title: Cryptococcus neoformans-induced macrophage lysosome damage crucially contributes to fungal virulence.
abstract: Upon ingestion by macrophages, Cryptococcus neoformans can survive and replicate intracellularly unless the macrophages become classically activated. The mechanism enabling intracellular replication is not fully understood; neither are the mechanisms that allow classical activation to counteract replication. C. neoformans-induced lysosome damage was observed in infected murine bone marrow-derived macrophages, increased with time, and required yeast viability. To demonstrate lysosome damage in the infected host, we developed a novel flow cytometric method for measuring lysosome damage. Increased lysosome damage was found in C. neoformans-containing lung cells compared with C. neoformans-free cells. Among C. neoformans-containing myeloid cells, recently recruited cells displayed lower damage than resident cells, consistent with the protective role of recruited macrophages. The magnitude of lysosome damage correlated with increased C. neoformans replication. Experimental induction of lysosome damage increased C. neoformans replication. Activation of macrophages with IFN-γ abolished macrophage lysosome damage and enabled increased killing of C. neoformans. We conclude that induction of lysosome damage is an important C. neoformans survival strategy and that classical activation of host macrophages counters replication by preventing damage. Thus, therapeutic strategies that decrease lysosomal damage, or increase resistance to such damage, could be valuable in treating cryptococcal infections.
authors: Division of Pulmonary and Critical Care Medicine, University of Michigan Health System, Ann Arbor, MI 48109;
authors: Michael J
authors: MJ
authors: Davis
authors: Division of Pulmonary and Critical Care Medicine, University of Michigan Health System, Ann Arbor, MI 48109; Graduate Program in Immunology, University of Michigan, Ann Arbor, MI 48109;
authors: Alison J
authors: AJ
authors: Eastman
authors: Division of Pulmonary and Critical Care Medicine, University of Michigan Health System, Ann Arbor, MI 48109; VA Ann Arbor Healthcare System, Ann Arbor, MI 48105;
authors: Yafeng
authors: Y
authors: Qiu
authors: Department of Microbiology and Immunology, University of Michigan Medical School, Ann Arbor, MI 48109; and.
authors: Brian
authors: B
authors: Gregorka
authors: Department of Microbiology and Immunology, University of Nevada School of Medicine, Reno, NV 89557.
authors: Thomas R
authors: TR
authors: Kozel
authors: Division of Pulmonary and Critical Care Medicine, University of Michigan Health System, Ann Arbor, MI 48109; VA Ann Arbor Healthcare System, Ann Arbor, MI 48105;
authors: John J
authors: JJ
authors: Osterholzer
authors: Division of Pulmonary and Critical Care Medicine, University of Michigan Health System, Ann Arbor, MI 48109; Graduate Program in Immunology, University of Michigan, Ann Arbor, MI 48109; VA Ann Arbor Healthcare System, Ann Arbor, MI 48105;
authors: Jeffrey L
authors: JL
authors: Curtis
authors: Department of Microbiology and Immunology, University of Michigan Medical School, Ann Arbor, MI 48109; and.
authors: Joel A
authors: JA
authors: Swanson
authors: Division of Pulmonary and Critical Care Medicine, University of Michigan Health System, Ann Arbor, MI 48109; Graduate Program in Immunology, University of Michigan, Ann Arbor, MI 48109; VA Ann Arbor Healthcare System, Ann Arbor, MI 48105; olszewsm@med.umich.edu.
authors: Michal A
authors: MA
authors: Olszewski
mesh_terms: D000818:Animals; D003453:Cryptococcosis / Q000188:drug therapy* / Q000276:immunology / Q000382:microbiology / Q000473:pathology; D003455:Cryptococcus neoformans / Q000276:immunology / Q000472:pathogenicity*; D054884:Host-Pathogen Interactions; D007371:Interferon-gamma / Q000494:pharmacology*; D008027:Light; D008168:Lung / Q000276:immunology / Q000382:microbiology / Q000473:pathology; D008172:Lung Diseases, Fungal / Q000188:drug therapy* / Q000276:immunology / Q000382:microbiology / Q000473:pathology; D008247:Lysosomes / Q000187:drug effects* / Q000382:microbiology / Q000473:pathology / Q000528:radiation effects; D008262:Macrophage Activation / Q000187:drug effects; D008264:Macrophages / Q000187:drug effects* / Q000276:immunology / Q000382:microbiology / Q000473:pathology; D051379:Mice; D008807:Mice, Inbred BALB C; D055668:Photochemical Processes; D061251:Primary Cell Culture; D014774:Virulence
chemical_list: D007371:Interferon-gamma
references: Nat Immunol. 2008 Aug;9(8):847-56
references: 18604214
references: Infect Immun. 2008 Oct;76(10):4764-71
references: 18678670
references: Nat Rev Mol Cell Biol. 2008 Oct;9(10):781-95
references: 18813294
references: Infect Immun. 2009 Aug;77(8):3450-7
references: 19487474
