Abstract
The susceptibility of developing immune system to chemical disruption warrants the assessment of immune parameters in reproductive and developmental testing protocols. In this study, a wide range of immune endpoints was included in an extended one-generation reproduction toxicity study (EOGRTS) design to determine the relative sensitivity of immune and developmental parameters to ethanol (EtOH), a well-known developmental toxicant with immunomodulatory properties. Adult Wistar rats were exposed to EtOH via drinking water (0, 1.5, 4, 6.5, 9, 11.5 and 14 % (w/v EtOH)) during premating, mating, gestation and lactation and continuation of exposure of the F1 from weaning until killed. Immune assessments were performed at postnatal days (PNDs) 21, 42 and 70. Keyhole limpet hemocyanin (KLH)-specific immune responses were evaluated following subcutaneous immunizations on PNDs 21 and 35. EtOH exposure affected innate as well as adaptive immune responses. The most sensitive immune parameters included white blood cell subpopulations, ConA-stimulated splenocyte proliferation, LPS-induced NO and TNF-α production by adherent splenocytes and KLH-specific immune responses. Most parameters showed recovery after cessation of EtOH exposure after weaning in the 14 % exposure group. However, effects on LPS-induced NO and TNF-α production by adherent splenocytes and KLH-specific parameters persisted until PND 70. The results demonstrate the relative sensitivity to EtOH of especially functional immune parameters and confirm the added value of immune parameters in the EOGRTS. Furthermore, this study identified an expanded KLH-specific parameter set and LPS-induced NO and TNF-α production by adherent splenocytes as valuable parameters that can provide additional information on functional immune effects.
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Acknowledgments
The authors gratefully acknowledge the excellent technical assistance of Gerard van Beek and Lidy van Oostrum (and colleagues) and Aart Verhoef for animal handling and necropsy skills, Anja Dijkstra for help with the planning and logistics involved in this study and Eric Gremmer, Liset de la Fonteyne, Jolanda Vermeulen, Arja de Klerk, Bert Verlaan and Henny Verharen for immunologic evaluations.
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The authors declare that they have no conflict of interest.
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Tonk, E.C.M., de Groot, D.M.G., Wolterbeek, A.P.M. et al. Developmental immunotoxicity of ethanol in an extended one-generation reproductive toxicity study. Arch Toxicol 87, 323–335 (2013). https://doi.org/10.1007/s00204-012-0940-1
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DOI: https://doi.org/10.1007/s00204-012-0940-1