Elucidation of cellular targets responsible for tetrachlorodibenzo-p-dioxin (TCDD)-induced suppression of antibody responses: I. The role of the B lymphocyte

doi:10.1016/0162-3109(88)90005-7

Immunopharmacology

Volume 16, Issue 3, November–December 1988, Pages 167-180

Immunopharmacology

https://doi.org/10.1016/0162-3109(88)90005-7 Get rights and content

Abstract

The objective of these studies was to identify the primary cellular target(s) responsible for TCDD (tetrachlorodibenzo-p-dioxin)-induced suppression of antibody production. Responses to T-independent and T-dependent antigens (administered in vivo or directly to splenic culture) were suppressed in a dose-related fashion in female B6C3F1 mice dosed for 5 consecutive days with 0.1, 1.0 and 10.0 μg/kg of 2,3,7,8-TCDD. When nonadherent (B and T) and adherent (macrophage) cells from vehicle- and TCDD-treated mice were combined in various combinations and immunized with either DNP-Ficoll (T-independent) or sRBCs (T-dependent), it was demonstrated that nonadherent cells, but not adherent cells, were functionally affected by TCDD. Similarly, as various combinations of B + macrophage and T + macrophage populations were immunized with sRBCs, the B cell was shown to be the primary target. Using LPS as the stimulus, an inhibition of the antibody response with no effect on the mitogenic response further indicated that the primary target of the TCDD-induced suppression of IgM antibody production is the B lymphocyte at the level of cell differentiation.

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Cited by (84)

