hbd-1 elisa developing kits  (PeproTech)

Journal: PLoS ONE

Article Title: Lineage − CD34 + CD31 + Cells That Appear in Association with Severe Burn Injury Are Inhibitory on the Production of Antimicrobial Peptides by Epidermal Keratinocytes

doi: 10.1371/journal.pone.0082926

Figure Lengend Snippet: HBD-1 production. Lineage − CD34 + cells (1×10 5 cells/ml, upper chamber), isolated from 5 healthy donors (#1∼#5) and 20 burn patients (#1∼#20), were transwell-cultured with NHEK (1×10 5 cells/ml, lower chamber) for 36 hours. After removal of the upper chamber, NHEK in the lower chamber were cultured for an additional 36 hours. Culture fluids obtained were assayed for HBD-1 by ELISA. shows independent experiments performed using blood specimens from 5 healthy donors and 20 burn patients, and shows mean ± SEM of the results shown in . * P <0.001 vs control. B and C. HBD-1 mRNA expression. Lineage − CD34 + cells (1×10 5 cells/ml, upper chamber), isolated from peripheral blood of 5 healthy donors (#1∼#5, B ) and 5 severely burn patients (#1, #16, #17, #18, #19, C ), were transwell-cultured with NHEK (1×10 5 cells/ml, lower chamber) for 36 hours. After removal of the upper chamber, NHEK in the lower chamber were analyzed for HBD-1 mRNA by RT-PCR.

Article Snippet: Human β-defensin-1 (HBD-1) kits were purchased from PeproTech (Rocky Hill, NJ, USA).

Techniques: Isolation, Cell Culture, Enzyme-linked Immunosorbent Assay, Expressing, Reverse Transcription Polymerase Chain Reaction

Journal: PLoS ONE

Article Title: Lineage − CD34 + CD31 + Cells That Appear in Association with Severe Burn Injury Are Inhibitory on the Production of Antimicrobial Peptides by Epidermal Keratinocytes

doi: 10.1371/journal.pone.0082926

Figure Lengend Snippet: Inhibition of HBD production in NHEK cultures supplemented with culture fluids of lineage − CD34 + CD31 + cells. Culture fluids were harvested 48 hours after cultivation of lineage − CD34 + CD31 + cells (1×10 6 cells/ml) derived from burn patients #8, #9, #13, #14 and #15 (open circles). The culture fluids of lineage − CD34 + CD31 − cells of healthy donors #1∼#5 (filled circles) were utilized as a control. Five to 40% (v/v) of these culture fluids were added to cultures of NHEK (1×10 5 cells/ml). Culture fluids harvested from NHEK cultures were assayed for HBD-1 by ELISA. * P <0.01 vs control. B–D. Production of IL-10, IL-13 and CCL2 by lineage − CD34 + CD31 + cells. Lineage − CD34 + CD31 + cells (1×10 5 cells/ml, open circles) isolated from peripheral blood of burn patients #8, #9, #13, #14 and #15 were cultured for 12 to 48 hours. As controls, peripheral blood lineage − CD34 + CD31 − cells of healthy donors #1∼#5 (filled circles) were cultured under the same conditions. Supernatants obtained were assayed for IL-10 ( B ), IL-13 ( C ) and CCL2 ( D ) by ELISA. * P <0.05, ** P <0.01 vs lineage − CD34 + CD31 − cells.

Article Snippet: Human β-defensin-1 (HBD-1) kits were purchased from PeproTech (Rocky Hill, NJ, USA).

Techniques: Inhibition, Derivative Assay, Enzyme-linked Immunosorbent Assay, Isolation, Cell Culture

Journal: PLoS ONE

Article Title: Lineage − CD34 + CD31 + Cells That Appear in Association with Severe Burn Injury Are Inhibitory on the Production of Antimicrobial Peptides by Epidermal Keratinocytes

doi: 10.1371/journal.pone.0082926

Figure Lengend Snippet: Effect of mAbs against IL-10, CCL2 and IL-13 on the suppressor activities of lineage − CD34 + CD31 + cell-culture fluids on HBD production by NHEK. NHEK (1×10 5 cells/ml) were cultured with fresh media supplemented with 20% (v/v) of the culture fluids that were previously treated with mAb for IL-10, IL-13 and CCL2 (2.5 µg/ml each) or a combination of these mAbs. Forty-eight hours after cultivation, culture fluids harvested were assayed for HBD-1. displays one of the representative results shown in independent experiments using culture fluids of 5 different lineage − CD34 + CD31 + cell preparations derived from burn patients #8, #9, #13, #14 and #15. B. Effect of rIL-10 and rCCL2 on HBD-1 production by NHEK. NHEK (1×10 5 cells/ml) were treated with rIL-10 (1 ng/ml), rCCL2 (10 ng/ml) or a combination of rIL-10 and rCCL2. Culture fluids obtained 24 hours after treatment were assayed for HBD-1. * P <0.01, ** P <0.001 vs NHEK cultured with media.

Article Snippet: Human β-defensin-1 (HBD-1) kits were purchased from PeproTech (Rocky Hill, NJ, USA).

Techniques: Cell Culture, Derivative Assay