Department of Toxicogenomics, Maastricht University, Maastricht, The Netherlands.
Toxicology. 2013 Jul 5;309:91-9. doi: 10.1016/j.tox.2013.04.003. Epub 2013 Apr 25.
The aim of the present study was to determine the sensitivity of the developing immune system to ethanol (EtOH) after exposure from postnatal day (PND) 10 onward. Adult Wistar dams and litters were exposed to EtOH via drinking water (0, 0.25, 1.5, 2.75, 4, 5.25, or 6.5% (w/v) EtOH ad libitum) and drinking water exposure of the F1 was continued from weaning until sacrifice. Immune assessments were performed at postnatal days (PNDs) 21, 42, and 70. Furthermore, Keyhole Limpet Hemocyanin (KLH) specific immune responses were evaluated following subcutaneous immunizations on PNDs 21 and 35. EtOH exposure affected innate immune responses, such as LPS-induced NO-production by adherent splenocytes, as well as adaptive immune responses as represented by KLH-specific parameters. The most sensitive developmental parameters included effects on maternal and pup bodyweight with calculated BMDs of 4.0% and 4.3% EtOH, respectively. The most sensitive immune parameters were affected at dose levels lower than those affecting developmental parameters and included KLH-specific immune responses, LPS-induced NO production by adherent splenocytes, and IL-10 production by ConA stimulated splenocytes. Calculated BMDs for these parameters were between 0.01% and 0.1% EtOH. A comparison of the results of this juvenile study with an extended one-generation reproductive toxicity study revealed that the juvenile study design may result in a higher sensitivity related to differences in the exposure design. These findings demonstrate the relative sensitivity of the developing immune system for EtOH exposure, the additional value of assessing functional immune parameters, and the importance of the juvenile window in developmental immunotoxicity testing.
本研究旨在确定在产后第 10 天（PND）后暴露于乙醇（EtOH）后发育中免疫系统的敏感性。成年 Wistar 母鼠及其幼崽通过饮用水（0、0.25、1.5、2.75、4、5.25 或 6.5%（w/v）EtOH 自由摄取）暴露于 EtOH，并且 F1 的饮用水暴露从断奶持续到牺牲。在出生后第 21、42 和 70 天进行免疫评估。此外，在第 21 和 35 天进行皮下免疫接种后，评估了钥孔血蓝蛋白（KLH）特异性免疫反应。EtOH 暴露影响先天免疫反应，例如粘附性脾细胞中 LPS 诱导的 NO 产生，以及代表 KLH 特异性参数的适应性免疫反应。最敏感的发育参数包括对母体和幼崽体重的影响，分别计算出 4.0%和 4.3% EtOH 的 BMD。最敏感的免疫参数受影响的剂量水平低于影响发育参数的剂量水平，包括 KLH 特异性免疫反应、粘附性脾细胞中 LPS 诱导的 NO 产生以及 ConA 刺激的脾细胞中 IL-10 的产生。这些参数的计算 BMD 在 0.01%至 0.1% EtOH 之间。将这项青少年研究的结果与一项扩展的一代生殖毒性研究进行比较，结果表明青少年研究设计可能会由于暴露设计的差异而导致更高的敏感性。这些发现表明发育中免疫系统对 EtOH 暴露的相对敏感性，评估功能性免疫参数的额外价值，以及青少年窗口期在发育性免疫毒性测试中的重要性。
