exp Immunol. 2006 Nov;146(2):243-52
references: 17034576