The immunotoxicity of dibutyl phthalate on the macrophages in mice

Abstract

Objective: Dibutyl phthalate (DBP), a widely used phthalate chemical, is commonly used as plasticizer. It is well known that DBP causes reproductive and developmental diseases, but the effect of DBP on the immune system remains to be determined. We assessed the effect of DBP on immune functions of murine macrophages, which constitute a key component in the immune response.

Materials and methods: Murine peritoneal exudate macrophages (PEMs) were treated with 0, 1, 5, 10, 50 or 100 μM DBP in vitro for 24 h and then the viability of PEMs were measured by flow cytometry (FCM) and trypan blue count. To investigate the effect of DBP on the functions of PEMs, we treated the PEMs with moderate dose of DBP (0, 1, 5 or 10 μM) in vitro for 24 h. The phenotypes, phagocytosis and cytokine production of PEMs were measured by FCM or real-time PCR. The immunogenicity and antigen presenting capacity of PEMs treated with DBP in vitro were assessed both by the mixed lymphocytereaction (MLR) in vitro assay and through the injection of exposed cells in mice by the delayed-type hypersensitivity (DTH) assay.

Results: High dose of DBP (50–100 μM) showed cytotoxicity on PEMs, whereas after the treatment with moderate dose of DBP (1–10 μM) in vitro, PEMs expressed low level of CD36, CD80 and MHC-II molecules, and showed significantly decreased phagocytosis on apoptotic cells and Escherichia coli. In addition, DBP treatment exhibited a decrease in the cytokine production, immunogenicity and antigen-presenting capacity of PEMs.

Conclusions: The present study shows the effects of DBP on macrophages, demonstrating immunogenicity and decreased antigen presentation in vitro.

• Antigen-presenting capacity
• cytokine production
• dibutyl phthalate
• immunotoxicity
• macrophages
• phagocytosis

Acknowledgements

The authors wish to thank Dr Yan Cheng for her kind review of the manuscript, Mr Wenchao Ai and Mr Guilin Cheng for their expert technical assistance, Mr Wenping Xie for his excellent laboratory management and Ms Yuan Cui for her outstanding animal husbandry.