Dynamics of <named-content content-type="genus-species">Mycobacterium tuberculosis</named-content> Ag85B Revealed by a Sensitive Enzyme-Linked Immunosorbent Assay

ABSTRACT Secretion of specific proteins contributes to pathogenesis and immune responses in tuberculosis and other bacterial infections, yet the kinetics of protein secretion and fate of secreted proteins in vivo are poorly understood. We generated new monoclonal antibodies that recognize the Mycoba...

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Autores principales:	Joel D. Ernst, Amber Cornelius, Miriam Bolz
Formato:	article
Lenguaje:	EN
Publicado:	American Society for Microbiology 2019
Materias:	Ag85B ELISA bacterial antigen bacterial protein secretion monoclonal antibodies tuberculosis Microbiology QR1-502
Acceso en línea:	https://doaj.org/article/173c39e1cc6e4be997263edbf09e92f2
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spelling	oai:doaj.org-article:173c39e1cc6e4be997263edbf09e92f22021-11-15T15:55:24ZDynamics of <named-content content-type="genus-species">Mycobacterium tuberculosis</named-content> Ag85B Revealed by a Sensitive Enzyme-Linked Immunosorbent Assay10.1128/mBio.00611-192150-7511https://doaj.org/article/173c39e1cc6e4be997263edbf09e92f22019-04-01T00:00:00Zhttps://journals.asm.org/doi/10.1128/mBio.00611-19https://doaj.org/toc/2150-7511ABSTRACT Secretion of specific proteins contributes to pathogenesis and immune responses in tuberculosis and other bacterial infections, yet the kinetics of protein secretion and fate of secreted proteins in vivo are poorly understood. We generated new monoclonal antibodies that recognize the Mycobacterium tuberculosis secreted protein Ag85B and used them to establish and characterize a sensitive enzyme-linked immunosorbent assay (ELISA) to quantitate Ag85B in samples generated in vitro and in vivo. We found that nutritional or culture conditions had little impact on the secretion of Ag85B and that there is considerable variation in Ag85B secretion by distinct strains in the M. tuberculosis complex: compared with the commonly used H37Rv strain (lineage 4), Mycobacterium africanum (lineage 6) secretes less Ag85B, and two strains from lineage 2 secrete more Ag85B. We also used the ELISA to determine that the rate of secretion of Ag85B is 10- to 100-fold lower than that of proteins secreted by Gram-negative and Gram-positive bacteria, respectively. ELISA quantitation of Ag85B in lung homogenates of M. tuberculosis H37Rv-infected mice revealed that although Ag85B accumulates in the lungs as the bacterial population expands, the amount of Ag85B per bacterium decreases nearly 10,000-fold at later stages of infection, coincident with the development of T cell responses and arrest of bacterial population growth. These results indicate that bacterial protein secretion in vivo is dynamic and regulated, and quantitation of secreted bacterial proteins can contribute to the understanding of pathogenesis and immunity in tuberculosis and other infections. IMPORTANCE Bacterial protein secretion contributes to host-pathogen interactions, yet the process and consequences of bacterial protein secretion during infection are poorly understood. We developed a sensitive ELISA to quantitate a protein (termed Ag85B) secreted by M. tuberculosis and used it to find that Ag85B secretion occurs with slower kinetics than for proteins secreted by Gram-positive and Gram-negative bacteria and that accumulation of Ag85B in the lungs is markedly regulated as a function of the bacterial population density. Our results demonstrate that quantitation of bacterial proteins during infection can reveal novel insights into host-pathogen interactions.Joel D. ErnstAmber CorneliusMiriam BolzAmerican Society for MicrobiologyarticleAg85BELISAbacterial antigenbacterial protein secretionmonoclonal antibodiestuberculosisMicrobiologyQR1-502ENmBio, Vol 10, Iss 2 (2019)
institution	DOAJ
collection	DOAJ
language	EN
topic	Ag85B ELISA bacterial antigen bacterial protein secretion monoclonal antibodies tuberculosis Microbiology QR1-502
