Journal: Arthritis Research & Therapy

Article Title: Tumor necrosis factor and norepinephrine lower the levels of human neutrophil peptides 1-3 secretion by mixed synovial tissue cultures in osteoarthritis and rheumatoid arthritis

doi: 10.1186/ar3044

Figure Lengend Snippet: Immunohistochemistry of human neutrophil peptides 1-3 (HNP1-3) and human β-defensin 2 (HBD 2) in the synovial lining and sublining area of an RA patient (similar in OA patients) . Antibodies to HNP1-3 stained positive in the lining and adjacent sublining area, whereas HBD-2 was found only in deep sublining zones. Magnification 400×.

Article Snippet: Then, sections were incubated with either monoclonal mouse anti-human antibodies against HNP1-3 (BMA Biomedicals, Augst, Switzerland, no. T 1034; dilution, 1:1,000; these antibodies recognize HNP-1 to -3) or polyclonal rabbit anti-human antibodies against HBD-2 (Biologo, Kronshagen, Germany, no. DEF002; dilution, 1:100).

Techniques: Immunohistochemistry, Staining

Journal: Arthritis Research & Therapy

Article Title: Tumor necrosis factor and norepinephrine lower the levels of human neutrophil peptides 1-3 secretion by mixed synovial tissue cultures in osteoarthritis and rheumatoid arthritis

doi: 10.1186/ar3044

Figure Lengend Snippet: Double immunofluorescence of human neutrophil peptides 1-3 (HNP1-3) and human β-defensin 2 (HBD 2) in synovial tissue of an RA patient . Arrowheads show double-positive cells. Left panels: HNP1-3 (red staining) were co-localized with neutrophil elastase (green staining), as indicated by the yellow color in this overlay image. Right panels : HBD-2 (red fluorescence) was rarely detected. Some fibroblasts (green fluorescence in the upper right panel) and some CD19 + B lymphocytes (green fluorescence in the lower right panel) stained positive for HBD-2 (yellow overlay). Magnification 400×.

Article Snippet: Then, sections were incubated with either monoclonal mouse anti-human antibodies against HNP1-3 (BMA Biomedicals, Augst, Switzerland, no. T 1034; dilution, 1:1,000; these antibodies recognize HNP-1 to -3) or polyclonal rabbit anti-human antibodies against HBD-2 (Biologo, Kronshagen, Germany, no. DEF002; dilution, 1:100).

Techniques: Immunofluorescence, Staining, Fluorescence

Journal: Arthritis Research & Therapy

Article Title: Tumor necrosis factor and norepinephrine lower the levels of human neutrophil peptides 1-3 secretion by mixed synovial tissue cultures in osteoarthritis and rheumatoid arthritis

doi: 10.1186/ar3044

Figure Lengend Snippet: Secretion of defensins from synovial tissue/cells and density of synovial neutrophils . (a) Levels of human neutrophil peptides 1-3 (HNP1-3) in superfusate of synovial tissue of two patients with rheumatoid arthritis (RA) and osteoarthritis (OA) (four replicates per patient). The dotted line indicates the detection limit. (b) Levels of human β-defensin-2 (HBD 2) in supernatant of synovial tissue of two patients with RA and OA (four replicates per patient). The dotted line indicates the detection limit. (c) Levels of defensins in supernatant of cultured mixed synovial cells over time. Compare the levels with the detection levels in panels (a) and (b). After 20 hours, HNP1-3 were in the detectable range, which was not the case for HBD-2. The data of four OA patients are given (two replicates of every patient), which was similar in RA patients (data not shown). (d) Density of synovial neutrophils in RA and OA synovial tissue. Every symbol represents the average density of neutrophils of one patient, as measured in 17 high-power fields (400×) of two to three synovial tissue sections, including deep sublining areas.

Article Snippet: Then, sections were incubated with either monoclonal mouse anti-human antibodies against HNP1-3 (BMA Biomedicals, Augst, Switzerland, no. T 1034; dilution, 1:1,000; these antibodies recognize HNP-1 to -3) or polyclonal rabbit anti-human antibodies against HBD-2 (Biologo, Kronshagen, Germany, no. DEF002; dilution, 1:100).

Techniques: Cell Culture

Journal: Arthritis Research & Therapy

Article Title: Tumor necrosis factor and norepinephrine lower the levels of human neutrophil peptides 1-3 secretion by mixed synovial tissue cultures in osteoarthritis and rheumatoid arthritis

doi: 10.1186/ar3044

Figure Lengend Snippet: Influence of tumor necrosis factor (TNF) on levels of human neutrophil peptides 1-3 (HNP1-3) of mixed synoviocytes . The data are derived from five OA and five RA patients (two replicates of every patient). Control HNP1-3 levels were 58.3 ± 7.0 pg/ml in OA patients and 147.2 ± 37.5 pg/ml in RA patients.

