Effects of trichothecene structure on cytokine secretion and gene expression in murine CD4+ T-cells.

The effects of trichothecene structure on cytokine secretion and gene expression were assessed in primary CD4+ T-cells from murine spleen. CD4+ T-cells were stimulated with concanavalin A (Con A) for 2 or 7 days in the presence of various concentrations of the trichothecenes, vomitoxin (VT or deoxynivalenol), nivalenol (NIV), 15-acetyl deoxynivalenol (15-ADON), 3-acetyl deoxynivalenol (3-ADON), T-2 toxin (T-2) and verrucarin A (Ver A). Culture supernatants were subsequently analyzed for interleukin (IL)-2, IL-4 and IL-5 by ELISA.

Induction of cytokine mRNAs in mice after oral exposure to the trichothecene vomitoxin (deoxynivalenol): relationship to toxin distribution and protein synthesis inhibition.

The effects of oral exposure to 0, 5, and 25 mg/kg body wt vomitoxin (VT) on cytokine mRNA levels in spleen, Peyer's patches (PP), liver, kidney, and small intestine were evaluated in B6C3F1 mice at 2 and 4 hr postexposure using RT-PCR in conjunction with Southern hybridization analysis. The abundance of mRNAs for several cytokines was increased in VT-exposed mice with maximal effects occurring in the 25 mg/kg group at 2 hr. Specifically, IL-1 beta and IL-6 mRNA levels increased in spleen and PP following exposure to VT.

Comparative Detection of Fumonisin by HPLC, ELISA, and Immunocytochemical Localization in Fusarium Cultures.

Fumonisins are a group of mycotoxins that are elaborated by Fusarium moniliforme and Fusarium proliferatum and that have recently been associated with animal and human disease. In this study, the time course of fumonisin B (FB) production in corn was monitored in five Fusarium cultures using high-performance liquid chromatography (HPLC), enzyme-linked immunosorbent assay (ELISA), and in situ localization by an enzyme-linked immunocytochemical technique (ELICT). Using HPLC on culture extracts prepared with 50% (vol/vol) acetonitrile in water, FB was detectable at 3 days with maximal FB (ranging from 230 to 3,000 ppm) occurring between 14 and 28 days.

Effects of vomitoxin (deoxynivalenol) and cycloheximide on IL-2, 4, 5 and 6 secretion and mRNA levels in murine CD4+ cells.

The effects of continuous in vitro exposure to the trichothecene, vomitoxin (VT) or another protein synthesis inhibitor, cycloheximide (CHX), on interleukin (IL) secretion and mRNA levels were evaluated in murine splenic CD4+ cells. Significant increases were seen in supernatant IL-2, IL-4 and IL-5 obtained from 7 day Concanavalin A (Con A)-stimulated CD4+ cultures containing VT concentrations of 250, 100 and 100 ng/ml, respectively, compared with controls run in the absence of VT. The effect of VT on CD4+ cell proliferation was also assessed after culturing for 3, 5 and 7 days with Con A.

Effects of dihydrotestosterone and estradiol on experimental IgA nephropathy induced by vomitoxin.

Ingestion of the trichothecene vomitoxin (VT) by mice induces effects that mimic the common human glomerulonephritis, IgA nephropathy (IgAN). These include elevation of serum IgA, IgA immune complexes, and mesangial IgA deposition. Based on previous observations that male mice are more prone to VT-induced IgAN, the effects of castration of male and female B6C3F1 mice and sex hormone supplementation on several immunopathologic indicators of the disease were compared.

Flow cytometric analysis of the effects of in vitro exposure to vomitoxin (deoxynivalenol) on apoptosis in murine T, B and IgA+ cells.

The immunotoxic effects of the trichothecene vomitoxin (VT or deoxynivalenol) and other trichothecenes may be mediated by direct interaction with lymphocytes. In this study, flow cytometric cell cycle analysis was used in conjunction with phenotypic staining by specific fluorescein isothiocyanate antibody conjugates to assess the in vitro effects of VT and another protein synthesis inhibitor, cycloheximide (CHX), on apoptosis in specific T- and B-cell subsets within thymus, spleen and Peyer's patch (PP) cultures. Both VT and CHX markedly inhibited T-cell apoptosis in dexamethasone (9 alpha-fluoro-16 alpha-methylprednisolone)-induced (DEX+) cells isolated from thymus, spleen and PP.

Vomitoxin (deoxynivalenol)-induced IgA nephropathy in the B6C3F1 mouse: dose response and male predilection.

Oral exposure to the trichothecene vomitoxin (VT or deoxynivalenol) in mice induces marked elevation of total and autoreactive IgA, IgA immune complexes, and mesangial IgA deposition in a manner that is highly analogous to human IgA nephropathy. In this study, immunopathologic markers indicative of IgA nephropathy were compared in male and female B6C3F1 mice fed semipurified AIN-76A diet containing 0, 2, 10 or 25 ppm VT for 12 weeks. Males fed 10 and 25 ppm VT and females fed 25 ppm VT had increased serum IgA at 4 weeks.

