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SUPPLEMENT. FOOD SAFETY CONCERNS OF VEROTOXIN-PRODUCING ESCHERICHIA COLI

Verotoxin 1 from Escherichia coli Affects Gb₃/CD77⁺ Bovine Lymphocytes Independent of Interleukin-2, Tumor Necrosis Factor-, and Interferon-

Christian Menge^*,1, Ivonne Stamm^*, Maike Blessenohl^*, Lothar H. Wieler^*, and Georg Baljer^*

^* Institut für Hygiene und Infektionskrankheiten der Tiere, Justus-Liebig-Universität, D-35392 Giessen; and
Institut für Mikrobiologie und Tierseuchen, Freie Universität Berlin, D-10115 Berlin, Germany

Abstract

Verotoxin (VT)-induced immunomodulation has been implicated in the ability of VT-producing Escherichia coli (VTEC) to cause persistent infections in cattle. VT1, also referred to as Shiga toxin 1, is a potent cytotoxin that modulates cytokine secretions and functions. This prompted the current investigation to examine whether the inhibiting effect of VT1 on bovine lymphocytes correlates with the expression of the cellular VT1 receptor Gb₃/CD77 or is mediated instead via perturbation of cytokine secretion. Using blood mononuclear cells stimulated by mitogens as a model, VT1 significantly blocked lymphoblast transformation and proliferation in the BoCD8⁺ T cell and BoCD21⁺ B cell population. In contrast, VT1 dramatically reduced the number of viable Gb₃/CD77⁺ blast cells within all subpopulations identified (BoCD2⁺, BoCD4⁺, BoCD8⁺, WC1⁺ [i.e., T cells] BoCD21⁺, and BoCD25⁺). Similar effects of VT1 were observed when the culture medium was supplemented with selected cytokines: tumor necrosis factor--sensitizing endothelial cells against VT1, interferon- (IFN-) as bovine IFN- receptors are partially homologous to the B-subunit of VT1, and interleukin-2 that is critical for lymphocyte proliferation in vitro. The addition of these cytokines was neither able to mimic nor to overcome the effects of VT1. Therefore, it is concluded that VT1 directly acts on bovine lymphocytes rather than inducing a cytokine-mediated effect. VT1 considerably affects all main bovine lymphocyte subpopulations, implicating that the immune system is a predominant target for VT1 in cattle.

Key Words: verotoxins • immunity • cattle • Shiga toxin • Escherichia coli • food safety

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