Transcriptomic and Proteomic Approaches to Finding Novel Diagnostic and Immunogenic Candidates in <italic toggle="yes">Pneumocystis</italic>

ABSTRACT Pneumocystis pneumonia is the most common serious opportunistic infection in patients with HIV/AIDS. Furthermore, Pneumocystis pneumonia is a feared complication of the immunosuppressive drug regimens used to treat autoimmunity, malignancy, and posttransplantation rejection. With an increas...

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Autores principales:	Taylor Eddens, Waleed Elsegeiny, David Ricks, Meagan Goodwin, William T. Horne, Mingquan Zheng, Jay K. Kolls
Formato:	article
Lenguaje:	EN
Publicado:	American Society for Microbiology 2019
Materias:	Pneumocystis diagnostics life cycle transcriptomics vaccines Microbiology QR1-502
Acceso en línea:	https://doaj.org/article/7875506580b74fc792633149e3c8237d
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spelling	oai:doaj.org-article:7875506580b74fc792633149e3c8237d2021-11-15T15:27:32ZTranscriptomic and Proteomic Approaches to Finding Novel Diagnostic and Immunogenic Candidates in <italic toggle="yes">Pneumocystis</italic>10.1128/mSphere.00488-192379-5042https://doaj.org/article/7875506580b74fc792633149e3c8237d2019-10-01T00:00:00Zhttps://journals.asm.org/doi/10.1128/mSphere.00488-19https://doaj.org/toc/2379-5042ABSTRACT Pneumocystis pneumonia is the most common serious opportunistic infection in patients with HIV/AIDS. Furthermore, Pneumocystis pneumonia is a feared complication of the immunosuppressive drug regimens used to treat autoimmunity, malignancy, and posttransplantation rejection. With an increasing at-risk population, there is a strong need for novel approaches to discover diagnostic and vaccine targets. There are multiple challenges to finding these targets, however. First, Pneumocystis has a largely unannotated genome. To address this, we evaluated each protein encoded within the Pneumocystis genome by comparisons to proteins encoded within the genomes of other fungi using NCBI BLAST. Second, Pneumocystis relies on a multiphasic life cycle, as both the transmissible form (the ascus) and the replicative form (the trophozoite [troph]) reside within the alveolar space of the host. To that end, we purified asci and trophs from Pneumocystis murina and utilized transcriptomics to identify differentially regulated genes. Two such genes, Arp9 and Sp, are differentially regulated in the ascus and the troph, respectively, and can be utilized to characterize the state of the Pneumocystis life cycle in vivo. Gsc1, encoding a β-1,3-glucan synthase with a large extracellular domain previously identified using surface proteomics, was more highly expressed on the ascus form of Pneumocystis. GSC-1 ectodomain immunization generated a strong antibody response that demonstrated the ability to recognize the surface of the Pneumocystis asci. GSC-1 ectodomain immunization was also capable of reducing ascus burden following primary challenge with Pneumocystis murina. Finally, mice immunized with the GSC-1 ectodomain had limited fungal burden following natural transmission of Pneumocystis using a cohousing model. IMPORTANCE The current report enhances our understanding of Pneumocystis biology in a number of ways. First, the current study provided a preliminary annotation of the Pneumocystis murina genome, addressing a long-standing issue in the field. Second, this study validated two novel transcripts enriched in the two predominant life forms of Pneumocystis. These findings allow better characterization of the Pneumocystis life cycle in vivo and could be valuable diagnostic tools. Furthermore, this study outlined a novel pipeline of -omics techniques capable of revealing novel antigens (e.g., GSC-1) for the development of vaccines against Pneumocystis.Taylor EddensWaleed ElsegeinyDavid RicksMeagan GoodwinWilliam T. HorneMingquan ZhengJay K. KollsAmerican Society for MicrobiologyarticlePneumocystisdiagnosticslife cycletranscriptomicsvaccinesMicrobiologyQR1-502ENmSphere, Vol 4, Iss 5 (2019)
institution	DOAJ
collection	DOAJ
language	EN
topic	Pneumocystis diagnostics life cycle transcriptomics vaccines Microbiology QR1-502
