Evaluation of CD86 expression and MHC class II molecule internalization in THP-1 human monocyte cells as predictive endpoints for contact sensitizers.
The aim of this study was to explore the usefulness of a human monocyte cell line in the development of in vitro models for predictive testing of contact sensitizers. Several studies have shown that contact sensitizers induce CD86 expression and enhanced internalization of MHC class II molecules in dendritic cells (DCs). We used THP-1, a human monocyte cell line, as a replacement for DCs for evaluation of these phenotypical alterations as predictive endpoints for contact sensitizers. Known sensitizers and irritants were evaluated. After 24-h exposure to samples, the expression of CD86 on THP-1 cells was measured by flow cytometry. Sensitizers such as dinitrochlorobenzene (DNCB), 2-mercaptobenzothiazole (MBT), eugenol, p-phenylenediamine (PPDA) and ammonium tetrachloroplatinate (Pt) enhanced CD86 expression on THP-1 cells, while nickel sulfate, cobalt sulfate and irritants such as methylsalicylate (MS), sodium dodecyl sulfate (SDS) and dimethyl sulfoxide (DMSO) did not augment CD86 expression. A synergistic effect was observed when DNCB and IFN-alpha were added simultaneously to a culture of THP-1 cells. Furthermore, internalization of MHC class II molecules was observed when the cells were treated with some of sensitizers for 2 h. The inducing effects of chemicals on the two phenotypical alterations were the same. These results suggest that these test systems can be used to predict contact-sensitizing ability of chemicals as an in vitro sensitization assay.
Ashikaga
T
T
Shiseido Safety and Analytical Center, Safety Research Labs, 2-12-1 Fukuura, Kanazawa-ku, Yokohama-shi, 236-8643, Japan. takao.ashikaga@to.shiseido.co.jp
Hoya
M
M
Itagaki
H
H
Katsumura
Y
Y
Aiba
S
S
D000826:Animal Testing Alternatives; D015703:Antigens, CD / Q000096:biosynthesis*; D051940:B7-2 Antigen; D018929:Cell Culture Techniques; D003877:Dermatitis, Contact / Q000276:immunology* / Q000503:physiopathology; D004353:Drug Evaluation, Preclinical; D023381:Endpoint Determination; D005786:Gene Expression Regulation; D005802:Genes, MHC Class II; D006801:Humans; D007509:Irritants / Q000009:adverse effects; D008562:Membrane Glycoproteins / Q000096:biosynthesis*; D009000:Monocytes; D010641:Phenotype; D011237:Predictive Value of Tests
D015703:Antigens, CD; D051940:B7-2 Antigen; C497964:CD86 protein, human; D007509:Irritants; D008562:Membrane Glycoproteins