Preliminary study for integrating DPRA with h-CLAT to predict skin sensitizers / 中国实验动物学报

ABSTRACT

Objective To establish a detection method integrating DPRA ( direct peptide reactivity assay) with h?CLAT ( human cell line activation test) to screen the skin sensitization potency of chemicals and plant extracts. Methods 12 chemicals and 7 plant extracts were chosen as the test substances. Firstly, the test substances were incubated together with two different peptides ( cysteine and lysine) respectively for reaction for 24 h. The peptide consumptions were analyzed by HPLC. Simultaneously, THP?1 cells were cultured in vitro and then exposed to different concentrations of test sub?stances for 24 h to examine the cell viability, cell surface markers CD54 and CD86 were assessed by flow cytometry. The predicting results were compared further between DPRA and h?CLAT. Results 12 chemicals were distinguished correctly by DPRA classified as 2 non?sensitizers and 10 sensitizers. The results of DPRA were in accordance with h?CLAT. Predic?ting the sensitization potency of plant extracts by DPRA showed that 6 plant extracts were determined as suspected sensiti?zers except for green tea extract. But using the method of h?CLAT, 4 plant extracts were examined as suspected sensitizers except for green tea extract, herba portulacae extract and ginseng fruit extract. The coherence of DPRA and h?CLAT was 0?57. Conclusion This detection method integrating DPRA with h?CLAT can predict single compound accurately. As for complex compound, it can achieve preliminary prediction and need other integrating methods to make a further identifica?tion.