spellingShingle	Pneumocystis diagnostics life cycle transcriptomics vaccines Microbiology QR1-502 Taylor Eddens Waleed Elsegeiny David Ricks Meagan Goodwin William T. Horne Mingquan Zheng Jay K. Kolls Transcriptomic and Proteomic Approaches to Finding Novel Diagnostic and Immunogenic Candidates in <italic toggle="yes">Pneumocystis</italic>
description	ABSTRACT Pneumocystis pneumonia is the most common serious opportunistic infection in patients with HIV/AIDS. Furthermore, Pneumocystis pneumonia is a feared complication of the immunosuppressive drug regimens used to treat autoimmunity, malignancy, and posttransplantation rejection. With an increasing at-risk population, there is a strong need for novel approaches to discover diagnostic and vaccine targets. There are multiple challenges to finding these targets, however. First, Pneumocystis has a largely unannotated genome. To address this, we evaluated each protein encoded within the Pneumocystis genome by comparisons to proteins encoded within the genomes of other fungi using NCBI BLAST. Second, Pneumocystis relies on a multiphasic life cycle, as both the transmissible form (the ascus) and the replicative form (the trophozoite [troph]) reside within the alveolar space of the host. To that end, we purified asci and trophs from Pneumocystis murina and utilized transcriptomics to identify differentially regulated genes. Two such genes, Arp9 and Sp, are differentially regulated in the ascus and the troph, respectively, and can be utilized to characterize the state of the Pneumocystis life cycle in vivo. Gsc1, encoding a β-1,3-glucan synthase with a large extracellular domain previously identified using surface proteomics, was more highly expressed on the ascus form of Pneumocystis. GSC-1 ectodomain immunization generated a strong antibody response that demonstrated the ability to recognize the surface of the Pneumocystis asci. GSC-1 ectodomain immunization was also capable of reducing ascus burden following primary challenge with Pneumocystis murina. Finally, mice immunized with the GSC-1 ectodomain had limited fungal burden following natural transmission of Pneumocystis using a cohousing model. IMPORTANCE The current report enhances our understanding of Pneumocystis biology in a number of ways. First, the current study provided a preliminary annotation of the Pneumocystis murina genome, addressing a long-standing issue in the field. Second, this study validated two novel transcripts enriched in the two predominant life forms of Pneumocystis. These findings allow better characterization of the Pneumocystis life cycle in vivo and could be valuable diagnostic tools. Furthermore, this study outlined a novel pipeline of -omics techniques capable of revealing novel antigens (e.g., GSC-1) for the development of vaccines against Pneumocystis.
format	article
author	Taylor Eddens Waleed Elsegeiny David Ricks Meagan Goodwin William T. Horne Mingquan Zheng Jay K. Kolls
author_facet	Taylor Eddens Waleed Elsegeiny David Ricks Meagan Goodwin William T. Horne Mingquan Zheng Jay K. Kolls
author_sort	Taylor Eddens
title	Transcriptomic and Proteomic Approaches to Finding Novel Diagnostic and Immunogenic Candidates in <italic toggle="yes">Pneumocystis</italic>
title_short	Transcriptomic and Proteomic Approaches to Finding Novel Diagnostic and Immunogenic Candidates in <italic toggle="yes">Pneumocystis</italic>
title_full	Transcriptomic and Proteomic Approaches to Finding Novel Diagnostic and Immunogenic Candidates in <italic toggle="yes">Pneumocystis</italic>
title_fullStr	Transcriptomic and Proteomic Approaches to Finding Novel Diagnostic and Immunogenic Candidates in <italic toggle="yes">Pneumocystis</italic>
title_full_unstemmed	Transcriptomic and Proteomic Approaches to Finding Novel Diagnostic and Immunogenic Candidates in <italic toggle="yes">Pneumocystis</italic>
title_sort	transcriptomic and proteomic approaches to finding novel diagnostic and immunogenic candidates in <italic toggle="yes">pneumocystis</italic>
publisher	American Society for Microbiology
publishDate	2019
url	https://doaj.org/article/7875506580b74fc792633149e3c8237d
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Transcriptomic and Proteomic Approaches to Finding Novel Diagnostic and Immunogenic Candidates in <italic toggle="yes">Pneumocystis</italic>

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