Article Snippet: Then, sections were incubated with either monoclonal mouse anti-human antibodies against HNP1-3 (BMA Biomedicals, Augst, Switzerland, no. T 1034; dilution, 1:1,000; these antibodies recognize HNP-1 to -3) or polyclonal rabbit anti-human antibodies against HBD-2 (Biologo, Kronshagen, Germany, no. DEF002; dilution, 1:100).

Techniques: Derivative Assay

Journal: Arthritis Research & Therapy

Article Title: Tumor necrosis factor and norepinephrine lower the levels of human neutrophil peptides 1-3 secretion by mixed synovial tissue cultures in osteoarthritis and rheumatoid arthritis

doi: 10.1186/ar3044

Figure Lengend Snippet: Influence of norepinephrine (NE) and cortisol (Cort) on human neutrophil peptides 1-3 (HNP1-3) levels in patients with rheumatoid arthritis (RA) and osteoarthritis (OA) . (a, b) Influence of norepinephrine on HNP1-3 levels in supernatants of mixed synoviocytes. The data are derived from five OA and five RA patients (two replicates of every patient). Control HNP1-3 values were as described in the legend of Figure 4. (c, d) Influence of cortisol or norepinephrine plus cortisol on HNP1-3 levels in supernatants of mixed synoviocytes. The data are derived from nine OA and seven RA patients (three replicates of every patient). Control HNP1-3 levels were 160.2 ± 50.3 pg/ml in OA patients and 73.5 ± 19.9 pg/ml in RA patients.

Article Snippet: Then, sections were incubated with either monoclonal mouse anti-human antibodies against HNP1-3 (BMA Biomedicals, Augst, Switzerland, no. T 1034; dilution, 1:1,000; these antibodies recognize HNP-1 to -3) or polyclonal rabbit anti-human antibodies against HBD-2 (Biologo, Kronshagen, Germany, no. DEF002; dilution, 1:100).

Techniques: Derivative Assay

Journal: Cureus

Article Title: Expression of Antimicrobial Peptides and Cytokines in Human Omentum Following Abdominal Surgery

doi: 10.7759/cureus.17477

Figure Lengend Snippet: (a) Immunoblot detection of anti-microbial proteins (LL-37, HNP1-3, HBD-1, HBD-2) and β-actin in clean-contaminated and contaminated omentum tissue as compared to control / naïve omentum tissue. (b) Densitometric analysis of Immunoblots for anti-microbial proteins (LL-37, HNP1-3, HBD-1, HBD-2) in clean-contaminated and contaminated omentum tissue as compared to control / naïve omentum tissue. β-Actin was used as an internal control to normalize the data. All the values represent mean± S.D (*p<0.05-significant, ***p<0.0001-highly significant). MOD-Mean Optical Density

Article Snippet: Immunoblotting was performed with primary antibodies (1:2000 rabbit anti-human LL37( Cat.no .A1640) polyclonal antibody, 1:2000 rabbit anti-human HNP1-3 ( Cat.no .AB37410) polyclonal antibody, and 1:2000 rabbit anti-human HBD2 ( Cat.no .A1643) polyclonal antibody from Immunotag, Geno Technology, St. Louis, US and 1:2000 rabbit anti-human HBD1 ( Cat.no .14738-1-AP) polyclonal antibody from Proteintech US, 1:2000 rabbit anti-human β-actin (Santa Cruz Biotechnology, USA)) at 40°C for overnight with gentle rocking.

Techniques: Western Blot

Journal: Cureus

Article Title: Expression of Antimicrobial Peptides and Cytokines in Human Omentum Following Abdominal Surgery

doi: 10.7759/cureus.17477

Figure Lengend Snippet: Specific primer sequences

Article Snippet: Immunoblotting was performed with primary antibodies (1:2000 rabbit anti-human LL37( Cat.no .A1640) polyclonal antibody, 1:2000 rabbit anti-human HNP1-3 ( Cat.no .AB37410) polyclonal antibody, and 1:2000 rabbit anti-human HBD2 ( Cat.no .A1643) polyclonal antibody from Immunotag, Geno Technology, St. Louis, US and 1:2000 rabbit anti-human HBD1 ( Cat.no .14738-1-AP) polyclonal antibody from Proteintech US, 1:2000 rabbit anti-human β-actin (Santa Cruz Biotechnology, USA)) at 40°C for overnight with gentle rocking.

Techniques: Sequencing