Characterization Of Thymic Atrophy and the Mechanism of Thymocyte Depletion after in Vivo Exposure to a Mixture Of Herbicides

Abstract

3,4-Dichloropropionanilide (propanil) and 2,4-dichlorophenoxyacetic acid (2,4-D) are two commonly used herbicides that are marketed as a chemical mixture. It was hypothesized that the interaction between these two herbicides, when administered as a mixture, would result in a greater effect on the immune system than the individual components of the mixture. The present study demonstrates in a murine model that a mixture of propanil and 2,4-D, when compared to single herbicide exposures, exacerbates decreases in thymocyte populations 2 d postexposure and inhibits the repopulation of T-cells in the thymus 7 d postexposure. Exposure to 150 mg herbicide/kg body weight of propanil or 2,4-D alone had no effect on thymus weight. In contrast, decreases in the ratio of thymus weight to body weight (TW:BW) occurred 2 d after treatment with the mixture of 150 mg propanil/kg body weight + 150 mg 2,4-D/kg body weight (150/150). Thymic atrophy was associated with a decrease in the double-positive thymocyte population (CD4+CD8+) and correlated with sera corticosterone levels from 600 to 1000 pg/ml. Therefore, the hypothesis was tested that glucocorticoids, induced after exposure to herbicides, were responsible for the thymic atrophy and depletion of thymocytes. However, similar levels of corticosterone were induced after exposure to 50, 100, or 150 mg propanil/kg body weight, and 50/50 or 100/100 mixture treatments, doses that did not produce thymic atrophy or cell loss. In addition, RU 486, a glucocorticoid receptor blocker, only partially abrogated the thymic atrophy in mice exposed to the 150/150 mixture of herbicides. These results suggest that glucocorticoids are only partially responsible for herbicide-induced thymic atrophy. This study demonstrates that the effects of exposure to a mixture of chemicals cannot always be predicted based on single exposure data and emphasizes the importance of mixture-based studies.

We thank Gerald R. Hobbs, PhD (Departments of Statistics and Community Medicine, West Virginia University), for assistance with statistical analysis, Cynthia Cunningham, PhD (Technical Director, Flow Cytometric Core Facility, West Virginia University), for assistance with flow cytometry, and Kathleen Brundage, PhD (Department of Microbiology, Immunology and Cell Biology, West Virginia University), for helpful discussion and reading of the article before publication.This work was supported by the National Institutes of Health, grants ES07460 and ES010953. The Flow Cytometric Core Facility is supported by the National Institutes of Health, grant RR16440.

Notes

We thank Gerald R. Hobbs, PhD (Departments of Statistics and Community Medicine, West Virginia University), for assistance with statistical analysis, Cynthia Cunningham, PhD (Technical Director, Flow Cytometric Core Facility, West Virginia University), for assistance with flow cytometry, and Kathleen Brundage, PhD (Department of Microbiology, Immunology and Cell Biology, West Virginia University), for helpful discussion and reading of the article before publication.This work was supported by the National Institutes of Health, grants ES07460 and ES010953. The Flow Cytometric Core Facility is supported by the National Institutes of Health, grant RR16440.