Transcriptional Profiling Suggests T Cells Cluster around Neurons Injected with <named-content content-type="genus-species">Toxoplasma gondii</named-content> Proteins

ABSTRACT Toxoplasma gondii’s tropism for and persistence in the central nervous system (CNS) underlies the symptomatic disease that T. gondii causes in humans. Our recent work has shown that neurons are the primary CNS cell with which Toxoplasma interacts and which it infects in vivo. This predilect...

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Autores principales:	Emily F. Merritt, Hannah J. Johnson, Zhee Sheen Wong, Adam S. Buntzman, Austin C. Conklin, Carla M. Cabral, Casey E. Romanoski, Jon P. Boyle, Anita A. Koshy
Formato:	article
Lenguaje:	EN
Publicado:	American Society for Microbiology 2020
Materias:	RNA-seq Toxoplasma gondii host-pathogen interactions immunology laser capture microdissection neuroscience Microbiology QR1-502
Acceso en línea:	https://doaj.org/article/8ec6668ebe26445abbe5da52c08286cf
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spelling	oai:doaj.org-article:8ec6668ebe26445abbe5da52c08286cf2021-11-15T15:30:58ZTranscriptional Profiling Suggests T Cells Cluster around Neurons Injected with <named-content content-type="genus-species">Toxoplasma gondii</named-content> Proteins10.1128/mSphere.00538-202379-5042https://doaj.org/article/8ec6668ebe26445abbe5da52c08286cf2020-10-01T00:00:00Zhttps://journals.asm.org/doi/10.1128/mSphere.00538-20https://doaj.org/toc/2379-5042ABSTRACT Toxoplasma gondii’s tropism for and persistence in the central nervous system (CNS) underlies the symptomatic disease that T. gondii causes in humans. Our recent work has shown that neurons are the primary CNS cell with which Toxoplasma interacts and which it infects in vivo. This predilection for neurons suggests that T. gondii’s persistence in the CNS depends specifically upon parasite manipulation of the host neurons. Yet, most work on T. gondii-host cell interactions has been done in vitro and in nonneuronal cells. We address this gap by utilizing our T. gondii-Cre system that allows permanent marking and tracking of neurons injected with parasite effector proteins in vivo. Using laser capture microdissection (LCM) and RNA sequencing using RNA-seq, we isolated and transcriptionally profiled T. gondii-injected neurons (TINs), Bystander neurons (nearby non-T. gondii-injected neurons), and neurons from uninfected mice (controls). These profiles show that TIN transcriptomes significantly differ from the transcriptomes of Bystander and control neurons and that much of this difference is driven by increased levels of transcripts from immune cells, especially CD8+ T cells and monocytes. These data suggest that when we used LCM to isolate neurons from infected mice, we also picked up fragments of CD8+ T cells and monocytes clustering in extreme proximity around TINs and, to a lesser extent, Bystander neurons. In addition, we found that T. gondii transcripts were primarily found in the TIN transcriptome, not in the Bystander transcriptome. Collectively, these data suggest that, contrary to common perception, neurons that directly interact with or harbor parasites can be recognized by CD8+ T cells. IMPORTANCE Like other persistent intracellular pathogens, Toxoplasma gondii, a protozoan parasite, has evolved to evade the immune system and establish a chronic infection in specific cells and organs, including neurons in the CNS. Understanding T. gondii’s persistence in neurons holds the potential to identify novel, curative drug targets. The work presented here offers new insights into the neuron-T. gondii interaction in vivo. By transcriptionally profiling neurons manipulated by T. gondii, we unexpectedly revealed that immune cells, and specifically CD8+ T cells, appear to cluster around these neurons, suggesting that CD8+ T cells specifically recognize parasite-manipulated neurons. Such a possibility supports evidence from other labs that questions the long-standing dogma that neurons are often persistently infected because they are not directly recognized by immune cells such as CD8+ T cells. Collectively, these data suggest we reconsider the broader role of neurons in the context of infection and neuroinflammation.Emily F. MerrittHannah J. JohnsonZhee Sheen WongAdam S. BuntzmanAustin C. ConklinCarla M. CabralCasey E. RomanoskiJon P. BoyleAnita A. KoshyAmerican Society for MicrobiologyarticleRNA-seqToxoplasma gondiihost-pathogen interactionsimmunologylaser capture microdissectionneuroscienceMicrobiologyQR1-502ENmSphere, Vol 5, Iss 5 (2020)
institution	DOAJ
collection	DOAJ
language	EN
topic	RNA-seq Toxoplasma gondii host-pathogen interactions immunology laser capture microdissection neuroscience Microbiology QR1-502
