Journal: The Journal of trauma

Article Title: Th17 (IFN γ − IL17 + ) CD4 + T Cells Generated After Burn Injury May Be a Novel Cellular Mechanism for Postburn Immunosuppression

doi: 10.1097/TA.0b013e31820d18a6

Figure Lengend Snippet: Wound-draining lymph nodes, but not the spleen, contain Th17 cells 3 days and 14 days after burn injury. Cells were harvested from spleens, as well as inguinal and axillary PLNs, 3 days or 14 days after a 20% TBSA, full-thickness burn, or sham injury. Bulk PLN cells were stimulated in vitro and intracellular cytokine staining was performed, and flow cytometry was used to measure IL-17 and IFN-γ production by CD3+ CD4+ cells. Representative examples of the flow cytometry histograms for each lymphoid organ are provided.

Article Snippet: Th17 In Vitro Cell Polarization Purified CD4 + T cells from burn and sham mice (1 × 10 6 cells/mL) cells were stimulated with plate bound antimouse CD3 antibody (1 μ g/well; Becton Dickinson, San Diego, CA) and soluble antimouse CD28 antibody (5 μ g/mL; BD Pharmingen, San Diego, CA) in the presence of IL-6 (50 ng/mL; BD Pharmingen, San Diego, CA) and TGF- β (1 ng/mL; PeproTech, Rocky Hill, NJ) cytokines for a total of 4 days in 0.5 mL of complete RPMI (10% fetal calf serum) in 48-well flat-bottom plates.

Techniques: In Vitro, Staining, Flow Cytometry

Journal: The Journal of trauma

Article Title: Th17 (IFN γ − IL17 + ) CD4 + T Cells Generated After Burn Injury May Be a Novel Cellular Mechanism for Postburn Immunosuppression

doi: 10.1097/TA.0b013e31820d18a6

Figure Lengend Snippet: Percentage of CD4+ T cells that produce IL-17 are significantly higher in burn compared with sham. Cells were harvested from spleens, as well as inguinal and axillary PLNs, 3 days or 14 days after a 20% TBSA, full-thickness burn, or sham injury. Bulk PLN cells were stimulated in vitro and intracellular cytokine staining was performed, and flow cytometry was used to measure IL-17 and IFN-γ production by CD3+ CD4+ T cells. Data were expressed as mean ± standard error of the mean, *p ≤ 0.05; **p ≤ 0.005 compared with matched sham controls by Student’s t test (n = 4–6 mice/group).

Article Snippet: Th17 In Vitro Cell Polarization Purified CD4 + T cells from burn and sham mice (1 × 10 6 cells/mL) cells were stimulated with plate bound antimouse CD3 antibody (1 μ g/well; Becton Dickinson, San Diego, CA) and soluble antimouse CD28 antibody (5 μ g/mL; BD Pharmingen, San Diego, CA) in the presence of IL-6 (50 ng/mL; BD Pharmingen, San Diego, CA) and TGF- β (1 ng/mL; PeproTech, Rocky Hill, NJ) cytokines for a total of 4 days in 0.5 mL of complete RPMI (10% fetal calf serum) in 48-well flat-bottom plates.

Techniques: In Vitro, Staining, Flow Cytometry

Journal: The Journal of trauma

Article Title: Th17 (IFN γ − IL17 + ) CD4 + T Cells Generated After Burn Injury May Be a Novel Cellular Mechanism for Postburn Immunosuppression

doi: 10.1097/TA.0b013e31820d18a6

Figure Lengend Snippet: CD4+ T cells from burn and sham mice have similar abilities to polarize to Th17 cells. Cells were harvested from various PLNs at 3 days or 14 days after a 20% TBSA, full-thickness burn, or sham injury. CD4+ T cells were enriched by negative magnetic selection and cultured with plate bound anti-CD3, soluble anti-CD28, IL-6, and TGF-β for 4 days. IL-17 and IFN-γ production by CD3+ CD4+ cells was analyzed using flow cytometric staining. Representative examples of the histograms for each timepoint are provided.

Article Snippet: Th17 In Vitro Cell Polarization Purified CD4 + T cells from burn and sham mice (1 × 10 6 cells/mL) cells were stimulated with plate bound antimouse CD3 antibody (1 μ g/well; Becton Dickinson, San Diego, CA) and soluble antimouse CD28 antibody (5 μ g/mL; BD Pharmingen, San Diego, CA) in the presence of IL-6 (50 ng/mL; BD Pharmingen, San Diego, CA) and TGF- β (1 ng/mL; PeproTech, Rocky Hill, NJ) cytokines for a total of 4 days in 0.5 mL of complete RPMI (10% fetal calf serum) in 48-well flat-bottom plates.

Techniques: Selection, Cell Culture, Staining

Journal: The Journal of trauma

Article Title: Th17 (IFN γ − IL17 + ) CD4 + T Cells Generated After Burn Injury May Be a Novel Cellular Mechanism for Postburn Immunosuppression

doi: 10.1097/TA.0b013e31820d18a6

Figure Lengend Snippet: Burn serum does not contain a soluble factor that increases that ability of CD4+ T cells to polarize to a Th17 phenotype. Cells were harvested from various PLNs at 14 days after a 20% TBSA, full-thickness burn, or sham injury. Burn serum was also collected at 14 days after burn injury. CD4+ T cells were enriched by negative magnetic selection. Cells were then cultured with plate bound anti-CD3, soluble anti-CD28, IL-6, and TGF-β in the presence or absence of burn serum for 4 days. IL-17 and IFN-γ production by CD3+ CD4+ cells was analyzed using flow cytometric staining. Isotype refers to staining with isotype control antibodies. Data were expressed as mean ± standard error of the mean (n = 4–6 mice/group).

Article Snippet: Th17 In Vitro Cell Polarization Purified CD4 + T cells from burn and sham mice (1 × 10 6 cells/mL) cells were stimulated with plate bound antimouse CD3 antibody (1 μ g/well; Becton Dickinson, San Diego, CA) and soluble antimouse CD28 antibody (5 μ g/mL; BD Pharmingen, San Diego, CA) in the presence of IL-6 (50 ng/mL; BD Pharmingen, San Diego, CA) and TGF- β (1 ng/mL; PeproTech, Rocky Hill, NJ) cytokines for a total of 4 days in 0.5 mL of complete RPMI (10% fetal calf serum) in 48-well flat-bottom plates.

Techniques: Selection, Cell Culture, Staining, Control