Evidence for the induction of apoptosis in thymocytes by 2,3,7,8-tetrachlorodibenzo-p-dioxin in vivo.
2,3,7,8-Tetrachlorodibenzo-p-dioxin (TCDD) is well known for its immunotoxic effects particularly on the thymus. The exact mechanism by which TCDD induces thymic atrophy is not clear. In the current study, we investigated whether TCDD triggers apoptosis in thymocytes, when administered in vivo, by using the TdT-mediated FITC-dUTP nick end labeling method and analyzing the cell flow cytometrically. Significant apoptosis was detected at 8-12 hr after the TCDD injection but not at 24 hr or beyond, up to 120 hr of study. Furthermore, the induction of apoptosis was confirmed using the JAM test in which thymocytes from TCDD-treated mice, labeled with [3H]thymidine, exhibited increased DNA fragmentation when compared to the controls. Similar to TCDD treatment, administration of dexamethasone (5 or 100 mg/kg) into C57BL/6 mice triggered apoptosis that was only detected at 12 hr after administration of the drug and not thereafter. When thymocytes from TCDD- or dexamethasone-treated mice were cultured in vitro for 24 hr, they exhibited marked increase in apoptosis when compared to the vehicle-treated controls. However, TCDD, when added to in vitro cultures of thymocytes, failed to trigger apoptosis. Together, our studies demonstrate that TCDD can induce apoptosis in thymocytes in vivo. This can be detected only at an early stage following TCDD administration, possibly because of rapid clearance of apoptotic cells by the phagocytic cells in vivo.
Kamath
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Department of Biomedical Sciences and Pathobiology, Virginia-Maryland Regional College of Veterinary Medicine, Blacksburg 24061, USA.
Xu
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H
Nagarkatti
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PS
Nagarkatti
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M
D000818:Animals; D017209:Apoptosis / Q000187:drug effects* / Q000502:physiology; D002452:Cell Count / Q000187:drug effects; D002478:Cells, Cultured; D053938:DNA Fragmentation / Q000187:drug effects; D004254:DNA Nucleotidyltransferases / Q000378:metabolism; D003907:Dexamethasone / Q000633:toxicity; D004305:Dose-Response Relationship, Drug; D005260:Female; D005434:Flow Cytometry; D051379:Mice; D008810:Mice, Inbred C57BL; D000072317:Polychlorinated Dibenzodioxins / Q000633:toxicity*; D013950:Thymus Gland / Q000187:drug effects* / Q000473:pathology
D000072317:Polychlorinated Dibenzodioxins; D003907:Dexamethasone; D004254:DNA Nucleotidyltransferases