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J.Health Sci., 47(3), 258-271, 2001
Immunotoxicological Evaluation of Environmental Chemicals Utilizing Mouse Lymphocyte Mitogenesis Test
Humitoshi Sakazaki,a Hitoshi Ueno,a Kaori Umetani,a Hideo Utsumi,b and Katsuhiko Nakamuro*, a
aDivision of Environmental Health, Faculty of Pharmaceutical Sciences, Setsunan University, Hirakata, Osaka 573-0101, Japan and bDepartment of Biophysics, Graduate School of Pharmaceutical Sciences, Kyushu University, Higashi-ku, Fukuoka 812-8582, Japan
To establish a useful and convenient bioassay for evaluating immunotoxicological effects of environmental chemicals, lymphocyte mitogenesis tests of about 255 chemicals and environmental water were performed. We determined the growth-inhibitory effect of the environmental chemicals on the mouse splenic lymphocyte mitogenesis using lipopolysaccharide and concanavalin A as the specific mitogen for B cells and T cells, and evaluated toxicity on humoral immunity and cell-mediated immunity, respectively. The DNA content of grown cells was determined by the novel ethidium bromide-fluorophotometry method, with 96-well microplates. Of the 255 chemicals tested, 173 chemicals showed inhibitory effects on the mitogenesis. The data were classified into four typical inhibition patterns from the dose-response curves. The chemicals were categorized into six groups in the respect of inhibition selectivity between B cell mitogenesis and T cell mitogenesis. Some chemicals, such as metallic compounds, showed nonspecific effects for B cell and T cell-mitogenesis, which was attributed to the cytotoxicity. The previous reports relating to immunotoxicity found about 123 chemicals; 78 chemicals (63.4%) were positive and one chemical (0.8%) was negative for both experimental results and reported results. In total, the concordance between experimental results and reported results is 64.2%. In the application of river water concentrates for the lymphocyte mitogenesis test, some of the samples inhibited both B cell and T cell mitogenesis at the tested volume of 7.2 ml. These results indicated that the lymphocyte mitogenesis test is applicable in estimating immunotoxicity of pollutants in environmental water as well as authentic environmental chemicals.
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