PMID- 32853276 OWN - NLM STAT- MEDLINE DCOM- 20200922 LR - 20220202 IS - 1553-7374 (Electronic) IS - 1553-7366 (Print) IS - 1553-7366 (Linking) VI - 16 IP - 8 DP - 2020 Aug TI - Naive CD8 T cell IFNgamma responses to a vacuolar antigen are regulated by an inflammasome-independent NLRP3 pathway and Toxoplasma gondii ROP5. PG - e1008327 LID - 10.1371/journal.ppat.1008327 [doi] LID - e1008327 AB - Host resistance to Toxoplasma gondii relies on CD8 T cell IFNgamma responses, which if modulated by the host or parasite could influence chronic infection and parasite transmission between hosts. Since host-parasite interactions that govern this response are not fully elucidated, we investigated requirements for eliciting naive CD8 T cell IFNgamma responses to a vacuolar resident antigen of T. gondii, TGD057. Naive TGD057 antigen-specific CD8 T cells (T57) were isolated from transnuclear mice and responded to parasite-infected bone marrow-derived macrophages (BMDMs) in an antigen-dependent manner, first by producing IL-2 and then IFNgamma. T57 IFNgamma responses to TGD057 were independent of the parasite's protein export machinery ASP5 and MYR1. Instead, host immunity pathways downstream of the regulatory Immunity-Related GTPases (IRG), including partial dependence on Guanylate-Binding Proteins, are required. Multiple T. gondii ROP5 isoforms and allele types, including 'avirulent' ROP5A from clade A and D parasite strains, were able to suppress CD8 T cell IFNgamma responses to parasite-infected BMDMs. Phenotypic variance between clades B, C, D, F, and A strains suggest T57 IFNgamma differentiation occurs independently of parasite virulence or any known IRG-ROP5 interaction. Consistent with this, removal of ROP5 is not enough to elicit maximal CD8 T cell IFNgamma production to parasite-infected cells. Instead, macrophage expression of the pathogen sensors, NLRP3 and to a large extent NLRP1, were absolute requirements. Other members of the conventional inflammasome cascade are only partially required, as revealed by decreased but not abrogated T57 IFNgamma responses to parasite-infected ASC, caspase-1/11, and gasdermin D deficient cells. Moreover, IFNgamma production was only partially reduced in the absence of IL-12, IL-18 or IL-1R signaling. In summary, T. gondii effectors and host machinery that modulate parasitophorous vacuolar membranes, as well as NLR-dependent but inflammasome-independent pathways, determine the full commitment of CD8 T cells IFNgamma responses to a vacuolar antigen. FAU - Kongsomboonvech, Angel K AU - Kongsomboonvech AK AD - Department of Molecular and Cell Biology, University of California, Merced, Merced, California, United States of America. FAU - Rodriguez, Felipe AU - Rodriguez F AD - Department of Molecular and Cell Biology, University of California, Merced, Merced, California, United States of America. FAU - Diep, Anh L AU - Diep AL AUID- ORCID: 0000-0003-0217-324X AD - Department of Molecular and Cell Biology, University of California, Merced, Merced, California, United States of America. FAU - Justice, Brandon M AU - Justice BM AUID- ORCID: 0000-0003-1197-1171 AD - Department of Molecular and Cell Biology, University of California, Merced, Merced, California, United States of America. FAU - Castallanos, Brayan E AU - Castallanos BE AUID- ORCID: 0000-0003-1397-7200 AD - Department of Molecular and Cell Biology, University of California, Merced, Merced, California, United States of America. FAU - Camejo, Ana AU - Camejo A AD - Department