Baseline demographics, pre-existing health status factors and study outcomes for the burn cohort vs. uninjured cohort.

<p>Baseline demographics, pre-existing health status factors and study outcomes for the burn cohort vs. uninjured cohort.</p

Reduced CD8 and increased NK/NKT response to viral infection in the airway draining lymph nodes after burn injury.

<p>At day 4 and day 10 post-infection no significant difference was observed in total cell number of CD3, CD4 and CD8 cell numbers (a-d). CD4 proliferation (Ki67+), antigen experience (CD44+), activation (CD25+) and CD4 Treg frequency were all not significantly different between burn and sham treatment groups (e-h). CD8 proliferation appeared to be reduced after burn injury (p = 0.056, i) with a loss of the significant increase in CD8 cells during infection compared to naïve mice seen in sham injured animals (l), but no other changes in the CD8 profile were observed (j-k). NK and NKT cell number appear to be increased at day 4 and day 10 post-infection respectively in the burn injury group (m, n, p = 0.091, p = 0.061). Gran B positive NK and NKT cells were both significantly elevated at day 4 post-infection in the burn injury group (o, p). * p<0.05.</p

Adjusted incident rate ratios (IRR) and 95% confidence intervals (CI) for recurrent admissions for respiratory sub-conditions for sub cohorts defined by TBSA burn severity versus uninjured cohort.

<p>Adjusted incident rate ratios (IRR) and 95% confidence intervals (CI) for recurrent admissions for respiratory sub-conditions for sub cohorts defined by TBSA burn severity versus uninjured cohort.</p

Keratinocytes on RADA16-Collagen I (1% wt)

Live cell tracking migration assay with keratinocytes on RADA16-Collagen I (1% wt)

Keratinocytes on RADA16-Fibronectin (1% wt)

Live cell tracking migration assay with keratinocytes on RADA16-Fibronectin (1% wt)

Keratinocytes on RADA16 (1% wt)

Live cell tracking migration assay with keratinocytes on RADA16 (1% wt)

Fibroblasts on RADA16-Fibronectin (1% wt)

Live cell tracking migration assay with fibroblasts on RADA16-Fibronectin (1% wt)

Fibroblasts on RADA16-Collagen I (1% wt)

Live cell tracking migration assay with fibroblasts on RADA16-Collagen I (1% wt)

Data_Sheet_1_Non-severe thermal burn injuries induce long-lasting downregulation of gene expression in cortical excitatory neurons and microglia.docx

Burn injuries are devastating traumas, often leading to life-long consequences that extend beyond the observable burn scar. In the context of the nervous system, burn injury patients commonly develop chronic neurological disorders and have been suggested to have impaired motor cortex function, but the long-lasting impact on neurons and glia in the brain is unknown. Using a mouse model of non-severe burn injury, excitatory and inhibitory neurons in the primary motor cortex were labelled with fluorescent proteins using adeno-associated viruses (AAVs). A total of 5 weeks following the burn injury, virus labelled excitatory and inhibitory neurons were isolated using fluorescence-activated cell sorting (FACS). In addition, microglia and astrocytes from the remaining cortical tissue caudal to the motor cortex were immunolabelled and isolated with FACS. Whole transcriptome RNA-sequencing was used to identify any long-lasting changes to gene expression in the different cell types. RNA-seq analysis showed changes to the expression of a small number of genes with known functions in excitatory neurons and microglia, but not in inhibitory neurons or astrocytes. Specifically, genes related to GABA-A receptors in excitatory neurons and several cellular functions in microglia were found to be downregulated in burn injured mice. These findings suggest that non-severe burn injuries lead to long lasting transcriptomic changes in the brain, but only in specific cell types. Our findings provide a broad overview of the long-lasting impact of burn injuries on the central nervous system which may help identify potential therapeutic targets to prevent neurological dysfunction in burn patients.</p

Table_1_Non-severe thermal burn injuries induce long-lasting downregulation of gene expression in cortical excitatory neurons and microglia.PDF

Burn injuries are devastating traumas, often leading to life-long consequences that extend beyond the observable burn scar. In the context of the nervous system, burn injury patients commonly develop chronic neurological disorders and have been suggested to have impaired motor cortex function, but the long-lasting impact on neurons and glia in the brain is unknown. Using a mouse model of non-severe burn injury, excitatory and inhibitory neurons in the primary motor cortex were labelled with fluorescent proteins using adeno-associated viruses (AAVs). A total of 5 weeks following the burn injury, virus labelled excitatory and inhibitory neurons were isolated using fluorescence-activated cell sorting (FACS). In addition, microglia and astrocytes from the remaining cortical tissue caudal to the motor cortex were immunolabelled and isolated with FACS. Whole transcriptome RNA-sequencing was used to identify any long-lasting changes to gene expression in the different cell types. RNA-seq analysis showed changes to the expression of a small number of genes with known functions in excitatory neurons and microglia, but not in inhibitory neurons or astrocytes. Specifically, genes related to GABA-A receptors in excitatory neurons and several cellular functions in microglia were found to be downregulated in burn injured mice. These findings suggest that non-severe burn injuries lead to long lasting transcriptomic changes in the brain, but only in specific cell types. Our findings provide a broad overview of the long-lasting impact of burn injuries on the central nervous system which may help identify potential therapeutic targets to prevent neurological dysfunction in burn patients.</p