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Reconstitution of in vitro testing for metal allergy by means of LPT

利用 LPT 重建金属过敏体外测试

基本信息

批准号：
03557085
负责人：
FUJII Hiroyuki
金额：
$ 4.93万
依托单位：
NAGASAKI UNIVERSITY
依托单位国家：
日本
项目类别：
Grant-in-Aid for Developmental Scientific Research (B)
财政年份：
1991
资助国家：
日本
起止时间：
1991 至 1992
项目状态：
已结题

来源：
https://kaken.nii.ac.jp/grant/KAKENHI-PROJECT-03557085/
关键词：
metal allergy in vitro test lymphocyte prolifiration test 2-mercaptoethanol metal sensitized mice allergen bind fraction 金属アレルギ-

项目摘要

In order to reconstitution of in vitro testing (LPT:Lymphocyte Prolifiration Test) for metal allergy, following three series of investigation were made.1. Lymphocyte Prolifiration test: 1) Twelve highly positive reactors to patch test with Hg^<2+>, 5 highly positive reactors to patch test with Co^<2+> and 17 negative reactors to those, were participated in Lymphocyte Proliferation test(LPT). 2) A lymphocyte proliferation by Hg^<2+> was markedly depressed with 2-mercaptoethanol (2-ME) in the culture medium. 3) It was suggested that optimal incubation of metal sensitized lymphocyte was 144hs with Omol for Hg^<2+> and 120hs with 1.3mu-1.3muM for Co^<2+>.4) It was concluded that LPT might be applicable for sensitivity test to both Hg^<2+> and Co^<2+>, although further efforts should certainly be need to get results with gratified reliability in clinical purpose, especially in the former.2. Inducement of metal sensitized mice: Hg^<2+> sensitized mice(Balb/c) were successfully induced by subcutaneous or intraperitoneal injection with allergen-conbinant-epidermal cells.3. allergen-bind-fractions of the epidermal cell: After 24hs of application of 5% Hgcl^2 on skin surface of the mouse tail, amounts of Hg^<2+> bound with the microsome, with the mitochondria and with the lysosome of the epidermal cells were regarded as 40%, 33% and 23%, respectively.

为了重建金属过敏的体外检测方法(LPT：淋巴细胞增殖试验)，进行了以下三个系列的研究。淋巴细胞增殖试验：1)用Hg2+斑贴试验12例高阳性反应，用钴斑贴试验5例高阳性反应，17例阴性反应参加淋巴细胞增殖试验。2)在培养液中加入2-巯基乙醇(2-ME)，可显著抑制Hg2+诱导的淋巴细胞增殖。3)金属致敏淋巴细胞的最佳孵育时间为144h，金属致敏淋巴细胞的最佳孵育时间为144h，金属致敏淋巴细胞的最佳孵育时间为144h，金属致敏淋巴细胞的最佳孵育时间为144h，金属致敏淋巴细胞的最佳孵育时间为144h，金属致敏淋巴细胞的最佳孵育时间为144h，金属致敏淋巴细胞的最佳孵育时间为144h，金属致敏淋巴细胞的最佳孵育时间为1.3MU-1.3MU。金属致敏小鼠的诱导：HG^&lt；2+&gt；致敏小鼠(Balb/c)皮下或腹腔内注射致敏变应原结合的表皮细胞，均可成功致敏。表皮细胞变应原结合组分：5%HgCl2涂抹于小鼠尾部皮肤表面24小时后，与表皮细胞微粒体、线粒体和溶酶体的结合量分别为40%、33%和23%。