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Single-Cell Sequencing Confirms Transcripts and V(H)DJ(H) Rearrangements of Immunoglobulin Genes in Human Podocytes

Most glomerular diseases are associated with inflammation caused by deposited pathogenic immunoglobulins (Igs), which are believed to be produced by B cells. However, our previous study indicated that the human podocyte cell line can produce IgG. In this study, we aimed to confirm the transcripts an...

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Autores principales: Deng, Zhenling, Yan, Huige, Shi, Zhan, Tian, Xinyu, Cui, Zhuan, Sun, Yingchun, Wang, Song, Zheng, Danxia, Qiu, Xiaoyan, Wang, Yue
Formato: Online Artículo Texto
Lenguaje:English
Publicado: MDPI 2021
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC8064494/
https://www.ncbi.nlm.nih.gov/pubmed/33806147
http://dx.doi.org/10.3390/genes12040472
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author Deng, Zhenling
Yan, Huige
Shi, Zhan
Tian, Xinyu
Cui, Zhuan
Sun, Yingchun
Wang, Song
Zheng, Danxia
Qiu, Xiaoyan
Wang, Yue
author_facet Deng, Zhenling
Yan, Huige
Shi, Zhan
Tian, Xinyu
Cui, Zhuan
Sun, Yingchun
Wang, Song
Zheng, Danxia
Qiu, Xiaoyan
Wang, Yue
author_sort Deng, Zhenling
collection PubMed
description Most glomerular diseases are associated with inflammation caused by deposited pathogenic immunoglobulins (Igs), which are believed to be produced by B cells. However, our previous study indicated that the human podocyte cell line can produce IgG. In this study, we aimed to confirm the transcripts and characterize the repertoires of Igs in primary podocytes at single cell level. First, single-cell RNA sequencing of cell suspensions from “normal” kidney cortexes by a 10xGenomics Chromium system detected Ig transcripts in 7/360 podocytes and Ig gene segments in 106/360 podocytes. Then, we combined nested PCR with Sanger sequencing to detect the transcripts and characterize the repertoires of Igs in 48 single podocytes and found that five classes of Ig heavy chains were amplified in podocytes. Four-hundred and twenty-nine V(H)DJ(H) rearrangement sequences were analyzed; podocyte-derived Igs exhibited classic V(H)DJ(H) rearrangements with nucleotide additions and somatic hypermutations, biased VH1 usage and restricted diversity. Moreover, compared with the podocytes from healthy control that usually expressed one class of Ig and one V(H)DJ(H) pattern, podocytes from patients expressed more classes of Ig, V(H)DJ(H) patterns and somatic hypermutations. These findings suggested that podocytes can express Igs in normal condition and increase diversity in pathological situations.
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spelling pubmed-80644942021-04-24 Single-Cell Sequencing Confirms Transcripts and V(H)DJ(H) Rearrangements of Immunoglobulin Genes in Human Podocytes Deng, Zhenling Yan, Huige Shi, Zhan Tian, Xinyu Cui, Zhuan Sun, Yingchun Wang, Song Zheng, Danxia Qiu, Xiaoyan Wang, Yue Genes (Basel) Article Most glomerular diseases are associated with inflammation caused by deposited pathogenic immunoglobulins (Igs), which are believed to be produced by B cells. However, our previous study indicated that the human podocyte cell line can produce IgG. In this study, we aimed to confirm the transcripts and characterize the repertoires of Igs in primary podocytes at single cell level. First, single-cell RNA sequencing of cell suspensions from “normal” kidney cortexes by a 10xGenomics Chromium system detected Ig transcripts in 7/360 podocytes and Ig gene segments in 106/360 podocytes. Then, we combined nested PCR with Sanger sequencing to detect the transcripts and characterize the repertoires of Igs in 48 single podocytes and found that five classes of Ig heavy chains were amplified in podocytes. Four-hundred and twenty-nine V(H)DJ(H) rearrangement sequences were analyzed; podocyte-derived Igs exhibited classic V(H)DJ(H) rearrangements with nucleotide additions and somatic hypermutations, biased VH1 usage and restricted diversity. Moreover, compared with the podocytes from healthy control that usually expressed one class of Ig and one V(H)DJ(H) pattern, podocytes from patients expressed more classes of Ig, V(H)DJ(H) patterns and somatic hypermutations. These findings suggested that podocytes can express Igs in normal condition and increase diversity in pathological situations. MDPI 2021-03-25 /pmc/articles/PMC8064494/ /pubmed/33806147 http://dx.doi.org/10.3390/genes12040472 Text en © 2021 by the authors. https://creativecommons.org/licenses/by/4.0/Licensee MDPI, Basel, Switzerland. This article is an open access article distributed under the terms and conditions of the Creative Commons Attribution (CC BY) license (http://creativecommons.org/licenses/by/4.0/ (https://creativecommons.org/licenses/by/4.0/) ).
spellingShingle Article
Deng, Zhenling
Yan, Huige
Shi, Zhan
Tian, Xinyu
Cui, Zhuan
Sun, Yingchun
Wang, Song
Zheng, Danxia
Qiu, Xiaoyan
Wang, Yue
Single-Cell Sequencing Confirms Transcripts and V(H)DJ(H) Rearrangements of Immunoglobulin Genes in Human Podocytes
title Single-Cell Sequencing Confirms Transcripts and V(H)DJ(H) Rearrangements of Immunoglobulin Genes in Human Podocytes
title_full Single-Cell Sequencing Confirms Transcripts and V(H)DJ(H) Rearrangements of Immunoglobulin Genes in Human Podocytes
title_fullStr Single-Cell Sequencing Confirms Transcripts and V(H)DJ(H) Rearrangements of Immunoglobulin Genes in Human Podocytes
title_full_unstemmed Single-Cell Sequencing Confirms Transcripts and V(H)DJ(H) Rearrangements of Immunoglobulin Genes in Human Podocytes
title_short Single-Cell Sequencing Confirms Transcripts and V(H)DJ(H) Rearrangements of Immunoglobulin Genes in Human Podocytes
title_sort single-cell sequencing confirms transcripts and v(h)dj(h) rearrangements of immunoglobulin genes in human podocytes
topic Article
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC8064494/
https://www.ncbi.nlm.nih.gov/pubmed/33806147
http://dx.doi.org/10.3390/genes12040472
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