Babiuk C, Hastings K L, Dean J H
Chemical Industry Institute of Toxicology, Department of Cell Biology, Research Triangle Park, North Carolina 27709.
Fundam Appl Toxicol. 1987 Nov;9(4):623-34. doi: 10.1016/0272-0590(87)90169-2.
Ethylenediamine (EDA) is reported to be a poorly characterized iatrogenic and occupational contact sensitizer. To better characterize EDA hypersensitivity, a guinea pig model was employed in which the animals were exposed epicutaneously to simulate conditions of human exposure, and selected immune parameters were measured. Induction of hypersensitivity was by the Buehler occluded patch method (6 hr application/day, once a week for 3 consecutive weeks) to 10, 20, 30, or 40% EDA, using either an ethanol or acetone/corn oil vehicle. Fourteen days after the last induction, guinea pigs were challenged by patch application of 2% EDA (nonirritating). The incidence of responders for erythema in the 10% EDA (ethanol) treatment group was 83 and 50% at 24 and 48 hr, respectively. In the 10% EDA (acetone/corn oil) group the corresponding values were 50 and 17%. For 20, 30, and 40% EDA, in either vehicle, the incidence of erythema was 83 to 100%. Severity grades (scale = 0-3) for cutaneous reactions to increasing concentrations of EDA in ethanol ranged from 0.8 to 2.5; those for EDA in acetone/corn oil ranged from 0.6 to 2.8. Using an enzyme-linked immunosorbent assay developed to detect the predominant serum antibodies to EDA, it was shown that guinea pigs treated by patch application did not produce the main allergic antibody IgG specific for EDA. However, intradermal administration of an EDA-guinea pig serum albumin conjugate (EDA-GSA) to guinea pigs presensitized by patch application resulted in antibody production by 39 and 86% of the animals, at the initial and second dosing, respectively. An in vitro blastogenesis assay, using peripheral blood lymphocytes from EDA-sensitized guinea pigs, was developed to identify specific chemical allergens implicated in vivo sensitization. Maximum tritiated thymidine ([3H]TdR) incorporation by lymphocytes stimulated in vitro with EDA-GSA was observed on Day 7. Optimal antigen concentration for maximum lymphocyte proliferation ranged from 5 to 50 micrograms/ml, the major variation being attributable to interanimal differences. These results indicate that epicutaneous application of EDA in the guinea pig induces a Type IV delayed hypersensitivity; immunological memory to the hapten is maintained in cultured lymphocytes, suggesting the potential usefulness of the lymphocyte transformation test for in vitro diagnosis of chemically induced hypersensitivity in humans.
据报道，乙二胺（EDA）是一种特征不明的医源性和职业性接触致敏剂。为了更好地描述EDA超敏反应，采用了豚鼠模型，将动物进行皮肤暴露以模拟人类暴露情况，并测量选定的免疫参数。通过Buehler封闭贴片法（每天应用6小时，连续3周每周一次），使用乙醇或丙酮/玉米油载体，对10%、20%、30%或40%的EDA进行致敏。在最后一次致敏后14天，通过贴片应用2% EDA（无刺激性）对豚鼠进行激发。在10% EDA（乙醇）治疗组中，24小时和48小时时红斑反应者的发生率分别为83%和50%。在10% EDA（丙酮/玉米油）组中，相应的值分别为50%和17%。对于20%、30%和40%的EDA，无论使用何种载体，红斑发生率均为83%至100%。乙醇中EDA浓度增加时皮肤反应的严重程度分级（范围 = 0 - 3）为0.8至2.5；丙酮/玉米油中EDA的分级为0.6至2.8。使用开发的酶联免疫吸附测定法检测针对EDA的主要血清抗体，结果表明通过贴片应用处理的豚鼠不会产生针对EDA的主要过敏抗体IgG。然而，对通过贴片应用预先致敏的豚鼠皮内注射EDA - 豚鼠血清白蛋白偶联物（EDA - GSA），在初次给药和第二次给药时，分别有39%和86%的动物产生抗体。开发了一种体外增殖试验，使用来自EDA致敏豚鼠的外周血淋巴细胞，以鉴定体内致敏中涉及的特定化学过敏原。在第7天观察到用EDA - GSA体外刺激的淋巴细胞掺入氚化胸腺嘧啶核苷（[3H]TdR）的最大值。最大淋巴细胞增殖的最佳抗原浓度范围为5至50微克/毫升，主要差异归因于动物个体差异。这些结果表明，在豚鼠中经皮应用EDA可诱导IV型迟发型超敏反应；对半抗原的免疫记忆在培养的淋巴细胞中得以维持，这表明淋巴细胞转化试验在体外诊断人类化学诱导的超敏反应中可能具有实用性。
