Toll-like receptor chaperone HSP90B1 and the immune response to Mycobacteria.
<h4>Rationale</h4>HSP90B1, also known as gp96, is a chaperone for multiple Toll-like receptors (TLRs) and is necessary for TLR-mediated inflammatory responses in murine myeloid cells. The molecule is also expressed in T-cells though its specific role is unknown. We hypothesized that huma...
| Main Authors: | , , , , , , , , , , , , , |
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| Format: | Article |
| Language: | English |
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Public Library of Science (PLoS)
2018-01-01
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| Series: | PLoS ONE |
| Online Access: | https://doi.org/10.1371/journal.pone.0208940 |
| _version_ | 1818681541904039936 |
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| author | Andrew D Graustein Elizabeth A Misch Munyaradzi Musvosvi Muki Shey Javeed A Shah Chetan Seshadri Augustine Ajuogu Kathryn Bowman Humphrey Mulenga Ashley Veldsman Willem A Hanekom Mark Hatherill Thomas J Scriba Thomas R Hawn |
| author_facet | Andrew D Graustein Elizabeth A Misch Munyaradzi Musvosvi Muki Shey Javeed A Shah Chetan Seshadri Augustine Ajuogu Kathryn Bowman Humphrey Mulenga Ashley Veldsman Willem A Hanekom Mark Hatherill Thomas J Scriba Thomas R Hawn |
| author_sort | Andrew D Graustein |
| collection | DOAJ |
| description | <h4>Rationale</h4>HSP90B1, also known as gp96, is a chaperone for multiple Toll-like receptors (TLRs) and is necessary for TLR-mediated inflammatory responses in murine myeloid cells. The molecule is also expressed in T-cells though its specific role is unknown. We hypothesized that human HSP90B1 regulates monocyte and T-cell responses to Mycobacterium tuberculosis (Mtb) and bacilli Calmette-Guerin (BCG) and that its variants are associated with susceptibility to TB disease.<h4>Methods</h4>We screened 17 haplotype-tagging SNPs in the HSP90B1 gene region for association with BCG-induced T-cell cytokine responses using both an ex-vivo whole blood assay (N = 295) and an intracellular cytokine staining assay (N = 180) on samples collected 10 weeks after birth. Using a case-control study design, we evaluated the same SNPs for association with TB disease in a South African pediatric cohort (N = 217 cases, 604 controls). A subset of these SNPs was evaluated for association with HSP90B1 expression in human monocytes, monocyte-derived dendritic cells, and T-cells using RT-PCR. Lastly, we used CRISPR/Cas9 gene editing to knock down HSP90B1 expression in a human monocyte cell line (U937). Knockdown and control cell lines were tested for TLR surface expression and control of Mtb replication.<h4>Results</h4>We identified three SNPs, rs10507172, rs10507173 and rs1920413, that were associated with BCG-induced IL-2 secretion (p = 0.017 for rs10507172 and p = 0.03 for rs10507173 and rs1920413, Mann-Whitney, dominant model). SNPs rs10507172 and rs10507173 were associated with TB disease in an unadjusted analysis (p = 0.036 and 0.025, respectively, dominant model) that strengthened with sensitivity analysis of the definite TB cases, which included only those patients with microbiologically confirmed Mtb (p = 0.007 and 0.012, respectively). Knockdowns of HSP90B1 in monocyte cell lines with CRISPR did not alter TLR2 surface expression nor influence Mtb replication relative to controls.<h4>Conclusion</h4>Among infants, an HSP90B1 gene-region variant is associated with BCG-induced IL-2 production and may be associated with protection from TB disease. HSP90B1 knockdown in human monocyte-like cell lines did not influence TLR2 surface localization nor Mtb replication. Together, these data suggest that HSP90B1 regulates T-cell, but not monocyte, responses to mycobacteria in humans. |
| first_indexed | 2024-12-17T10:04:36Z |
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| id | doaj.art-642f66a203454435be42a7e9ba539e90 |
| institution | Directory Open Access Journal |
| issn | 1932-6203 |
| language | English |
