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Title:   Author  item
                                            MAGGIOLI, M - Iowa State Universityitem
                                            Palmer, Mitchellitem
                                            VORDERMEIER, H - Veterinary Laboratories Agency (VLA)item
                                            WHELAN, A - Veterinary Laboratories Agency (VLA)item
                                            Waters, Wade
  Submitted to: Meeting Proceedings
                                    Publication Type: Proceedings
                                    Publication Acceptance Date: 2/17/2014
                                    Publication  Date: 5/2/2014
                                    Citation: Maggioli, M.F., Palmer, M.V., Vordermeier, H.M., Whelan, A.O., Waters, W.R. 2014. Polyfunctional CD4 T cells in the response to bovine tuberculosis. Proceedings of the American Association of Immunologists Annual Meeting. p. 64.Interpretive Summary:
                                Technical Abstract: Polyfunctional CD4 T cells simultaneously produce interferon-gamma (IFN-gamma), interleukin-2 (IL-2) and tumor necrosis factor-alpha (TNF-alpha) and play relevant roles in several chronic infections, including human TB and HIV. However, the assessment of this response in bovine infections was not feasible due to the lack of monoclonal antibodies that recognize bovine IL-2. Recently, an antibody specific for bovine IL-2 enabled the evaluation of polyfunctional T cells in cattle. The objective of the present study was to access antigen-specific polyfunctional ex vivo responses after aerosol Mycobacterium bovis infection of cattle. Peripheral blood mononuclear cells (PBMCs) were collected from infected cattle and stimulated with rESAT-6:CFP-10 (E:C), media or pokeweed (PWM) for 16 hours. After stimulation, the expression of CD4, CD45RO, CCR7, IL-2, IFN-gamma, TNF-alpha and cell viability were evaluated. The ex vivo response to E:C consisted of 63% effectors (CD4+ CD45RO- CCR7-), 32% effector memory (CD4+ CD45RO+ CCR7-), and 9% central memory (CD4+ CD45RO+ CCR7+) phenotypes. In regard to the cytokine profile, 70% of cells producing cytokines expressed both IFN-gamma and TNF-alpha, 31% expressed all three cytokines, 6% expressed both TNF-alpha and IL-2, and 3% expressed IL-2 and IFN-gamma. Cells producing only IFN-gamma, IL-2, or TNF-alpha represented, 5%, 8% and 1%, respectively. These findings demonstrate that the polyfunctional response consists mainly of effector cells co-producing IFN-gamma and TNF-alpha.


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