spellingShingle	RNA-seq Toxoplasma gondii host-pathogen interactions immunology laser capture microdissection neuroscience Microbiology QR1-502 Emily F. Merritt Hannah J. Johnson Zhee Sheen Wong Adam S. Buntzman Austin C. Conklin Carla M. Cabral Casey E. Romanoski Jon P. Boyle Anita A. Koshy Transcriptional Profiling Suggests T Cells Cluster around Neurons Injected with <named-content content-type="genus-species">Toxoplasma gondii</named-content> Proteins
description	ABSTRACT Toxoplasma gondii’s tropism for and persistence in the central nervous system (CNS) underlies the symptomatic disease that T. gondii causes in humans. Our recent work has shown that neurons are the primary CNS cell with which Toxoplasma interacts and which it infects in vivo. This predilection for neurons suggests that T. gondii’s persistence in the CNS depends specifically upon parasite manipulation of the host neurons. Yet, most work on T. gondii-host cell interactions has been done in vitro and in nonneuronal cells. We address this gap by utilizing our T. gondii-Cre system that allows permanent marking and tracking of neurons injected with parasite effector proteins in vivo. Using laser capture microdissection (LCM) and RNA sequencing using RNA-seq, we isolated and transcriptionally profiled T. gondii-injected neurons (TINs), Bystander neurons (nearby non-T. gondii-injected neurons), and neurons from uninfected mice (controls). These profiles show that TIN transcriptomes significantly differ from the transcriptomes of Bystander and control neurons and that much of this difference is driven by increased levels of transcripts from immune cells, especially CD8+ T cells and monocytes. These data suggest that when we used LCM to isolate neurons from infected mice, we also picked up fragments of CD8+ T cells and monocytes clustering in extreme proximity around TINs and, to a lesser extent, Bystander neurons. In addition, we found that T. gondii transcripts were primarily found in the TIN transcriptome, not in the Bystander transcriptome. Collectively, these data suggest that, contrary to common perception, neurons that directly interact with or harbor parasites can be recognized by CD8+ T cells. IMPORTANCE Like other persistent intracellular pathogens, Toxoplasma gondii, a protozoan parasite, has evolved to evade the immune system and establish a chronic infection in specific cells and organs, including neurons in the CNS. Understanding T. gondii’s persistence in neurons holds the potential to identify novel, curative drug targets. The work presented here offers new insights into the neuron-T. gondii interaction in vivo. By transcriptionally profiling neurons manipulated by T. gondii, we unexpectedly revealed that immune cells, and specifically CD8+ T cells, appear to cluster around these neurons, suggesting that CD8+ T cells specifically recognize parasite-manipulated neurons. Such a possibility supports evidence from other labs that questions the long-standing dogma that neurons are often persistently infected because they are not directly recognized by immune cells such as CD8+ T cells. Collectively, these data suggest we reconsider the broader role of neurons in the context of infection and neuroinflammation.
format	article
author	Emily F. Merritt Hannah J. Johnson Zhee Sheen Wong Adam S. Buntzman Austin C. Conklin Carla M. Cabral Casey E. Romanoski Jon P. Boyle Anita A. Koshy
author_facet	Emily F. Merritt Hannah J. Johnson Zhee Sheen Wong Adam S. Buntzman Austin C. Conklin Carla M. Cabral Casey E. Romanoski Jon P. Boyle Anita A. Koshy
author_sort	Emily F. Merritt
title	Transcriptional Profiling Suggests T Cells Cluster around Neurons Injected with <named-content content-type="genus-species">Toxoplasma gondii</named-content> Proteins
title_short	Transcriptional Profiling Suggests T Cells Cluster around Neurons Injected with <named-content content-type="genus-species">Toxoplasma gondii</named-content> Proteins
title_full	Transcriptional Profiling Suggests T Cells Cluster around Neurons Injected with <named-content content-type="genus-species">Toxoplasma gondii</named-content> Proteins
title_fullStr	Transcriptional Profiling Suggests T Cells Cluster around Neurons Injected with <named-content content-type="genus-species">Toxoplasma gondii</named-content> Proteins
title_full_unstemmed	Transcriptional Profiling Suggests T Cells Cluster around Neurons Injected with <named-content content-type="genus-species">Toxoplasma gondii</named-content> Proteins
title_sort	transcriptional profiling suggests t cells cluster around neurons injected with <named-content content-type="genus-species">toxoplasma gondii</named-content> proteins
publisher	American Society for Microbiology
publishDate	2020
url	https://doaj.org/article/8ec6668ebe26445abbe5da52c08286cf
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Transcriptional Profiling Suggests T Cells Cluster around Neurons Injected with <named-content content-type="genus-species">Toxoplasma gondii</named-content> Proteins

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