Role of gender and strain in vomitoxin-induced dysregulation of IgA production and IgA nephropathy in the mouse.

Prolonged dietary exposure of female B6C3F1 mice to the trichothecene vomitoxin results in hyperproduction of immunoglobulin A (IgA) with a concurrent immunopathology that mimics human IgA nephropathy. To assess the role of gender and strain in the mouse model, semipurified AIN-76A diet containing 25 ppm vomitoxin was fed to B6C3F1 male mice and to B6C3F1, BALB/c, C3H/HeN, C3H/HeJ, and C57BL/6 female mice for 8 wk, and immunopathologic indicators of IgA nephropathy were compared to mice fed clean diet. At the cessation of the experiment, all treatment groups weighed less than respective controls.

Elevated gene expression and production of interleukins 2, 4, 5, and 6 during exposure to vomitoxin (deoxynivalenol) and cycloheximide in the EL-4 thymoma.

Aberrant elevation of serum IgA and induction of murine IgA nephropathy following dietary exposure to the naturally occurring trichothecene vomitoxin (VT or deoxynivalenol) may involve dysregulation of cytokine production at the T cell level. EL4.IL-2 (EL-4), a cloned thymoma that produces interleukins (IL)-2, 4, 5, and 6, was used as a T cell model to investigate the in vitro effects of VT on interleukin production and gene expression.

In vitro effects of vomitoxin (deoxynivalenol) on T-cell interleukin production and IgA secretion.

The exposure of lymphocyte cultures to vomitoxin was used to determine possible mechanisms by which this naturally occurring toxin induces serum immunoglobulin A (IgA) elevation and IgA nephropathy in the mouse. Vomitoxin exposure within the range of 10 to 1000 ng/ml inhibited DNA synthesis, protein synthesis as well as IgA, IgG and IgM production in lymphocyte cultures prepared from the Peyer's patch (PP) and spleen. When purified B cells were cultured in the presence of vomitoxin, inhibition of IgA, IgG and IgM production was similarly observed.

Polyspecific and autoreactive IgA secreted by hybridomas derived from Peyer's patches of vomitoxin-fed mice: characterization and possible pathogenic role in IgA nephropathy.

A total of 122 immunoglobulin (Ig)A-producing hybridoma clones were isolated from the Peyer's patches of vomitoxin-fed BALB/c mice and the resultant antibodies were characterized for their antigenic specificity and pathogenic potential. When reactivity was tested against a panel consisting of DNA, sphingomyelin, thyroglobulin, collagen, casein, cardiolipin and bovine serum albumin conjugates of phosphorylcholine, inulin and trinitrophenol that were representative of self and non-self antigens, approximately 95% of the monoclonal IgAs bound to at least one of the panel antigens and 80% bound to more than one antigen. The polyspecificity of some of the monoclonal IgAs was further suggested by demonstrating the capacity of one antigen to inhibit binding of monoclonal IgA to another antigen.

Polyclonal autoreactive IgA increase and mesangial deposition during vomitoxin-induced IgA nephropathy in the BALB/c mouse.

To establish the relationship between autoreactive antibodies and vomitoxin-induced immunoglobulin A (IgA) nephropathy, the effects of dietary vomitoxin exposure on the antigen specificity of serum IgA, IgA-producing cells and accumulated mesangial IgA in BALB/c mice were assessed. Exposure to dietary vomitoxin for 8 wk caused a significant increase in total serum IgA. There was a concurrent significant increase in serum IgA specific for trinitrophenol (TNP), phosphorylcholine, cardiolipin and sphingomyelin compared with controls, suggesting an elevation of autoreactive IgA.

Simultaneous screening of fumonisin B1, aflatoxin B1, and zearalenone by line immunoblot: a computer-assisted multianalyte assay system.

A line immunoblot assay was developed for the simultaneous screening of fumonisin B1 (FB1), aflatoxin B1 (AFB1), and zearalenone (ZEA). Monoclonal antibodies for each of these toxins were immobilized as multiple lines on nitrocellulose membrane strips and sectored into hydrophobic compartments to minimize use of reagents. A modified enzyme-linked immunosorbent assay was conducted whereby free mycotoxins and horseradish peroxidase-labeled mycotoxins competed for binding to the nitrocellulose-bound antibodies.

Comparative Assessment of Fumonisin in Grain-Based Foods by ELISA, GC-MS, and HPLC.

Seventy-one (71) food samples were analyzed for the mycotoxin fumonisin by a monoclonal antibody based competitive enzyme-linked immunosorbent assay (ELISA). Fumonisins were detected primarily in corn-based products with 7/12, 2/2 and 1/3 and 1/7 yellow cornmeal, blue cornmeal, corn muffin mix, and mixed grain cereal samples yielding positive results, respectively. When the positive samples and randomly selected negative samples were assessed by other methods, correlations (r values) between ELISA and gas chromatography-mass spectrometry (GC-MS), ELISA and high-pressure liquid chromatography (HPLC) and GC-MS and HPLC were 0.

