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Characterization of the Leukocyte Response in Acute Vocal Fold Injury.

King SN, Guille J, Thibeault SL - PLoS ONE (2015)

Bottom Line: Surgical injuries, SWC9+/SWC3- cells exhibited hi CD163+ (p<0.05) at 3-days along with upregulation in TNFα and TGFβ1 mRNA compared to 23-days (p<0.05).Higher levels of IL-10 mRNA were found 1-day post-LPS compared to 5-days (p<0.05).Surgical injury elicited a complex phenotype with early TNFα mRNA and CD163+ and persistent TGFβ1 transcript expression.

View Article: PubMed Central - PubMed

Affiliation: Department of Neurological Surgery, Kentucky Spinal Cord Injury Research Center, University of Louisville, Louisville, Kentucky, United States of America.

ABSTRACT
Macrophages location in the superficial layer of the vocal fold (VF) is not only at the first line of defense, but in a place of physiologic importance to voice quality. This study characterizes and compares macrophage function in two models of acute injury. Porcine VF injuries were created bilaterally by either surgical biopsy or lipopolysaccharide (LPS) (1.5 μg/kg) injection. Animals were sacrificed at 1- or 5-day post LPS or 3-, 7-, or 23-days post-surgical injury (n = 3/time/injury). Flow cytometry characterized immunophenotypes and RT-PCR quantified cytokine gene expression. Uninjured VF were used as controls. Post-surgical and LPS injury, SWC9+/SWC3- cells identified as hi SLA-DR+ (p<0.05) compared to controls along with hi CD16+ expression at 1-day and 3-days respectively compared to all other time points (p<0.05). Surgical injuries, SWC9+/SWC3- cells exhibited hi CD163+ (p<0.05) at 3-days along with upregulation in TNFα and TGFβ1 mRNA compared to 23-days (p<0.05). No measurable changes to IL-12, IFNγ, IL-10, IL-4 mRNA post-surgery. LPS injuries induced upregulation of TNFα, IL-12, IFNγ, IL-10, and IL-4 mRNA at 1- and 5-days compared to controls (p<0.05). Higher levels of IL-10 mRNA were found 1-day post-LPS compared to 5-days (p<0.05). No changes to CD163 or CD80/86 post-LPS were measured. Acute VF injuries revealed a paradigm of markers that appear to associate with each injury. LPS induced a regulatory phenotype indicated by prominent IL-10 mRNA expression. Surgical injury elicited a complex phenotype with early TNFα mRNA and CD163+ and persistent TGFβ1 transcript expression.

No MeSH data available.


Related in: MedlinePlus

Comparison of gene expression levels following LPS (A) or surgical (B) injury using RT-PCR.Standard curves were derived to quantify gene expression and concentrations were normalized to the HPRT–1 housekeeping gene. Data is shown as the mean expression. A statistical significance of p<0.05 is represented by * when compared to normal control and τ when compared to other time points.
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pone.0139260.g006: Comparison of gene expression levels following LPS (A) or surgical (B) injury using RT-PCR.Standard curves were derived to quantify gene expression and concentrations were normalized to the HPRT–1 housekeeping gene. Data is shown as the mean expression. A statistical significance of p<0.05 is represented by * when compared to normal control and τ when compared to other time points.

Mentions: To determine inflammatory profiles after injury, gene expression of TNFα, IL–10, IL–12, IL–4, TGFβ1, and IFNγ was measured using RT-PCR from RNA procured from the vocal fold cover. Following LPS injection, expression of TNFα (p<0.0001), IL–10 (p<0.0001), IL–4 (p<0.0001), IFNγ (p<0.01), and IL–12 (p<0.0001) mRNA was significantly higher after 1- and 5-days compared to uninjured controls (Fig 6A). Statistical differences in IL–10 and IFNγ mRNA expression were found between time points. IL–10 gene expression levels peaked at 3-days compared to 5-days post LPS (p<0.001), where IFNγ expression significantly increased at later time point compared to 3-days (p<0.05). No changes between time points for TNFα, IL–4, or IL–12 were measured. Following surgical trauma, differences in TNFα and TGFβ1 expression were appreciated (Fig 6B). TNFα mRNA expression was significantly highest after 3-days in comparison to all other time points (p< 0.001) and non-injured control (p< 0.01). TGFβ1 expression peaked at 3-days (p<0.01) and 7-days (p< 0.05) post surgery with significant decline after 23-days. Significant increases also noted between 3-days post surgery and non-injured controls (p< 0.01). No changes in IL–10, IL–4, IFNγ, or IL–12 gene expression were found post-surgery.


Characterization of the Leukocyte Response in Acute Vocal Fold Injury.

