Immunomagnetic B cell isolation as a tool to study blood cell subsets and enrich B cell transcripts

Abstract Objective Transcriptional profiling of immune cells is an indispensable tool in biomedical research; however, heterogenous sample types routinely used in transcriptomic studies may mask important cell type-specific transcriptional differences. Techniques to isolate desired cell types are us...

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Autores principales:	Amanda N. Henning, Daniel Green, Ryan Baumann, Patrick Grandinetti, Steven L. Highfill, Huizhi Zhou, Valeria De Giorgi
Formato:	article
Lenguaje:	EN
Publicado:	BMC 2021
Materias:	RNA-sequencing B cells Differential gene expression Gene signature Medicine R Biology (General) QH301-705.5 Science (General) Q1-390
Acceso en línea:	https://doaj.org/article/dca354737e5b4efc8d253cdf48cfc1a0
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spelling	oai:doaj.org-article:dca354737e5b4efc8d253cdf48cfc1a02021-11-21T12:28:05ZImmunomagnetic B cell isolation as a tool to study blood cell subsets and enrich B cell transcripts10.1186/s13104-021-05833-z1756-0500https://doaj.org/article/dca354737e5b4efc8d253cdf48cfc1a02021-11-01T00:00:00Zhttps://doi.org/10.1186/s13104-021-05833-zhttps://doaj.org/toc/1756-0500Abstract Objective Transcriptional profiling of immune cells is an indispensable tool in biomedical research; however, heterogenous sample types routinely used in transcriptomic studies may mask important cell type-specific transcriptional differences. Techniques to isolate desired cell types are used to overcome this limitation. We sought to evaluate the use of immunomagnetic B cell isolation on RNA quality and transcriptional output. Additionally, we aimed to develop a B cell gene signature representative of a freshly isolated B cell population to be used as a tool to verify isolation efficacy and to provide a transcriptional standard for evaluating maintenance or deviation from traditional B cell identity. Results We found RNA quality and RNA-sequencing output to be comparable between donor-matched PBMC, whole blood, and B cells following negative selection by immunomagnetic B cell isolation. Transcriptional analysis enabled the development of an 85 gene B cell signature. This signature effectively clustered isolated B cells from heterogeneous sample types in our study and naïve and memory B cells when applied to transcriptional data from a published source. Additionally, by identifying B cell signature genes whose functional role in B cells is currently unknown, our gene signature has uncovered areas for future investigation.Amanda N. HenningDaniel GreenRyan BaumannPatrick GrandinettiSteven L. HighfillHuizhi ZhouValeria De GiorgiBMCarticleRNA-sequencingB cellsDifferential gene expressionGene signatureMedicineRBiology (General)QH301-705.5Science (General)Q1-390ENBMC Research Notes, Vol 14, Iss 1, Pp 1-7 (2021)
institution	DOAJ
collection	DOAJ
language	EN
topic	RNA-sequencing B cells Differential gene expression Gene signature Medicine R Biology (General) QH301-705.5 Science (General) Q1-390
spellingShingle	RNA-sequencing B cells Differential gene expression Gene signature Medicine R Biology (General) QH301-705.5 Science (General) Q1-390 Amanda N. Henning Daniel Green Ryan Baumann Patrick Grandinetti Steven L. Highfill Huizhi Zhou Valeria De Giorgi Immunomagnetic B cell isolation as a tool to study blood cell subsets and enrich B cell transcripts
description	Abstract Objective Transcriptional profiling of immune cells is an indispensable tool in biomedical research; however, heterogenous sample types routinely used in transcriptomic studies may mask important cell type-specific transcriptional differences. Techniques to isolate desired cell types are used to overcome this limitation. We sought to evaluate the use of immunomagnetic B cell isolation on RNA quality and transcriptional output. Additionally, we aimed to develop a B cell gene signature representative of a freshly isolated B cell population to be used as a tool to verify isolation efficacy and to provide a transcriptional standard for evaluating maintenance or deviation from traditional B cell identity. Results We found RNA quality and RNA-sequencing output to be comparable between donor-matched PBMC, whole blood, and B cells following negative selection by immunomagnetic B cell isolation. Transcriptional analysis enabled the development of an 85 gene B cell signature. This signature effectively clustered isolated B cells from heterogeneous sample types in our study and naïve and memory B cells when applied to transcriptional data from a published source. Additionally, by identifying B cell signature genes whose functional role in B cells is currently unknown, our gene signature has uncovered areas for future investigation.
format	article
author	Amanda N. Henning Daniel Green Ryan Baumann Patrick Grandinetti Steven L. Highfill Huizhi Zhou Valeria De Giorgi
author_facet	Amanda N. Henning Daniel Green Ryan Baumann Patrick Grandinetti Steven L. Highfill Huizhi Zhou Valeria De Giorgi
author_sort	Amanda N. Henning
title	Immunomagnetic B cell isolation as a tool to study blood cell subsets and enrich B cell transcripts
title_short	Immunomagnetic B cell isolation as a tool to study blood cell subsets and enrich B cell transcripts
title_full	Immunomagnetic B cell isolation as a tool to study blood cell subsets and enrich B cell transcripts
title_fullStr	Immunomagnetic B cell isolation as a tool to study blood cell subsets and enrich B cell transcripts
title_full_unstemmed	Immunomagnetic B cell isolation as a tool to study blood cell subsets and enrich B cell transcripts
title_sort	immunomagnetic b cell isolation as a tool to study blood cell subsets and enrich b cell transcripts
publisher	BMC
publishDate	2021
url	https://doaj.org/article/dca354737e5b4efc8d253cdf48cfc1a0
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Immunomagnetic B cell isolation as a tool to study blood cell subsets and enrich B cell transcripts

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