DEVELOPMENT OF AN IN VITRO MODEL FOR PREDICTING THE CONTACT SENSITIVITY TO OCCUPATIONAL CHEMICALS

开发用于预测职业化学品接触敏感性的体外模型

• 批准号: 04670313
• 负责人: MATSUSHITA Toshio
• 金额: $ 1.41万
• 依托单位: KAGOSHIMA UNIVERSITY
• 依托单位国家: 日本
• 项目类别: Grant-in-Aid for General Scientific Research (C)
• 财政年份: 1992
• 资助国家: 日本
• 起止时间: 1992 至 1993
• 项目状态: 已结题
• 来源: https://kaken.nii.ac.jp/en/grant/KAKENHI-PROJECT-04670313/
• 关键词: Occupational chemicals; Contact sensitivity; Cross-reaction; Dose-response; Evaluation; In vitro model; DNCB; DNBS; 量一反応関係; 感作性; in vitro評価; リンパ球増殖反応; IL-2

Contact sensitizers are considered to lead to an immune response characteristic of Th1-type helper cells activation, and interleukin-2 (IL-2) is a representative cytokine secreted by Th1 cells. In this study, contact sensitivity to DNCB and the cross-sensitivity of DNBS with DNCB were evaluated using hapten-stimulated in vitro IL-2 production by the lymph node cells from mice contact sensitized with DNCB.For induction of in vitro IL-2 production, the lymph node cells from sensitized or control mice were cultured with the indicated concentrations of hapten. IL-2 activity in 48 hr culture supernatants was measured by its ability to stimulate IL-2-dependent CTLL-2 cell growths using MTT colorimetric assay and it was expressed as stimulation index. The following facts have emerged from our study.(1)DNCB in the form of hapten can induce hapten-specific in vitro IL-2 production by the lymph node cells from DNCB-sensitized mice.(2)DNCB-stimulated in vitro IL-2 production exhibited a maximal response on the 7th day after in vivo induction of contact sensitization.(3)There was a cross-reaction between DNCB and DNBS in in vitro IL-2 production.(4)DNCB-specific in vitro IL-2 production and the cross-reaction to DNBS showed the strong dose-response relationships.(5)There were goodcorrelations between DNCB-induced in vitro IL-2 production and DNCB-induced in vivo contact hypersensitivity as well as DNBS-induced in vitro IL-2 production and DNBS-elicited in vivo contact hypersensitivity. Thus, the present study suggests that hapten-induced in vitro IL-2 production by the murine lymph node cells may be a new in vitro model for predicting the contact sensitivity and their cross-sensitivity of chemicals.

接触致敏剂被认为导致Th 1型辅助细胞活化的免疫应答特征，白细胞介素-2（IL-2）是由Th 1细胞分泌的代表性细胞因子。本研究采用半抗原刺激DNCB接触致敏小鼠淋巴结细胞体外产生IL-2的方法，评价了DNCB对接触敏感性及DNBS与DNCB的交叉敏感性。采用MTT比色法测定48小时培养上清液中IL-2活性，以刺激IL-2依赖性CTLL-2细胞生长的能力为指标。从我们的研究中发现了以下事实。（1）半抗原形式的DNCB能诱导DNCB致敏小鼠淋巴结细胞产生半抗原特异性的IL-2。（2）DNCB刺激的体外IL-2产生在体内致敏后第7天达到最大。（3）DNCB和DNBS在体外IL-2产生中存在交叉反应。（4）DNCB特异性IL-2的体外产生及与DNBS的交叉反应均呈明显的剂量-反应关系。（5）DNCB诱导的体外IL-2产生与DNCB诱导的体内接触性变态反应以及DNBS诱导的体外IL-2产生与DNBS诱导的体内接触性变态反应之间存在良好的相关性。因此，本研究表明，在体外半抗原诱导的小鼠淋巴结细胞产生IL-2可能是一个新的体外模型，用于预测接触敏感性和化学品的交叉敏感性。

项目成果

Xu BH,Aoyama K,and Ueda A: "In vitro evaluation of contact sensitivity to DNCB using haptcn-stimulated in vitro interleukin-2 production by lymph node cells" Toxicology Methods. 3. 183-194 (1993)

Xu BH、Aoyama K 和 Ueda A：“使用 haptcn 刺激的淋巴结细胞体外产生白细胞介素 2 来体外评估对 DNCB 的接触敏感性”毒理学方法。

Xu Bll,Aoyama K,Matsushita T,and Ueda A: "An in vitro model for evaluating cross-sensitivily of DNBS with DNCB using hapten-stimulated in vitro 1L-2 production by murine lymph node cells" Life Science. (in preparation).

Xu Bll、Aoyama K、Matsushita T 和 Ueda A：“利用鼠淋巴结细胞在体外半抗原刺激产生 1L-2 来评估 DNBS 与 DNCB 交叉敏感性的体外模型”生命科学。

Xu BH,Aoyama K.,Matsushita T.,and Ueda A: "An in vitro model for evaluating cross-sensitivity of DNBS with DNCB using hapten-stimulated in vitro IL-2 production by murine lymph node cells" Life Science. (in preparation).

Xu BH、Aoyama K.、Matsushita T. 和 Ueda A：“使用半抗原刺激的小鼠淋巴结细胞体外产生 IL-2 来评估 DNBS 与 DNCB 交叉敏感性的体外模型”生命科学。

Matsushita T., Xu BH., and Aovama K: "Pathology and workplace's management of occupational allergic contact dermatitis" Occupational Health Journal. 17(5). in press (1994)

Matsushita T.、Xu BH. 和 Aovama K：“职业性过敏性接触性皮炎的病理学和工作场所管理”职业健康杂志。

Xu BH.,Aoyama K.,and Ueda A: "In vitro evaluation of contact sensitivity to DNCB using hapten-stimulated in vitro interleukin-2 production by lymph node cells" Toxicology Methods. 3. 183-194 (1993)

Xu BH.、Aoyama K. 和 Ueda A：“使用半抗原刺激的淋巴结细胞体外产生白介素 2 来体外评估对 DNCB 的接触敏感性”毒理学方法。