goat antihuman cxcl12  (R&D Systems)

Journal: Contact Dermatitis

Article Title: Titanium salts tested in reconstructed human skin with integrated MUTZ ‐3‐derived Langerhans cells show an irritant rather than a sensitizing potential

doi: 10.1111/cod.13666

Figure Lengend Snippet: MUTZ‐LC migration out of the epidermis after exposure to titanium(IV) bis(ammonium lactato)dihydroxide is CCL5 dependent. RHS‐LCs were unexposed (U), exposed to H 2 O vehicle (V), or 170 mM TiALH (+) for 24 hours. Chemical exposure was performed in the presence of neutralizing antibodies to CXCL12 (+) or CCL5 (+) or IgG1 isotype control (−). ( A ) Epidermal sheets isolated from RHS‐LCs stained with anti‐CD1a‐PE are shown. Fluorescence intensity (light) shows the presence of MUTZ‐LCs in the epidermal sheets. ( B ) CFSE/CD1a‐PE MUTZ‐LCs in the epidermal sheets were quantified using NIS‐Elements software. Data represent the average of four individual experiments performed in duplicate ± SEM. * P < .05 calculated using the Mann‐Whitney U test. Ig, immunoglobulin; LC, Langerhans cell; PE, phycoerythrin; RHS, reconstructed human skin; SEM, standard error of the mean; TiALH, titanium(IV) bis(ammonium lactato)dihydroxide

Article Snippet: For the blocking experiments, previously established optimal blocking concentrations of 7 μg/mL goat antihuman CCL5 (AF‐278‐NA; R&D systems, Minneapolis, Minnesota), 7 μg/mL goat antihuman CXCL12 (AF‐310‐NA, R&D systems), or 7 μg/mL polyclonal goat immunoglobulin G (IgG)G isotype antibody (6‐001‐F, R&D systems) were added to the culture medium 2 hours prior to chemical exposure.,

Techniques: Migration, Control, Isolation, Staining, Fluorescence, Software, MANN-WHITNEY

Journal: Contact Dermatitis

Article Title: Titanium salts tested in reconstructed human skin with integrated MUTZ ‐3‐derived Langerhans cells show an irritant rather than a sensitizing potential

doi: 10.1111/cod.13666

Figure Lengend Snippet: RHS dermis is CD68 + /IL‐10 high /CCR7 low /IL‐1β low after CCL5‐dependent MUTZ‐LC migration. RHS‐LCs were unexposed (U), exposed to H 2 O vehicle (V), 170 mM TiALH (+), or NiSO 4 (10 mM) for 24 hours. ( A ) Chemical exposure was performed in the presence of neutralizing antibodies to CXCL12 (+) or CCL5 (+) or IgG1 isotype control (−). CFSE/Langerin‐APC fluorescence intensity of MUTZ‐LCs in the dermis was quantified using the CellQuest Pro FACS analysis software. Real time‐polymerase chain reaction shows increased ( B ) IL‐1β and ( C ) CCR7 mRNA after NiSO 4 exposure, but not after titanium(IV) bis(ammonium lactato)dihydroxide exposure and ( D ) increased IL‐10 mRNA after exposure to titanium(IV) bis(ammonium lactato)dihydroxide but not after exposure to NiSO 4 . ( E ) Increased numbers of viable CD68 + cells (flow cytometry) in RHS‐LC dermis after exposure to titanium(IV) bis(ammonium lactato)dihydroxide but not after exposure to NiSO 4 . Data represent the average of four individual experiments performed in duplicate ± SEM. * P < .05 calculated using the Mann‐Whitney U test. CFSE, carboxyfluorescein succinimidyl ester; FACS, fluorescence‐activated cell sorting; Ig, immunoglobulin; IL, interleukin; LC, Langerhans cell; mRNA, messenger RNA; NiSO 4 , nickel sulfate; RHS, reconstructed human skin; SEM, standard error of the mean

Article Snippet: For the blocking experiments, previously established optimal blocking concentrations of 7 μg/mL goat antihuman CCL5 (AF‐278‐NA; R&D systems, Minneapolis, Minnesota), 7 μg/mL goat antihuman CXCL12 (AF‐310‐NA, R&D systems), or 7 μg/mL polyclonal goat immunoglobulin G (IgG)G isotype antibody (6‐001‐F, R&D systems) were added to the culture medium 2 hours prior to chemical exposure.,

Techniques: Migration, Control, Fluorescence, Software, Real-time Polymerase Chain Reaction, Flow Cytometry, MANN-WHITNEY, FACS