Smith Holly W, Winstead Colleen J, Stank Kristen K, Halstead Bartley W, Wierda Daniel
Eli Lilly and Company, 2001 W Main Street, Greenfield, IN 46140, USA.
Toxicology. 2003 Dec 15;194(1-2):129-45. doi: 10.1016/j.tox.2003.07.002.
The purpose of this study was to examine the predictive value of humoral and cellular immune parameters in determining the immunotoxic effects of the oral administration of azathioprine (AZA), cyclophosphamide (CY), or cyclosporin A (CsA) at doses of 25/17, 10, or 25 mg/kg per day, respectively, for 30 days in F344 female rats. The effect of these known immunosuppressive compounds on the immune response was assessed in a humoral model that consisted of the administration of nitrophenyl-chicken gamma globulin (NP-CgammaG) and keyhole limpet hemocyanin (KLH) antigens during immunosuppressive treatment and the measurement of resulting rat antigen-specific IgG and IgM, as well as total IgG, levels. Cellular assessment parameters were collected from the same groups of animals as the humoral parameters and included organ weights and cellularity, hematology, lymphocyte phenotype characteristics, spleen cell mitogen stimulation (T and B cell-dependent), splenic natural killer (NK) cell cytotoxicity, and bone marrow cellularity and lymphocyte phenotype differential. Although decreases in several of the cellular assay parameters were observed, the only functional assays to demonstrate a statistically significant immunosuppressive effect by all three immunosuppressive agents were the antigen-specific serum IgG levels. The primary (day 10; 15 days post-immunization) and secondary (day 25; 5 days post-rechallenge) nitrophenyl (NP) responses were significantly suppressed by > or =60%. The use of NP hapten provided consistent responses when analyzed with a sensitive, well developed, ELISA methodology. Absolute lymphocyte phenotyping and lymphocyte hematology were also predictive of T cell immunosuppression for all three compounds. The data presented herein suggests that these two parameters, NP-IgG humoral response and lymphocyte phenotyping, are sufficient for identifying immunosuppressive compounds.
本研究的目的是检测体液和细胞免疫参数在确定口服硫唑嘌呤（AZA）、环磷酰胺（CY）或环孢素A（CsA）对F344雌性大鼠免疫毒性作用方面的预测价值，给药剂量分别为每天25/17、10或25mg/kg，持续30天。在一个体液模型中评估了这些已知免疫抑制化合物对免疫反应的影响，该模型包括在免疫抑制治疗期间给予硝基苯基 - 鸡γ球蛋白（NP - CγG）和钥孔戚血蓝蛋白（KLH）抗原，并测量由此产生的大鼠抗原特异性IgG和IgM以及总IgG水平。细胞评估参数取自与体液参数相同的动物组，包括器官重量和细胞数量、血液学、淋巴细胞表型特征、脾细胞丝裂原刺激（T细胞和B细胞依赖性）、脾自然杀伤（NK）细胞细胞毒性以及骨髓细胞数量和淋巴细胞表型差异。虽然观察到几个细胞检测参数有所下降，但所有三种免疫抑制剂均显示出统计学显著免疫抑制作用的唯一功能检测是抗原特异性血清IgG水平。初次（第10天；免疫后15天）和二次（第25天；再次激发后5天）硝基苯基（NP）反应被显著抑制≥60%。当采用灵敏、成熟的ELISA方法分析时，NP半抗原的使用提供了一致的反应。绝对淋巴细胞表型分析和淋巴细胞血液学也可预测所有三种化合物对T细胞的免疫抑制作用。本文提供的数据表明，这两个参数，即NP - IgG体液反应和淋巴细胞表型分析，足以识别免疫抑制化合物。