references: Infect Immun. 2009 Sep;77(9):3749-58
references: 19564388
references: Eukaryot Cell. 2009 Sep;8(9):1373-80
references: 19465562
references: Am J Pathol. 2009 Dec;175(6):2489-500
references: 19893050
references: J Immunol. 1988 May 1;140(9):3006-13
references: 2834450
references: N Engl J Med. 1989 Sep 21;321(12):794-9
references: 2671735
references: J Cell Sci. 1989 Sep;94 ( Pt 1):135-42
references: 2613767
references: J Exp Med. 1991 Apr 1;173(4):793-800
references: 1672543
references: J Leukoc Biol. 1994 Jan;55(1):35-42
references: 7904293
references: Eur J Clin Microbiol Infect Dis. 2008 Oct;27(10):937-43
references: 18449582
references: J Immunol. 2008 Nov 1;181(9):6349-60
references: 18941226
references: Oncogene. 2008 Oct 27;27(50):6434-51
references: 18955971
references: Am J Pathol. 2009 Mar;174(3):932-43
references: 19218345
references: AIDS. 2009 Feb 20;23(4):525-30
references: 19182676
references: Cell Host Microbe. 2009 May 8;5(5):487-97
references: 19454352
references: Nature. 2009 May 21;459(7245):433-6
references: 19339971
references: Am J Pathol. 2010 Feb;176(2):774-85
references: 20056835
references: Nature. 2010 Jan 28;463(7280):549-53
references: 20111001
references: PLoS One. 2010;5(4):e10008
references: 20368800
references: PLoS Pathog. 2010;6(8):e1001041
references: 20714349
references: Future Microbiol. 2010 Aug;5(8):1269-88
references: 20722603
references: J Leukoc Biol. 2010 Oct;88(4):813-22
references: 20587739
references: Am J Pathol. 2011 Jan;178(1):198-211
references: 21224057
references: Infect Immun. 2011 Mar;79(3):1098-106
references: 21149584
references: MBio. 2011;2(4). pii: e00167-11. doi: 10.1128/mBio.00167-11
references: 21828219
references: MBio. 2011;2(4). pii: e00158-11. doi: 10.1128/mBio.00158-11
references: 21828220
references: EMBO J. 2011 Aug 31;30(17):3481-500
references: 21878991
references: J Immunol. 2012 Jan 1;188(1):334-44
references: 22102723
references: J Immunol. 2012 Jul 1;189(1):112-9
references: 22615206
references: Sci Rep. 2012;2:739
references: 23074646
references: J Immunol. 2012 Nov 1;189(9):4488-95
references: 23002437
references: PLoS One. 2012;7(10):e47853
references: 23110112
references: ACS Nano. 2012 Dec 21;6(12):10792-9
references: 23146149
references: MBio. 2013;4(3):e00264-13
references: 23781069
references: Infect Immun. 2014 May;82(5):2059-67
references: 24595144
references: Infect Immun. 2000 Jul;68(7):4225-37
references: 10858240
references: Infect Immun. 2000 Nov;68(11):6147-53
references: 11035718
references: J Infect Dis. 2001 Mar 15;183(6):935-42
references: 11237811
references: Cancer Res. 2001 Apr 1;61(7):2878-84
references: 11306462
references: Cell Death Differ. 2001 May;8(5):477-85
references: 11423908
references: J Cell Sci. 2002 Feb 1;115(Pt 3):599-607
references: 11861766
references: Proc Natl Acad Sci U S A. 2002 Mar 5;99(5):3165-70
references: 11880650
references: Infect Immun. 2002 Nov;70(11):5972-81
references: 12379672
references: Clin Diagn Lab Immunol. 2003 Sep;10(5):903-9
references: 12965925
references: J Biol Chem. 2004 Jan 30;279(5):3578-87
references: 14581476
references: Cell Death Differ. 2004 May;11(5):550-63
references: 14739942
references: Hepatology. 2004 Jul;40(1):185-94
references: 15239102
references: Cytometry A. 2004 Oct;61(2):170-77
references: 15382026
references: J Immunol. 1994 Dec 1;153(11):5141-7
references: 7525724
references: Infect Immun. 1999 Feb;67(2):885-90
references: 9916104
references: Infect Immun. 1999 Sep;67(9):4620-7
references: 10456908
references: J Immunol. 2005 May 15;174(10):6346-56
references: 15879135
references: J Immunol. 2005 Jun 15;174(12):7920-8
references: 15944298
references: Gastroenterology. 2005 Jul;129(1):269-84
references: 16012953
references: Int J Exp Pathol. 2005 Oct;86(5):309-21
references: 16191103
references: J Biol Chem. 2005 Dec 9;280(49):40985-95
references: 16188880
references: J Cancer Res Clin Oncol. 2006 Mar;132(3):171-83
references: 16362335
references: Am J Pathol. 2006 Mar;168(3):847-55
references: 16507900
references: Am J Physiol Gastrointest Liver Physiol. 2006 Jun;290(6):G1339-46
references: 16484678
references: Infect Immun. 2006 Jul;74(7):4339-48
references: 16790808
references: Proc Natl Acad Sci U S A. 2006 Oct 24;103(43):15945-50
references: 17043215
references: Curr Biol. 2006 Nov 7;16(21):2156-60
references: 17084701
references: Curr Biol. 2006 Nov 7;16(21):2161-5
references: 17084702
references: J Biol Chem. 2007 Sep 28;282(39):28960-70
references: 17686764
references: J Immunol. 2007 Oct 15;179(8):5367-77
references: 17911623
references: Int J Mol Med. 2007 Dec;20(6):823-8
references: 17982689
references: Mol Cancer Ther. 2008 Feb;7(2):297-313
references: 18281515
references: Infect Immun. 2008 Jun;76(6):2379-91
references: 18391002