Journal: Arthritis Research & Therapy

Article Title: Anti-type II collagen immune complex-induced granulocyte reactivity is associated with joint erosions in RA patients with anti-collagen antibodies

doi: 10.1186/s13075-015-0523-7

Figure Lengend Snippet: Functional reactivity against IgG as part of surface-bound collagen type II autoantibody (anti-CII) immune complex and polyclonal IgG. Amounts and functional reactivity against IgG as part of surface-bound collagen type II autoantibody (anti-CII) immune complex and polyclonal IgG directly coated to identical enzyme-linked immunosorbent assay (ELISA) plates. (a) ELISA reactivity of IgG against type II collagen (CII) aligned to known concentrations of directly coated IgG; these anti-CII levels (μg/ml) were thereafter used in the comparisons below. (b) Peripheral blood mononuclear cell-derived tumor necrosis factor alpha (ΤΝFα). (c) , (d) Polymorphonuclear granulocyte expression of CD66b and of CD16 respectively were compared for different concentrations of anti-CII and polyclonal IgG. Panels depict one representative result out of two to four experiments performed for each comparison. MFI, mean fluorescence intensity; OD, optical density.

Article Snippet: ELISA plates were coated with native human CII (10 μg/ml) (ELISA grade; Chondrex Inc.) at 4°C overnight.

Techniques: Functional Assay, Enzyme-linked Immunosorbent Assay, Derivative Assay, Expressing, Fluorescence

Journal: BMC Microbiology

Article Title: Molecular and virulence characteristics of an outer membrane-associated RTX exoprotein in Pasteurella pneumotropica

doi: 10.1186/1471-2180-11-55

Figure Lengend Snippet: The binding ability and hemagglutination activity of the rPnxIIIA . The binding ability of rPnxIIIA to the ECMs as determined by ELISA (A to D) and hemagglutination activity of the rPnxIIIA with sheep erythrocytes (E). Changes in the ability of different concentrations of rPnxIIIA to bind to the rat collagen type I (A), rat collagen type II (B), mouse collagen type IV (C), and mouse laminin (D), as determined by measuring the A620. Changes in the hemagglutination activity of different concentration of rPnxIIIA with sheep erythrocytes (E).

Article Snippet: In brief, a 96-well microtiter plate coated with rat collagen type I (BD BioCoat, BD) was used for the binding assay of rat collagen type I, and rat collagen type II (Chondrex, Redmond, WA, USA), mouse collagen type IV (BD), and mouse laminin (BD) were differently coated on a 96-well microplate (As one, Osaka, Japan) according to the manufacturer's instructions.

Techniques: Binding Assay, Activity Assay, Enzyme-linked Immunosorbent Assay, Concentration Assay

Journal: BMC Microbiology

Article Title: Molecular and virulence characteristics of an outer membrane-associated RTX exoprotein in Pasteurella pneumotropica

doi: 10.1186/1471-2180-11-55

Figure Lengend Snippet: Presence of PnxIIIA, binding ability, hemagglutination activity, and cytotoxicity of reference strains of P. pneumotropica . (A) Western blotting analysis of cell lysates (5 μg of total protein) of the reference strains by using anti-rPnxIIIA IgG. (B) The binding ability of the reference strains against to the rat collagen type I. A 1-way ANOVA determined that there were significant differences between the strains ( P < 0.05). The mean value of A490 of strain ATCC 35149 (numbered as 1) or CCUG 26453 (5) is significantly different from that of the other strains by determination of Duncan's multiple-range test ( P < 0.05). (C) Changes in hemagglutination activity of the reference strains with sheep erythrocytes. (D) Percentage of cytotoxicity determined by LDH release from the supernatant of J774A.1 cells cultured with reference strains of P. pneumotropica . A 1-way ANOVA determined that there were significant differences between the strains ( P < 0.05). The mean values of cytotoxicity (%) of strain ATCC 35149 (numbered as 1) or ATCC 12555 (2) and CCUG 36632 (6) are significantly different from that of the other strains by determination of Duncan's multiple-range test ( P < 0.05). All sections of numbers are represented as follows: 1, ATCC 35149; 2, ATCC 12555; 3, CCUG 26450; 4, CCUG 26451; 5, CCUG 26453; 6, CCUG 36632.

Article Snippet: In brief, a 96-well microtiter plate coated with rat collagen type I (BD BioCoat, BD) was used for the binding assay of rat collagen type I, and rat collagen type II (Chondrex, Redmond, WA, USA), mouse collagen type IV (BD), and mouse laminin (BD) were differently coated on a 96-well microplate (As one, Osaka, Japan) according to the manufacturer's instructions.

Techniques: Binding Assay, Activity Assay, Western Blot, Cell Culture