| last_indexed | 2024-12-17T10:04:36Z |
| publishDate | 2018-01-01 |
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| spelling | doaj.art-642f66a203454435be42a7e9ba539e902022-12-21T21:53:11ZengPublic Library of Science (PLoS)PLoS ONE1932-62032018-01-011312e020894010.1371/journal.pone.0208940Toll-like receptor chaperone HSP90B1 and the immune response to Mycobacteria.Andrew D GrausteinElizabeth A MischMunyaradzi MusvosviMuki SheyJaveed A ShahChetan SeshadriAugustine AjuoguKathryn BowmanHumphrey MulengaAshley VeldsmanWillem A HanekomMark HatherillThomas J ScribaThomas R Hawn<h4>Rationale</h4>HSP90B1, also known as gp96, is a chaperone for multiple Toll-like receptors (TLRs) and is necessary for TLR-mediated inflammatory responses in murine myeloid cells. The molecule is also expressed in T-cells though its specific role is unknown. We hypothesized that human HSP90B1 regulates monocyte and T-cell responses to Mycobacterium tuberculosis (Mtb) and bacilli Calmette-Guerin (BCG) and that its variants are associated with susceptibility to TB disease.<h4>Methods</h4>We screened 17 haplotype-tagging SNPs in the HSP90B1 gene region for association with BCG-induced T-cell cytokine responses using both an ex-vivo whole blood assay (N = 295) and an intracellular cytokine staining assay (N = 180) on samples collected 10 weeks after birth. Using a case-control study design, we evaluated the same SNPs for association with TB disease in a South African pediatric cohort (N = 217 cases, 604 controls). A subset of these SNPs was evaluated for association with HSP90B1 expression in human monocytes, monocyte-derived dendritic cells, and T-cells using RT-PCR. Lastly, we used CRISPR/Cas9 gene editing to knock down HSP90B1 expression in a human monocyte cell line (U937). Knockdown and control cell lines were tested for TLR surface expression and control of Mtb replication.<h4>Results</h4>We identified three SNPs, rs10507172, rs10507173 and rs1920413, that were associated with BCG-induced IL-2 secretion (p = 0.017 for rs10507172 and p = 0.03 for rs10507173 and rs1920413, Mann-Whitney, dominant model). SNPs rs10507172 and rs10507173 were associated with TB disease in an unadjusted analysis (p = 0.036 and 0.025, respectively, dominant model) that strengthened with sensitivity analysis of the definite TB cases, which included only those patients with microbiologically confirmed Mtb (p = 0.007 and 0.012, respectively). Knockdowns of HSP90B1 in monocyte cell lines with CRISPR did not alter TLR2 surface expression nor influence Mtb replication relative to controls.<h4>Conclusion</h4>Among infants, an HSP90B1 gene-region variant is associated with BCG-induced IL-2 production and may be associated with protection from TB disease. HSP90B1 knockdown in human monocyte-like cell lines did not influence TLR2 surface localization nor Mtb replication. Together, these data suggest that HSP90B1 regulates T-cell, but not monocyte, responses to mycobacteria in humans.https://doi.org/10.1371/journal.pone.0208940 |
| spellingShingle | Andrew D Graustein Elizabeth A Misch Munyaradzi Musvosvi Muki Shey Javeed A Shah Chetan Seshadri Augustine Ajuogu Kathryn Bowman Humphrey Mulenga Ashley Veldsman Willem A Hanekom Mark Hatherill Thomas J Scriba Thomas R Hawn Toll-like receptor chaperone HSP90B1 and the immune response to Mycobacteria. PLoS ONE |
| title | Toll-like receptor chaperone HSP90B1 and the immune response to Mycobacteria. |
| title_full | Toll-like receptor chaperone HSP90B1 and the immune response to Mycobacteria. |
| title_fullStr | Toll-like receptor chaperone HSP90B1 and the immune response to Mycobacteria. |
| title_full_unstemmed | Toll-like receptor chaperone HSP90B1 and the immune response to Mycobacteria. |
| title_short | Toll-like receptor chaperone HSP90B1 and the immune response to Mycobacteria. |
| title_sort | toll like receptor chaperone hsp90b1 and the immune response to mycobacteria |
| url | https://doi.org/10.1371/journal.pone.0208940 |
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