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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 Abstract
                                    Publication Type: Abstract Only
                                    Publication Acceptance Date: 11/6/2014
                                    Publication  Date: 1/23/2015
                                    Citation: Maggioli, M.F., Palmer, M.V., Vordermeier, H.M., Whelan, A.O., Waters, W.R. 2015. Polyfunctional cytokine responses by central memory CD4+T cells in response to bovine tuberculosis [abstract]. Keystone Symposium. Abstract No. 33.Interpretive Summary:
                                Technical Abstract: CD4 T cells are crucial in immunity to tuberculosis (TB). 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. Mycobacterium bovis infection of cattle elicits ex vivo polyfunctional T cell responses. Long-term (i.e., 14 days) cultured PBMCs are rich in T cell central memory cells (Tcm) in both humans and cattle. The objective of the study was to assess polyfunctional T cell responses by Tcm in cattle (n = 6) in response to aerosol M. bovis infection. For long-term culture, PBMCs from infected cattle were stimulated with M. bovis purified protein derivative, rESAT-6:CFP-10 (E:C) and a cocktail of rTb10.4 and rAg85A for 13 days with periodic addition of fresh media and rIL-2. After stimulation, the expression of CD4, CD45RO, CCR7, IL-2, IFN-gamma, TNF-alpha and cell viability were analyzed by flow cytometry. Long-term T cells responding to E:C consisted of 11% effectors (CD4+ CD45RO- CCR7-), 22% effector memory (CD4+ CD45RO+ CCR7-), and 76% central memory (CD4+ CD45RO+ CCR7+) phenotypes. In regard to the cytokine profile of responding cells, 53% were producing IFN-gamma and TNF-alpha, 42% produced all three cytokines, 13% produced TNF-alpha and IL-2, 7% produced IL-2 and IFN-gamma. Cells producing only IFN-gamma, IL-2, or TNF-alpha represented, 9%, 5% and 1%, respectively. These findings demonstrate that 14-day cultured cells (primarily Tcm cells) exhibit polyfunctional responses consisting mainly of cells co-producing IFN-gamma and TNF-alpha or IL-2, IFN-gamma and TNF-alpha.

This work was supported by Agriculture and Food Research Initiative Competitive Grant 2011-67015-30736 from the USDA.


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