King SN, Guille J, Thibeault SL - PLoS ONE (2015)

Comparison of gene expression levels following LPS (A) or surgical (B) injury using RT-PCR.Standard curves were derived to quantify gene expression and concentrations were normalized to the HPRT–1 housekeeping gene. Data is shown as the mean expression. A statistical significance of p<0.05 is represented by * when compared to normal control and τ when compared to other time points.
© Copyright Policy
Related In: Results  -  Collection

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getmorefigures.php?uid=PMC4591973&req=5

pone.0139260.g006: Comparison of gene expression levels following LPS (A) or surgical (B) injury using RT-PCR.Standard curves were derived to quantify gene expression and concentrations were normalized to the HPRT–1 housekeeping gene. Data is shown as the mean expression. A statistical significance of p<0.05 is represented by * when compared to normal control and τ when compared to other time points.
Mentions: To determine inflammatory profiles after injury, gene expression of TNFα, IL–10, IL–12, IL–4, TGFβ1, and IFNγ was measured using RT-PCR from RNA procured from the vocal fold cover. Following LPS injection, expression of TNFα (p<0.0001), IL–10 (p<0.0001), IL–4 (p<0.0001), IFNγ (p<0.01), and IL–12 (p<0.0001) mRNA was significantly higher after 1- and 5-days compared to uninjured controls (Fig 6A). Statistical differences in IL–10 and IFNγ mRNA expression were found between time points. IL–10 gene expression levels peaked at 3-days compared to 5-days post LPS (p<0.001), where IFNγ expression significantly increased at later time point compared to 3-days (p<0.05). No changes between time points for TNFα, IL–4, or IL–12 were measured. Following surgical trauma, differences in TNFα and TGFβ1 expression were appreciated (Fig 6B). TNFα mRNA expression was significantly highest after 3-days in comparison to all other time points (p< 0.001) and non-injured control (p< 0.01). TGFβ1 expression peaked at 3-days (p<0.01) and 7-days (p< 0.05) post surgery with significant decline after 23-days. Significant increases also noted between 3-days post surgery and non-injured controls (p< 0.01). No changes in IL–10, IL–4, IFNγ, or IL–12 gene expression were found post-surgery.

Bottom Line: Surgical injuries, SWC9+/SWC3- cells exhibited hi CD163+ (p<0.05) at 3-days along with upregulation in TNFα and TGFβ1 mRNA compared to 23-days (p<0.05).Higher levels of IL-10 mRNA were found 1-day post-LPS compared to 5-days (p<0.05).Surgical injury elicited a complex phenotype with early TNFα mRNA and CD163+ and persistent TGFβ1 transcript expression.

View Article: PubMed Central - PubMed

Affiliation: Department of Neurological Surgery, Kentucky Spinal Cord Injury Research Center, University of Louisville, Louisville, Kentucky, United States of America.

ABSTRACT
Macrophages location in the superficial layer of the vocal fold (VF) is not only at the first line of defense, but in a place of physiologic importance to voice quality. This study characterizes and compares macrophage function in two models of acute injury. Porcine VF injuries were created bilaterally by either surgical biopsy or lipopolysaccharide (LPS) (1.5 μg/kg) injection. Animals were sacrificed at 1- or 5-day post LPS or 3-, 7-, or 23-days post-surgical injury (n = 3/time/injury). Flow cytometry characterized immunophenotypes and RT-PCR quantified cytokine gene expression. Uninjured VF were used as controls. Post-surgical and LPS injury, SWC9+/SWC3- cells identified as hi SLA-DR+ (p<0.05) compared to controls along with hi CD16+ expression at 1-day and 3-days respectively compared to all other time points (p<0.05). Surgical injuries, SWC9+/SWC3- cells exhibited hi CD163+ (p<0.05) at 3-days along with upregulation in TNFα and TGFβ1 mRNA compared to 23-days (p<0.05). No measurable changes to IL-12, IFNγ, IL-10, IL-4 mRNA post-surgery. LPS injuries induced upregulation of TNFα, IL-12, IFNγ, IL-10, and IL-4 mRNA at 1- and 5-days compared to controls (p<0.05). Higher levels of IL-10 mRNA were found 1-day post-LPS compared to 5-days (p<0.05). No changes to CD163 or CD80/86 post-LPS were measured. Acute VF injuries revealed a paradigm of markers that appear to associate with each injury. LPS induced a regulatory phenotype indicated by prominent IL-10 mRNA expression. Surgical injury elicited a complex phenotype with early TNFα mRNA and CD163+ and persistent TGFβ1 transcript expression.

No MeSH data available.


Related in: MedlinePlus