concanavalin-a and 3-acetyldeoxynivalenol

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. At day 2, all trichothecenes were found to have inhibited production of IL-2, IL-4, and IL-5. However, at day 7, supernatant IL-2 was significantly increased (2-5.5-fold) in cultures containing VT, NIV, 3-ADON, and 15-ADON at 250, 250, 2500, and 1000 ng/ml doses, respectively, when compared to control Con A-stimulated cultures; significant increases in IL-2 were not observed with T-2 and Ver-A. Similarly, at day 7, IL-4 and IL-5 were significantly increased in the presence of VT (100 ng/ml), NIV (100 ng/ml), 3-ADON (1000 ng/ml), 15-ADON (500 ng/ml), T-2 (1 ng/ml), and Ver A (50 pg/ml, only IL-5) when compared to control cultures. IL production was inhibited at trichothecene concentrations exceeding the aforementioned optima. When total RNA of 2-day cultures was assessed by reverse transcriptase polymerase chain reaction (RT-PCR) in conjunction with Southern analysis, IL-2 mRNA was also found to be superinduced by VT (50 and 100 ng/ml), NIV (50, 100 and 250 ng/ml), 3-ADON (1500 ng/ml), 15-ADON (100 ng/ml), T-2 (0.5 ng/ml) and Ver A (25, 50 and 100 pg/ml); IL-4 mRNA by VT (50 ng/ml), NIV (50 ng/ml), and Ver A (25, 50 and 100 pg/ml); IL-5 mRNA by VT (50 ng/ml); and IL-6 mRNA by 15-ADON (100 ng/ml) and Ver A (50 pg/ml). As the trichothecene concentration increased from these levels, inhibition of mRNA transcript levels was also observed for many of the interleukins. Taken together, the results suggest that trichothecenes as a group can either inhibit or superinduce both IL secretion and mRNA levels in CD4+ T-cells. Superinduction exhibited a rank order of macrocyclic > type A > type B trichothecenes and was dependent on acylation of the trichothecene nucleus.

Topics: Acylation; Animals; Antibiotics, Antineoplastic; Blotting, Southern; CD4-Positive T-Lymphocytes; Cells, Cultured; Concanavalin A; Cytokines; Enzyme-Linked Immunosorbent Assay; Gene Expression Regulation; Interleukin-2; Interleukin-4; Interleukin-5; Mice; RNA, Messenger; Structure-Activity Relationship; T-2 Toxin; Transcription, Genetic; Trichothecenes

The effects of 3-acetyldeoxynivalenol (3-AcDON) on mitogen-induced lymphocyte proliferation and antibody production were studied in male CD-1 mice exposed to 0, 2.5, 5 or 10 ppm 3-AcDON in the diet for 35 days. Mitogen-induced lymphocyte proliferation and T-cell-independent antibody responses to dinitrophenyl-ficoll or Escherichia coli were not altered by dietary exposure to 3-AcDON. The T-cell-dependent antibody response to sheep red blood cells was increased in the group fed 10 ppm 3-AcDON. In vitro, 3-AcDON inhibited lymphocyte proliferation in a dose-dependent manner. Inhibition was observed when the toxin was present during the first 8 hr in phytohaemagglutinin-stimulated cultures and during the first 24 hr in lipopolysaccharide-stimulated cultures. This suggests that 3-AcDON blocks an early step in lymphocyte activation. This inhibition was not restored by thiol reducing agents (dithiothreitol, L-cysteine or 2-mercaptoethanol). Similarly, the addition of lymphokines, including interleukin-1 or interleukin-2, did not alter the inhibitory effects of 3-AcDON. These results suggest that the in vitro effects of 3-AcDON may not reflect its in vivo immunotoxicity. However, 3-AcDON may serve as a chemical probe for examining the activation process of lymphocyte proliferation.

Topics: Animals; Antibody Formation; Concanavalin A; Lipopolysaccharides; Lymphocyte Activation; Lymphokines; Male; Mice; Sesquiterpenes; Spleen; Trichothecenes

The effects of 3-acetyl-deoxynivalenol (3-AcDON) on the in vitro mitogen responses and the antibody producing ability of human peripheral blood lymphocytes were evaluated. 3-AcDON inhibited the proliferative response to pokeweed mitogen and concanavalin A at a lower concentration (100 ng/ml) as compared to phytohemagglutinin (200 ng/ml). The antibody producing ability was inhibited by 3-AcDON concentrations of greater than or equal to 200 ng/ml. Higher concentrations of 3-AcDON (greater than or equal to 300 ng/ml) produced severe suppression of plaque forming cell response in vitro and reduced the total yield of lymphocytes without altering cell viability. The results of this study indicate that 3-AcDON produces immunosuppressive effects in a dose dependent manner in vitro.

Topics: Adult; Antibody Formation; Concanavalin A; Humans; Immunosuppressive Agents; In Vitro Techniques; Lymphocyte Activation; Lymphocytes; Phytohemagglutinins; Pokeweed Mitogens; Sesquiterpenes; Trichothecenes