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Title: Cellular immune responses of cattle to Escherichia coli O157:H7
Author: Corbishley, Alexander
ISNI:       0000 0004 5991 3998
Awarding Body: University of Edinburgh
Current Institution: University of Edinburgh
Date of Award: 2015
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Enterohaemorrhagic Escherichia coli O157:H7 causes haemorrhagic diarrhoea and potentially fatal renal failure in humans. Ruminants are considered to be the primary reservoir for human infection. Vaccines that reduce shedding in cattle are only partially protective and their underlying protective mechanisms are unknown. Studies investigating the response of cattle to colonisation generally focus on humoral immunity, leaving the role of cellular immunity unclear. To inform future vaccine development, the cellular immune response of cattle during EHEC O157:H7 colonisation was examined. Calves were challenged with either a phage type (PT) 21/28 strain possessing the Shiga toxin (Stx) 2a and Stx2c genes or a PT32 strain possessing the Stx2c gene only. T-helper cell associated transcripts at the terminal rectum were analysed by reverse transcriptase quantitative PCR (RT-qPCR). Induction of interferon (IFN)γ and T-bet was observed, with peak expression of both genes at 7 days in PT32 challenged calves, whilst up regulation was delayed, peaking at 21 days in PT21/28 challenged calves. Cells isolated from gastro-intestinal lymph nodes demonstrated antigen-specific proliferation and IFNγ release in response to type III secreted proteins (T3SPs); however responsiveness was suppressed in cells isolated from PT32 challenged calves. Lymph node cells showed increased expression of the proliferation marker Ki67 in CD4+ T cells from PT21/28, NK cells from PT32 and CD8+ and γδ T cells from both PT21/28 and PT32 challenged calves following ex vivo stimulation with T3SPs. Epitope mapping of rectal lymph node CD4+ T cell responses to 16 EHEC O157:H7 proteins, identified 20 CD4+ T cell epitopes specific to E. coli. The highly conserved N-terminal region of Intimin, including the signal peptide, was consistently recognised by mucosal CD4+ T cell populations from multiple animals of different major histocompatibility complex (MHC) class II haplotypes. Studies investigating the impact of secreted bacterial proteins on bovine peripheral blood mononuclear cells (PBMC) identified the ability of these proteins to cleave the surface molecule CD8 and that this phenotype was dependent on the ler virulence regulator but not the type III secretory system (T3SS) machinery. This effect was also observed in murine and ovine, but not human lymphocytes. Preliminary in vitro experiments suggest that this activity may reduce the efficiency of CD8+ T cell killing. This study demonstrates that cattle mount cellular immune responses during colonisation with EHEC O157:H7, the temporality of which is strain dependent, with further evidence of strain-specific immunomodulation.
Supervisor: Gally, David ; Mcneilly, Tom Sponsor: Biotechnology and Biological Sciences Research Council (BBSRC)
Qualification Name: Thesis (Ph.D.) Qualification Level: Doctoral
EThOS ID:  DOI: Not available
Keywords: Escherichia coli ; e-coli ; O157 ; cattle ; vaccine ; cellular immunity