Progressive serum IgE elevation in the B6C3F1 mouse following withdrawal of dietary vomitoxin (deoxynivalenol).

Vomitoxin (deoxynivalenol) is a fungal toxin that induces serum IgA hyperelevation, IgA autoantibodies, mesangial IgA deposition in mice upon dietary exposure. The capacity of dietary vomitoxin to similarly alter serum IgE was assessed in female B6C3F1 mice. Ingestion of 25 ppm vomitoxin in AIN-76A semipurified diet resulted in 2.

Production of polyclonal antibodies to the trichothecene mycotoxin 4,15-diacetylnivalenol with the carrier-adjuvant cholera toxin.

The trichothecene mycotoxin 4,15-diacetylnivalenol (DNIV) was conjugated to cholera toxin (DNIV-CT) for use as an immunogen and as an adjuvant for specific antibody production. Repeated intravenous injection of 7.5 micrograms of the conjugate was effective at generating specific antibodies to DNIV in rabbits as determined by enzyme-linked immunosorbent assay (ELISA).

Lactate Dehydrogenase as Safe Endpoint Cooking Indicator in Poultry Breast Rolls: Development of Monoclonal Antibodies and Application to Sandwich Enzyme-Linked Immunosorbent Assay (ELISA).

A sandwich enzyme-linked immunosorbent assay (ELISA) was developed to detect lactate dehydrogenase (LDH) as a marker protein for verifying endpoint cooking of uncured poultry products. Monoclonal antibodies were prepared against chicken muscle LDH and used with rabbit polyclonal antibodies developed against turkey or chicken muscle LDH for capture and detection in the assay, respectively. Minimum assay detection limits for turkey and chicken muscle LDH were 1 ng/ml.

In vitro vomitoxin exposure alters IgA and IgM secretion by CH12LX B cells. Relationship to proliferation and macromolecular synthesis.

The CH12LX cell line was used as a clonal model to assess the direct effects of vomitoxin on IgM and IgA secretion in B cells. When vomitoxin was included in LPS-driven CH12LX B cell cultures, it had multiple effects on Ig secretion. Whereas vomitoxin doses of 115 and 120 ng/ml caused 50% inhibition (ID50) of IgA and IgM production, respectively, toxin concentrations in the 5 to 50 ng/ml range slightly stimulated IgA production.

Persistent dysregulation of IgA production and IgA nephropathy in the B6C3F1 mouse following withdrawal of dietary vomitoxin (deoxynivalenol).

To assess whether vomitoxin-induced dysregulation of IgA production and IgA nephropathy are reversible, relevant immunologic parameters were compared among experimental groups of B6C3F1 mice that were fed: (1) 25 ppm vomitoxin in AIN-76A semipurified diet for 24 weeks (treatment group), (2) 25 ppm vomitoxin for 8 weeks and then control diet for 16 weeks (withdrawal group), and (3) control diet for 24 weeks (control group). Levels of serum IgA and microhematuria index in the treatment group were elevated after 4 to 8 weeks and continued to increase with further vomitoxin exposure. IgA immune complexes and mesangial IgA deposition, as quantitated by interactive laser cytometer image analysis, were also increased with toxin exposure at Weeks 8, 16, and 24, whereas IgM, IgG, and complement component C3 deposition were unaffected or depressed.

Vomitoxin-induced dysregulation of serum IgA, IgM and IgG reactive with gut bacterial and self antigens.

The effect of dietary vomitoxin exposure on immunoglobulins that react with naturally occurring gut bacterial and self antigens was assessed in the B6C3F1 mouse. Ingestion of 25 ppm vomitoxin for 4 and 8 wk resulted in significantly elevated total IgA but depressed total IgG and IgM in serum when compared with control mice fed semi-purified diet only. IgA specific for phosphorylcholine (PC) and inulin (haptens associated with intestinal bacteria) increased significantly in mice fed vomitoxin whereas IgM with the identical specificity decreased.

Generation of antibodies reactive with fumonisins B1, B2, and B3 by using cholera toxin as the carrier-adjuvant.

Murine polyclonal antibodies reactive with fumonisins B1, B2, and B3 were produced after a novel immunization procedure with cholera toxin as both a hapten carrier protein and adjuvant. Immunization of mice with two 7.5-micrograms doses of fumonisin B1-cholera toxin conjugate without adjuvant resulted in the production of fumonisin B1-specific antibodies in all mice within 15 days when intraperitoneal, subcutaneous, and intravenous routes were used.

Natural occurrence of zearalenone in rice and soybean produced in Korea.

88 rice and 75 soybean samples were collected from 8 provinces of Korea from March through September in 1988. The Fusarium mycotoxins, zearalenone was analyzed by direct competitive enzyme linked immunosorbent assay. 10.