Chronic TCDD exposure results in the dysregulation of gene expression in splenic B-lymphocytes and in the impairments in T-cell and B-cell differentiation in mouse model

2,3,7,8-Tetrachlorodibenzo-p-dioxin（TCDD） exposure in humans is associated with marked immune suppressions and increased incidence of lymphoblastic diseases.To elucidate mechanisms of impairments in humoral immune responses,we used a murine model.Following a 20-week administration of low doses of TCDD,we observed severely reduced antibody titers,dramatically decreased number of splenic Th1 and Th2 cells and an increase in CD19^＋ B cells.Transcriptional profiling of CD19^＋ B cells showed that markers of pre-B cells were significantly elevated,indicating delayed B cell maturation.These changes in B cells were accompanied by decreases of T helper cell numbers and reduced IgM and IgG titers.A transcriptome analysis of splenic B cells followed by Ingenuity Pathway Analysis（IPA） revealed a set of differentially expressed genes known to play roles in tumorigenesis,cell-proliferation and cell-migration.The most up-regulated transcript gene was Eph receptor A2（EphA2）,a known oncogene,and the most down-regulated transcript was ZBTB16 that codes for a negative transcriptional regulator important in epigenetic chromatin remodeling.IPA identified cAMP-responsive element modulator（CREM） and cAMP-responsive element binding protein 1（CREBl） as top upstream regulators.Consistently,a MAPPER promoter database analysis showed that all top dysregulated genes had CREM and/or CREBl binding sites in their promoter regions.In summary,our data showed that chronic TCDD exposure in mice caused suppressed humoral immunity accompanied with profound dysregulation of gene expression in splenic B-lymphocytes,likely through cAMP-dependent pathways.This dysregulation resulted in impairments in T-cell and B-cell differentiation and activation of the tumorigenic transcription program.

Development and characterization of monoclonal antibodies against human aryl hydrocarbon receptor

Aryl hydrocarbon receptor（Ah R）, a ligand-dependent nuclear receptor, is involved in a diverse spectrum of biological and toxicological effects. Due to the lack of three dimensional（3D）crystal or nuclear magnetic resonance structure, the mechanisms of these complex effects of AhR remain to be unclear. Also, commercial monoclonal antibodies（mA bs） against human AhR protein（h Ah R）, as alternative immunological tools, are very limited. Thus, in order to provide more tools for further studies on h Ah R, we prepared two m Abs（1D6 and 4A6） against h Ah R. The two newly generated m Abs specifically bound to amino acids 484–508（located in transcription activation domain） and amino acids 201–215（located in Per-ARNT-Sim domain）of h Ah R, respectively. These epitopes were new as compared with those of commercial m Abs.The m Abs were also characterized by enzyme-linked immunosorbent assay, western blot,immunoprecipitation and indirect immunofluorescence assay in different cell lines. The results showed that the two m Abs could recognize the linearized AhR s in six different human cell lines and a rat hepatoma cell line, as well as the h Ah R with native conformations. We concluded that the newly generated m Abs could be employed in AhR-based bioassays for analysis of environmental contaminants, and held great potential for further revealing the spatial structure of AhR and its biological functions in future studies.