Transcriptional profiling of PBMCs unravels B cell mediated immunopathogenic imprints of HCV vasculitis.
B cell depletion therapy using rituximab has been shown to be effective in achieving remission in patients with HCV-mixed cryoglobulinemic (MC) vasculitis. Previously, we have demonstrated abnormalities in peripheral immune cells involving neutrophils, chemotaxis, and innate immune activation among...
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doaj-58416fd527b5414892ec74a72ddcb0832020-11-25T01:21:31ZengPublic Library of Science (PLoS)PLoS ONE1932-62032017-01-011212e018831410.1371/journal.pone.0188314Transcriptional profiling of PBMCs unravels B cell mediated immunopathogenic imprints of HCV vasculitis.Emily ComstockCheol-Woo KimAlison MurphyBenjamin EmmanuelXi ZhangMichael SnellerBhawna PooniaShyamasundaran KottililB cell depletion therapy using rituximab has been shown to be effective in achieving remission in patients with HCV-mixed cryoglobulinemic (MC) vasculitis. Previously, we have demonstrated abnormalities in peripheral immune cells involving neutrophils, chemotaxis, and innate immune activation among patients with HCV-MC vasculitis when compared to HCV patients without vasculitis. In this study, we evaluated the effect of B cell depletion therapy on transcriptional profiles of peripheral blood mononuclear cells before and after riruximab therapy, in order to unravel the pathogenic mechanism involved in HCV-MC vasculitis induced by abnormal B cell proliferation. DNA microarray analysis was performed using RNA from PBMCs from seven patients with HCV-MC vasculitis and seven normal volunteers. DNA was hybridized to Affymetrix U133A chips. After normalization, differentially expressed gene list with treatment was generated using partitional clustering. RT-PCR, flow cytometry, and enzyme immunoassay (EIA) was used to validate DNA microarray findings. Differentially expressed genes included B cells and non-B cell genes. Validation of genes using purified cell subsets demonstrated distinct effect of B cell depletion therapy on non-B cells, such as monocytes, T cells, and NK cells. Notably, B lymphocyte stimulator (BLyS) levels were persistently elevated in patients who subsequently relapsed. In conclusion, pathogenesis of HCV-MC vasculitis is mediated by abnormal proliferation of B cells, driven by BLyS, leading to significant effects on non-B cells in mediating symptomatology. Future therapeutics using a combination approach of B cell depletion and proliferation may be desired to achieve long-term remission.http://europepmc.org/articles/PMC5724854?pdf=render |
collection |
DOAJ |
language |
English |
format |
Article |
sources |
DOAJ |
author |
Emily Comstock Cheol-Woo Kim Alison Murphy Benjamin Emmanuel Xi Zhang Michael Sneller Bhawna Poonia Shyamasundaran Kottilil |
spellingShingle |
Emily Comstock Cheol-Woo Kim Alison Murphy Benjamin Emmanuel Xi Zhang Michael Sneller Bhawna Poonia Shyamasundaran Kottilil Transcriptional profiling of PBMCs unravels B cell mediated immunopathogenic imprints of HCV vasculitis. PLoS ONE |
author_facet |
Emily Comstock Cheol-Woo Kim Alison Murphy Benjamin Emmanuel Xi Zhang Michael Sneller Bhawna Poonia Shyamasundaran Kottilil |
author_sort |
Emily Comstock |
title |
Transcriptional profiling of PBMCs unravels B cell mediated immunopathogenic imprints of HCV vasculitis. |
title_short |
Transcriptional profiling of PBMCs unravels B cell mediated immunopathogenic imprints of HCV vasculitis. |
title_full |
Transcriptional profiling of PBMCs unravels B cell mediated immunopathogenic imprints of HCV vasculitis. |
title_fullStr |
Transcriptional profiling of PBMCs unravels B cell mediated immunopathogenic imprints of HCV vasculitis. |
title_full_unstemmed |
Transcriptional profiling of PBMCs unravels B cell mediated immunopathogenic imprints of HCV vasculitis. |
title_sort |
transcriptional profiling of pbmcs unravels b cell mediated immunopathogenic imprints of hcv vasculitis. |
publisher |
Public Library of Science (PLoS) |
series |
PLoS ONE |
issn |
1932-6203 |
publishDate |
2017-01-01 |
description |
B cell depletion therapy using rituximab has been shown to be effective in achieving remission in patients with HCV-mixed cryoglobulinemic (MC) vasculitis. Previously, we have demonstrated abnormalities in peripheral immune cells involving neutrophils, chemotaxis, and innate immune activation among patients with HCV-MC vasculitis when compared to HCV patients without vasculitis. In this study, we evaluated the effect of B cell depletion therapy on transcriptional profiles of peripheral blood mononuclear cells before and after riruximab therapy, in order to unravel the pathogenic mechanism involved in HCV-MC vasculitis induced by abnormal B cell proliferation. DNA microarray analysis was performed using RNA from PBMCs from seven patients with HCV-MC vasculitis and seven normal volunteers. DNA was hybridized to Affymetrix U133A chips. After normalization, differentially expressed gene list with treatment was generated using partitional clustering. RT-PCR, flow cytometry, and enzyme immunoassay (EIA) was used to validate DNA microarray findings. Differentially expressed genes included B cells and non-B cell genes. Validation of genes using purified cell subsets demonstrated distinct effect of B cell depletion therapy on non-B cells, such as monocytes, T cells, and NK cells. Notably, B lymphocyte stimulator (BLyS) levels were persistently elevated in patients who subsequently relapsed. In conclusion, pathogenesis of HCV-MC vasculitis is mediated by abnormal proliferation of B cells, driven by BLyS, leading to significant effects on non-B cells in mediating symptomatology. Future therapeutics using a combination approach of B cell depletion and proliferation may be desired to achieve long-term remission. |
url |
http://europepmc.org/articles/PMC5724854?pdf=render |
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