KMID : 0379520030190020133
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Çѱ¹µ¶¼ºÇÐȸÁö 2003 Volume.19 No. 2 p.133 ~ p.139
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A Non-radioisotopic Endpoint Using Bromodeoxyuridine ELISA Method for Murine Local Lymph Node Assay
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Lee Jong-Kwon
Park Jae-Hyun Park Seung-Hee Kim Hyung-Soo Chung Seung-Tae Eom Joon-Ho Yun So-Mi Jang Eun-Jung Choi Kwang-Sik
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Abstract
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Allergic contact dermatitis may be caused by a wide variety of chemicals. A murine local lymph node assay (LLNA) has been developed as an alternative to guinea pig models for assessing the contact sensitization potential of chemical. However, there is a need to develop a nonradioisotopic endpoint for the LLNA, because of the radioisotopic method¡¯¡¯¡¯¡¯s requiring the use of special facilities. In this study, we investigated the development of a nonradioisotopic endpoint for LLNA using ELISA (enzyme-linked immunosorbent assay). Female Balb/c mice were treated by the topical application on the dorsum of both ears with four different strong sensitizers, 2,4-dinitrochlorobenzene (DNCB), oxazolone (OXZ), toluene diisocyanate (TDI), and trimellitic anhydride (TMA), and a strong irritant, sodium lauryl sulfate (SLS), once daily for three consecutive days. The proliferation of cells in the auricular Iymph node was analyzed by means of the labelling index (Ll) of bromodeoxyuridine (BrdU) incorporation into cells. The weights of the Iymph nodes in the mice treated with allergens, DNCB, OXZ, TDl and TMA were increased compared to the vehicle control. The stimulation index (Sl) of mice treated with DNCB, OXZ, TDl, and TMA was over three-fold increase compared to the vehicle control. However, the S1 of mice exposed to SLS was not significantly increased compared to the vehicle control, while the lymph node weight of SLS was significantly increased. These results suggest that the LLNA modified endpoint using ELISA based on BrdU incorporation could provide a useful method of screening for irritants and allergens.
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KEYWORD
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Local lymph node assay, BrdU, ELISA, 2, 4-Dinitrochlorobenzene, Toluene diisocyanate, Sodium lauryl sulfate
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