spellingShingle	Ag85B ELISA bacterial antigen bacterial protein secretion monoclonal antibodies tuberculosis Microbiology QR1-502 Joel D. Ernst Amber Cornelius Miriam Bolz Dynamics of <named-content content-type="genus-species">Mycobacterium tuberculosis</named-content> Ag85B Revealed by a Sensitive Enzyme-Linked Immunosorbent Assay
description	ABSTRACT Secretion of specific proteins contributes to pathogenesis and immune responses in tuberculosis and other bacterial infections, yet the kinetics of protein secretion and fate of secreted proteins in vivo are poorly understood. We generated new monoclonal antibodies that recognize the Mycobacterium tuberculosis secreted protein Ag85B and used them to establish and characterize a sensitive enzyme-linked immunosorbent assay (ELISA) to quantitate Ag85B in samples generated in vitro and in vivo. We found that nutritional or culture conditions had little impact on the secretion of Ag85B and that there is considerable variation in Ag85B secretion by distinct strains in the M. tuberculosis complex: compared with the commonly used H37Rv strain (lineage 4), Mycobacterium africanum (lineage 6) secretes less Ag85B, and two strains from lineage 2 secrete more Ag85B. We also used the ELISA to determine that the rate of secretion of Ag85B is 10- to 100-fold lower than that of proteins secreted by Gram-negative and Gram-positive bacteria, respectively. ELISA quantitation of Ag85B in lung homogenates of M. tuberculosis H37Rv-infected mice revealed that although Ag85B accumulates in the lungs as the bacterial population expands, the amount of Ag85B per bacterium decreases nearly 10,000-fold at later stages of infection, coincident with the development of T cell responses and arrest of bacterial population growth. These results indicate that bacterial protein secretion in vivo is dynamic and regulated, and quantitation of secreted bacterial proteins can contribute to the understanding of pathogenesis and immunity in tuberculosis and other infections. IMPORTANCE Bacterial protein secretion contributes to host-pathogen interactions, yet the process and consequences of bacterial protein secretion during infection are poorly understood. We developed a sensitive ELISA to quantitate a protein (termed Ag85B) secreted by M. tuberculosis and used it to find that Ag85B secretion occurs with slower kinetics than for proteins secreted by Gram-positive and Gram-negative bacteria and that accumulation of Ag85B in the lungs is markedly regulated as a function of the bacterial population density. Our results demonstrate that quantitation of bacterial proteins during infection can reveal novel insights into host-pathogen interactions.
format	article
author	Joel D. Ernst Amber Cornelius Miriam Bolz
author_facet	Joel D. Ernst Amber Cornelius Miriam Bolz
author_sort	Joel D. Ernst
title	Dynamics of <named-content content-type="genus-species">Mycobacterium tuberculosis</named-content> Ag85B Revealed by a Sensitive Enzyme-Linked Immunosorbent Assay
title_short	Dynamics of <named-content content-type="genus-species">Mycobacterium tuberculosis</named-content> Ag85B Revealed by a Sensitive Enzyme-Linked Immunosorbent Assay
title_full	Dynamics of <named-content content-type="genus-species">Mycobacterium tuberculosis</named-content> Ag85B Revealed by a Sensitive Enzyme-Linked Immunosorbent Assay
title_fullStr	Dynamics of <named-content content-type="genus-species">Mycobacterium tuberculosis</named-content> Ag85B Revealed by a Sensitive Enzyme-Linked Immunosorbent Assay
title_full_unstemmed	Dynamics of <named-content content-type="genus-species">Mycobacterium tuberculosis</named-content> Ag85B Revealed by a Sensitive Enzyme-Linked Immunosorbent Assay
title_sort	dynamics of <named-content content-type="genus-species">mycobacterium tuberculosis</named-content> ag85b revealed by a sensitive enzyme-linked immunosorbent assay
publisher	American Society for Microbiology
publishDate	2019
url	https://doaj.org/article/173c39e1cc6e4be997263edbf09e92f2
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Dynamics of <named-content content-type="genus-species">Mycobacterium tuberculosis</named-content> Ag85B Revealed by a Sensitive Enzyme-Linked Immunosorbent Assay

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