title: Inter-laboratory study of the in vitro dendritic cell migration assay for identification of contact allergens.
abstract: The aim of this study was to investigate the transferability of technology and reproducibility of MUTZ-3 derived Langerhans Cell (MUTZ-LC) migration assay. The protocol was transferred from the NL-lab to two Sens-it-iv project partners (UK-lab, Italy-lab). Intra- and inter-laboratory variation with regards to MUTZ-3 progenitor culture, differentiation to MUTZ-LC, maturation and migration assay were investigated. In the transwell-migration-assay, preferential migration of sensitizer-exposed MUTZ-LC towards CXCL12 was observed (three sensitizers), whereas non-sensitizer-exposed MUTZ-LC only migrated towards CCL5 (two non-sensitizers). Four pre-pro-haptens were also identified by UK-lab. When taking the arbitrary criteria of at least two of three independent repetitions per laboratory having to have a CXCL12/CCL5 ratio>1.1 for classification as a sensitizer, all sensitizers tested in all labs were easily distinguished from all non-sensitizers. The number of repetitions giving false negative or false positive was very low (only 7 out of a total of 54 repetitions), indicating that both intra- and inter-laboratory variation was extremely low. Even though only a few chemicals were tested in this study, we show clearly that the in vitro DC migration assay is transferable between laboratories. The results were consistent between the laboratories, and the dose response data were reproduced in the three laboratories.
authors: Centre for Research in Biomedicine, Faulty of Health and Life Sciences, University of the West of England, Bristol, UK.
authors: B
authors: B
authors: Rees
authors: S W
authors: SW
authors: Spiekstra
authors: M
authors: M
authors: Carfi
authors: K
authors: K
authors: Ouwehand
authors: C A
authors: CA
authors: Williams
authors: E
authors: E
authors: Corsini
authors: J D
authors: JD
authors: McLeod
authors: S
authors: S
authors: Gibbs
mesh_terms: D000485:Allergens / Q000276:immunology*; D045744:Cell Line, Tumor; D054443:Cell Migration Assays; D018946:Chemokine CCL5 / Q000276:immunology; D054377:Chemokine CXCL12 / Q000276:immunology; D006241:Haptens / Q000276:immunology*; D006801:Humans; D007753:Laboratories; D007801:Langerhans Cells / Q000166:cytology / Q000276:immunology*; D015203:Reproducibility of Results; D018581:Technology Transfer
chemical_list: D000485:Allergens; C483024:CCL5 protein, human; C515255:CXCL12 protein, human; D018946:Chemokine CCL5; D054377:Chemokine CXCL12; D006241:Haptens