Kashima R, Okada J, Ikeda Y
Biological Science Laboratories, Kao Corporation, Tochigi, Japan.
Food Chem Toxicol. 1994 Sep;32(9):831-6. doi: 10.1016/0278-6915(94)90159-7.
The efficacy of a lymph node cell proliferation assay in the guinea pig as a first stage screening method of predicting sensitizing potentials of chemicals was studied by using several haptens. Animals were sensitized by a single 24-hr occlusive patch (24 cp), intradermal injection (id) and a combination of id and 24 cp, at a concentration used for guinea pig conventional contact hypersensitivity assay methods. Control animals were treated with vehicle(s) only. Suspensions of the lymph node cells (LNC) were individually prepared and cultured with [3H]methyl thymidine ([3H]TdR). [3H]TdR incorporation was measured and a stimulation index (SI) was calculated as a ratio of the mean [3H]TdR incorporation in sensitized animals and the mean [3H]TdR incorporation in control animals. LNC sensitized by 24 cp with 2,4-dinitrochlorobenzene proliferated maximally and significantly at day 5, whereas this occurred at day 7 after id sensitization. Significant LNC proliferation and high SI values were obtained successively by a combination of 24 cp and id. Moreover, strongly sensitizing chemicals increased significant LNC proliferation (SI > 2.0); weakly to moderately sensitizing chemicals also induced significant LNC proliferation (SI = 1.3-1.7). On the other hand, a primary irritant, sodium dodecyl sulfate, failed to encourage LNC proliferation (SI approximately 1.0).
通过使用几种半抗原，研究了豚鼠淋巴结细胞增殖试验作为预测化学物质致敏潜力的第一阶段筛选方法的有效性。动物通过单次24小时封闭贴片（24cp）、皮内注射（id）以及id和24cp联合的方式进行致敏，致敏浓度采用豚鼠常规接触性超敏反应试验方法所使用的浓度。对照动物仅用赋形剂处理。分别制备淋巴结细胞（LNC）悬液，并与[3H]甲基胸腺嘧啶核苷（[3H]TdR）一起培养。测量[3H]TdR掺入量，并计算刺激指数（SI），其计算方法为致敏动物中[3H]TdR平均掺入量与对照动物中[3H]TdR平均掺入量之比。用2,4 -二硝基氯苯进行24cp致敏的LNC在第5天增殖达到最大且显著，而经id致敏的LNC在第7天才出现这种情况。通过24cp和id联合的方式依次获得了显著的LNC增殖和高SI值。此外，强致敏性化学物质可使LNC显著增殖（SI>2.0）；弱至中度致敏性化学物质也可诱导LNC显著增殖（SI = 1.3 - 1.7）。另一方面，一种原发性刺激物十二烷基硫酸钠未能促进LNC增殖（SI约为1.0）。
