title: Comparison of a novel CXCL12/CCL5 dependent migration assay with CXCL8 secretion and CD86 expression for distinguishing sensitizers from non-sensitizers using MUTZ-3 Langerhans cells.
abstract: As the induction of contact hypersensitivity is the result of a series of cellular processes, including maturation and migration of epidermal dendritic cells (Langerhans cells (LC)), a battery of assays based on these in vivo events might provide a robust in vitro predictability model for distinguishing sensitizers from non-sensitizers. Therefore, assays with read-out for changes in CD86 expression and CXCL8 secretion were compared with a novel functional assay based on the in vitro migratory behaviour of LC. In all three assays LC derived from the human myeloid-leukaemia-cell-line MUTZ-3 (MUTZ-LC) were used. Exposure of MUTZ-LC to a panel of five sensitizers and three non-sensitizers resulted in increased CD86 expression in only 3/5 sensitizers, but also in 1/3 non-sensitizers. In contrast, CXCL8 secretion was uniformly increased after exposure to all sensitizers, but not after exposure to non-sensitizers. In a transwell migration assay, preferential migration of sensitizer-exposed MUTZ-LC towards CXCL12 was observed (5/5 sensitizers), whereas non-sensitizer-exposed MUTZ-LC only migrated towards CCL5 (3/3 non-sensitizers). In conclusion, the novel MUTZ-LC migration assay and analysis of CXCL8 secretion proved to be more successful than analysis of CD86 in predicting sensitizers from non-sensitizers and therefore warrant further investigation in the field of in vitro assay development.
authors.affiliation: Department of Dermatology, VU University Medical Centre, Amsterdam, The Netherlands.
authors.forename: Krista
authors.initials: K
authors.lastname: Ouwehand
authors.forename: Sander W
authors.initials: SW
authors.lastname: Spiekstra
authors.forename: Judith
authors.initials: J
authors.lastname: Reinders
authors.forename: Rik J
authors.initials: RJ
authors.lastname: Scheper
authors.forename: Tanja D
authors.initials: TD
authors.lastname: de Gruijl
authors.forename: Susan
authors.initials: S
authors.lastname: Gibbs
mesh_terms: D051940:B7-2 Antigen / Q000235:genetics / Q000378:metabolism*; D045744:Cell Line, Tumor; D002465:Cell Movement / Q000502:physiology*; D018946:Chemokine CCL5 / Q000378:metabolism*; D054377:Chemokine CXCL12 / Q000378:metabolism*; D003877:Dermatitis, Contact / Q000503:physiopathology*; D004353:Drug Evaluation, Preclinical; D005786:Gene Expression Regulation; D006801:Humans; D016209:Interleukin-8 / Q000235:genetics / Q000378:metabolism*; D007509:Irritants / Q000633:toxicity*
chemical_list: D051940:B7-2 Antigen; C483024:CCL5 protein, human; C497964:CD86 protein, human; C515255:CXCL12 protein, human; C508606:CXCL8 protein, human; D018946:Chemokine CCL5; D054377:Chemokine CXCL12; D016209:Interleukin-8; D007509:Irritants