TCDD attenuates EAE through induction of FasL on B cells and inhibition of IgG production
2021, Toxicology
Previously we demonstrated that 2,3,7,8-tetrachlorodibenzo-p-dioxin (TCDD) suppressed experimental autoimmune encephalomyelitis (EAE), a model to study multiple sclerosis (MS), through induction of regulatory T cells (Tregs) and suppression of effector T cell function in the spleen. Since B cells and specifically regulatory B cells (Bregs) have been shown to be so critical in the pathology associated with EAE and MS, we wanted to determine whether TCDD could also induce Bregs. We specifically hypothesized that a Fas ligand (FasL)+ Breg population would be induced by TCDD in EAE thereby triggering apoptosis in Fas-expressing effector T cells as one mechanism to account for inhibition of T cell function by TCDD. TCDD (0.1–2.5 μg/kg/day administered orally for 12 days) modestly increased the percentage of FasL + B cells in the spleen and spinal cord in TCDD-treated EAE mice. However, we did not detect significant increases in percentages of FasL + B cells using TCDD in vitro in mouse splenocytes or human peripheral blood mononuclear cells (PBMCs). Part of the modest effect by TCDD was likely related to the localized expression of FasL; for instance, in the spleen, FasL was more highly expressed by IgM^hiIgD^lo marginal zone (MZ) B cells, but IgM^loIgD^hi follicular (FO) B cells were more responsive to TCDD. Consistent with our observation of modest upregulation of FasL, we also observed modest changes in mitochondrial membrane potential in T cells co-cultured with isolated total B cells or IgM-depleted (i.e., FO-enriched) B cells from TCDD-treated EAE mice. These data suggest that while small microenvironments of apoptosis might be occurring in T cells in response to TCDD-treated B cells, it is not a major mechanism by which T cell function is compromised by TCDD in EAE. TCDD did robustly suppress IgG production systemically and in spleen and spinal cord B cells at end stage disease. Thus, these studies show that TCDD’s primary effect on B cells in EAE is compromised IgG production but not FasL + Breg induction.
The Aryl Hydrocarbon Receptor and Immunity
2018, Comprehensive Toxicology: Third Edition
The aryl hydrocarbon receptor (AhR), originally discovered as a xenobiotic receptor involved in upregulating metabolic enzymes, has become increasingly linked to physiological processes such as cell cycle regulation and cellular differentiation as well as disease states associated with these processes. The immune system is composed of numerous effector cells and protein mediators with specialized functions, all of which are highly dependent on cell cycle regulation and cellular differentiation and therefore offer a plethora of potential targets for modulation by the AhR. Indeed, the vast majority of studies support a direct or indirect influence of the AhR in most aspects of immunity. The focus of this chapter is to discuss these studies. Although AhR activation by dioxin and nondioxin ligands is a primary method for evaluating AhR-mediated effects, there is the possibility for off-target effects by AhR ligands (i.e., not AhR mediated) or for effects that do not accurately reflect the physiological function of the AhR (i.e., exogenous vs. endogenous ligands). Therefore, more emphasis will be given to immune effects that have been demonstrated to be AhR dependent by either the use of a chemical AhR antagonist or the knockdown or knockout of the AhR.
Comparative analysis of TCDD-induced AhR-mediated gene expression in human, mouse and rat primary B cells
2017, Toxicology and Applied Pharmacology
Citation Excerpt :
Additionally, the window of sensitivity during which TCDD can suppress IgM secretion is similar between mouse, human and rat B cells, suggesting a common mechanism of action of IgM suppression. Specifically, TCDD must be added to the activated B cells within the initial 12 h post-stimulation to suppress IgM secretion (Dooley and Holsapple, 1988; Sulentic et al., 1998; Kovalova et al., 2016). The relatively narrow window of sensitivity suggests TCDD-mediated interference with a critical event shortly following B-cell activation.
2,3,7,8-Tetrachlorodibenzo-p-dioxin (TCDD) is a persistent environmental pollutant that activates the aryl hydrocarbon receptor (AhR) resulting in altered gene expression. In vivo, in vitro, and ex vivo studies have demonstrated that B cells are directly impaired by TCDD, and are a sensitive target as evidenced by suppression of antibody responses. The window of sensitivity to TCDD-induced suppression of IgM secretion among mouse, rat and human B cells is similar. Specifically, TCDD must be present within the initial 12 h post B cell stimulation, indicating that TCDD disrupts early signaling network(s) necessary for B lymphocyte activation and differentiation. Therefore, we hypothesized that TCDD treatment across three different species (mouse, rat and human) triggers a conserved, B cell-specific mechanism that is involved in TCDD-induced immunosuppression. RNA sequencing (RNA-Seq) was used to identify B cell-specific orthologous genes that are differentially expressed in response to TCDD in primary mouse, rat and human B cells. Time course studies identified TCDD-elicited differential expression of 515 human, 2371 mouse and 712 rat orthologous genes over the 24-h period. 28 orthologs were differentially expressed in response to TCDD in all three species. Overrepresented pathways enriched in all three species included cytokine-cytokine receptor interaction, ECM-receptor interaction, focal adhesion, regulation of actin cytoskeleton and pathways in cancer. Differentially expressed genes functionally associated with cell-cell signaling in humans, immune response in mice, and oxidation reduction in rats. Overall, these results suggest that despite the conservation of the AhR and its signaling mechanism, TCDD elicits species-specific gene expression changes.
SHP-1 is directly activated by the aryl hydrocarbon receptor and regulates BCL-6 in the presence of 2,3,7,8-tetrachlorodibenzo-p-dioxin (TCDD)