of Biology, Massachusetts Institute of Technology, Cambridge, Massachusetts, United States of America. FAU - Mukhopadhyay, Debanjan AU - Mukhopadhyay D AUID- ORCID: 0000-0002-8921-4106 AD - Department of Pathology, Microbiology and Immunology, School of Veterinary Medicine, University of California, Davis, Davis, California, United States of America. FAU - Taylor, Gregory A AU - Taylor GA AUID- ORCID: 0000-0003-1966-0917 AD - Departments of Medicine; Molecular Genetics and Microbiology; and Immunology; and Center for the Study of Aging and Human Development, Duke University Medical Center, Durham, North Carolina, United States of America. AD - Geriatric Research, Education, and Clinical Center, Durham VA Health Care System, Durham, North Carolina, United States of America. FAU - Yamamoto, Masahiro AU - Yamamoto M AUID- ORCID: 0000-0002-6821-2785 AD - Department of Immunoparasitology, Research Institute for Microbial Diseases, Osaka University, Osaka, Japan. FAU - Saeij, Jeroen P J AU - Saeij JPJ AUID- ORCID: 0000-0003-0289-7109 AD - Department of Biology, Massachusetts Institute of Technology, Cambridge, Massachusetts, United States of America. AD - Department of Pathology, Microbiology and Immunology, School of Veterinary Medicine, University of California, Davis, Davis, California, United States of America. FAU - Reese, Michael L AU - Reese ML AD - Department of Pharmacology, University of Texas, Southwestern Medical Center, Dallas, Texas, United States of America. FAU - Jensen, Kirk D C AU - Jensen KDC AUID- ORCID: 0000-0003-0707-964X AD - Department of Molecular and Cell Biology, University of California, Merced, Merced, California, United States of America. AD - Health Sciences Research Institute, University of California, Merced, Merced, California, United States of America. LA - eng GR - R01 AI145929/AI/NIAID NIH HHS/United States GR - R15 AI131027/AI/NIAID NIH HHS/United States GR - T32 DK098132/DK/NIDDK NIH HHS/United States GR - R01 AI080621/AI/NIAID NIH HHS/United States PT - Journal Article PT - Research Support, N.I.H., Extramural PT - Research Support, Non-U.S. Gov't PT - Research Support, U.S. Gov't, Non-P.H.S. DEP - 20200827 PL - United States TA - PLoS Pathog JT - PLoS pathogens JID - 101238921 RN - 0 (Inflammasomes) RN - 0 (NLR Family, Pyrin Domain-Containing 3 Protein) RN - 0 (Nlrp3 protein, mouse) RN - 0 (Protozoan Proteins) RN - 82115-62-6 (Interferon-gamma) SB - IM MH - Animals MH - CD8-Positive T-Lymphocytes/*immunology/parasitology MH - Female MH - Inflammasomes/*immunology MH - Interferon-gamma/*metabolism MH - Macrophages/immunology/parasitology MH - Mice MH - NLR Family, Pyrin Domain-Containing 3 Protein/genetics/*metabolism MH - Protozoan Proteins/genetics/*metabolism MH - *Signal Transduction MH - Toxoplasma/*immunology MH - Toxoplasmosis, Animal/*immunology/parasitology MH - Vacuoles/immunology/metabolism/parasitology MH - Virulence/immunology PMC - PMC7480859 COIS- The authors have declared that no competing interests exist. EDAT- 2020/08/28 06:00 MHDA- 2020/09/23 06:00 PMCR- 2020/08/27 CRDT- 2020/08/28 06:00 PHST- 2020/01/13 00:00 [received] PHST- 2020/07/05 00:00 [accepted] PHST- 2020/09/09 00:00 [revised] PHST- 2020/08/28 06:00 [pubmed] PHST- 2020/09/23 06:00 [medline] PHST- 2020/08/28 06:00 [entrez] PHST- 2020/08/27 00:00 [pmc-release] AID - PPATHOGENS-D-20-00067 [pii] AID - 10.1371/journal.ppat.1008327 [doi] PST - epublish SO - PLoS Pathog. 2020 Aug 27;16(8):e1008327. doi: 10.1371/journal.ppat.1008327. eCollection 2020 Aug.