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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 SWC9+/SWC3- cells and CD16+/SWC9- cells in the vocal fold lamina propria following LPS (A & B) or surgical (C & D) injury (d = days).Cells were stained for SWC9, SWC3, and CD16. * represents a statistical significance of p<0.05 when compared to other time points.
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pone.0139260.g003: Comparison of SWC9+/SWC3- cells and CD16+/SWC9- cells in the vocal fold lamina propria following LPS (A & B) or surgical (C & D) injury (d = days).Cells were stained for SWC9, SWC3, and CD16. * represents a statistical significance of p<0.05 when compared to other time points.

Mentions: To determine the immune cell response in the vocal fold following injury, we performed seven-color flow cytometry on cells isolated from the vocal folds using SWC9, SWC3, CD16, CD163, CD80/86, SLA-DR, and dead cell staining. After gating for live and single cells, SWC9+/SWC3- cell and CD16+/SWC9- cell populations were then identified in vocal fold. LPS injury resulted in inverse, re-distribution of macrophage and neutrophil like cells in the tissue (Fig 3A & 3B). The percentage of cells expressing SWC3-/SWC9+ significantly decreased after 1-day in comparison to all other time points (0-day p<0.001; 5-day p< 0.01), while percentage of cells expressing CD16+/SWC9- significantly increased in comparison (0-day and 5-day p< 0.05). In contrast, SWC9+/SWC3- cells concentrations following surgical injury remained practically unchanged from 3 to 23 days (Fig 3C & 3D). However, the percent of CD16+/SWC9- cells was significantly increased at day 3 in comparison to all other time points (p< 0.05).


Characterization of the Leukocyte Response in Acute Vocal Fold Injury.

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

Comparison of SWC9+/SWC3- cells and CD16+/SWC9- cells in the vocal fold lamina propria following LPS (A & B) or surgical (C & D) injury (d = days).Cells were stained for SWC9, SWC3, and CD16. * represents a statistical significance of p<0.05 when compared to other time points.
© Copyright Policy
Related In: Results  -  Collection

License
Show All Figures
getmorefigures.php?uid=PMC4591973&req=5

pone.0139260.g003: Comparison of SWC9+/SWC3- cells and CD16+/SWC9- cells in the vocal fold lamina propria following LPS (A & B) or surgical (C & D) injury (d = days).Cells were stained for SWC9, SWC3, and CD16. * represents a statistical significance of p<0.05 when compared to other time points.
Mentions: To determine the immune cell response in the vocal fold following injury, we performed seven-color flow cytometry on cells isolated from the vocal folds using SWC9, SWC3, CD16, CD163, CD80/86, SLA-DR, and dead cell staining. After gating for live and single cells, SWC9+/SWC3- cell and CD16+/SWC9- cell populations were then identified in vocal fold. LPS injury resulted in inverse, re-distribution of macrophage and neutrophil like cells in the tissue (Fig 3A & 3B). The percentage of cells expressing SWC3-/SWC9+ significantly decreased after 1-day in comparison to all other time points (0-day p<0.001; 5-day p< 0.01), while percentage of cells expressing CD16+/SWC9- significantly increased in comparison (0-day and 5-day p< 0.05). In contrast, SWC9+/SWC3- cells concentrations following surgical injury remained practically unchanged from 3 to 23 days (Fig 3C & 3D). However, the percent of CD16+/SWC9- cells was significantly increased at day 3 in comparison to all other time points (p< 0.05).

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