2016, Toxicology and Applied Pharmacology
Citation Excerpt :
Among the myriad toxicities elicited by TCDD, immune toxicity, specifically, suppression of B cell activation and the primary humoral immune response represent a highly sensitive endpoint of TCDD exposure (Holsapple et al., 1991). Several studies have contributed towards identification of the molecular targets underlying TCDD-mediated suppression of the primary humoral immune response in rodents with B cell as the direct target of TCDD (Dooley and Holsapple, 1988). The specific process that TCDD impairs is the ability of B cells to differentiate into IgM-secreting plasma cells (Morris et al., 1993; Sulentic et al., 1998).
The environmental contaminant 2,3,7,8-tetrachlorodibenzo-p-dioxin (TCDD), which is a strong AHR agonist, causes significant suppression of human B cell activation and differentiation. The current studies describe the identification of Src homology phosphatase 1 (SHP-1) encoded by the gene PTPN6 as a putative regulator of TCDD-mediated suppression of B cell activation. Shp-1 was initially identified through a genome-wide analysis of AHR binding in mouse B cells in the presence of TCDD. The binding of AHR to the PTPN6 promoter was further confirmed using electrophoretic mobility shift assays in which, specific binding of AHR was detected at four putative DRE sites within PTPN6 promoter. Time-course measurements performed in human B cells highlighted a significant increase in SHP-1 mRNA and protein levels in the presence of TCDD. The changes in the protein levels of SHP-1 were also observed in a TCDD concentration-dependent manner. The increase in SHP-1 levels was also seen to occur due to a change in early signaling events in the presence of TCDD. We have shown that BCL-6 regulates B cell activation by repressing activation marker CD80 in the presence of TCDD. TCDD-treatment led to a significant increase in the double positive (SHP-1^hi BCL-6^hi) population. Interestingly, treatment of naïve human B cells with SHP-1 inhibitor decreased BCL-6 protein levels suggesting possible regulation of BCL-6 by SHP-1 for the first time. Collectively, these results suggest that SHP-1 is regulated by AHR in the presence of TCDD and may, in part through BCL-6, regulate TCDD-mediated suppression of human B cell activation.
Role of aryl hydrocarbon receptor polymorphisms on TCDD-mediated CYP1B1 induction and IgM suppression by human B cells
2016, Toxicology and Applied Pharmacology
Citation Excerpt :
Suppression of the immune system had been described in many animal species and is one of the most sensitive consequences of TCDD exposure (Holsapple et al., 1991; Sulentic and Kaminski, 2011). Moreover, TCDD had been shown to directly and specifically target B cell function (Dooley and Holsapple, 1988). Studies with AhR-deficient experimental models, including mice, demonstrated that most, if not all, of the toxic effects of dioxins are mediated by the aryl hydrocarbon receptor (AhR) (Harrill et al., 2015; Mimura and Fujii-Kuriyama, 2003).
Previous studies have demonstrated that most of the intraspecies variation in sensitivity to the toxic effects of 2,3,7,8-tetrachlorodibenzo-p-dioxin (TCDD), including suppression of antibody responses, in murine models is due to single nucleotide polymorphisms (SNPs) within the aryl hydrocarbon receptor (AhR) gene. The underlying reason for variation in sensitivity to TCDD-induced suppression of IgM responses among humans is not well understood, but is thought, in part, to be a result of different polymorphic forms of the AhR expressed by different individuals. In this study, the functional properties of six (P517S, R554K, V570I, V570I + P517S, R554K + V570I and P517S + R554K + V570I) human AhR variants were examined in the human B cell line, SKW 6.4. TCDD-induced Cyp1B1 and Cyp1A2 mRNA expression levels and Cyp1B1-regulated reporter gene activity, used for comparative purposes, were markedly lower in SKW cells containing the R554K SNP than in SKW-AHR⁺ (control AhR) cells. Furthermore, all AhR variants were able to mediate TCDD-induced suppression of the IgM response; however, a combined P517S + R554K + V570I variant partially reduced sensitivity to TCDD-mediated suppression of IgM secretion. Collectively, our findings show that the R554K human AhR SNP alone altered sensitivity of human B cells to TCDD-mediated induction of Cyp1B1 and Cyp1A2. By contrast, attenuation of TCDD-induced IgM suppression required a combination of all three SNPs P517S, R554K, and V570I.
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
2016, Journal of Environmental Sciences (China)
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 (CREB1) as top upstream regulators. Consistently, a MAPPER promoter database analysis showed that all top dysregulated genes had CREM and/or CREB1 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.

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Elucidation of cellular targets responsible for tetrachlorodibenzo-p-dioxin (TCDD)-induced suppression of antibody responses: I. The role of the B lymphocyte

Abstract

Biochem Biophys Res Commun

Cell Immunol.

Cell

Cell Immunol.

Biochem Biophys Res Commun

J Biol Chem

Immunopharmacology

Toxicol Appl Pharmacol

Immunopharmacology

Biochem Biophys Res Commun

Biochem Biophys Res Commun

Cell Immunol

Immunol Today

Chem Biol Interact

Toxicol Appl Pharmacol

Toxicology

Inositol trisphosphate and diacylglycerol as second messengers

Biochem J

Molecular mechanisms of transmembrane signaling in B lymphocytes

Annu Rev Immunol

The effects of mercaptoethanol and of peritoneal macrophages on the antibody-forming capacity of non-adherent mouse spleen cells in vitro

J Exp Med

Enhanced suppressor cell activity as a mechanism of immunosuppression by 2,3,7,8-tetrachlordibenzo-p-dioxin

Dose response, time-course and mechanism for suppression of cytotoxic T cell generation by 2,3,7,8-tetrachlorodibenzo-p-dioxin

The role of protein phosphorylation in neural and hormonal control of cellular activity

Nature