Interactions of foot-and-mouth disease virus with cells in organised lymphoid tissue influence innate and adaptive immune responses
Juleff, Nicholas Dylan
Foot-and-mouth disease virus (FMDV) is one of the most contagious viruses of animals and is recognised as the most important constraint to international trade in animals and animal products. Two fundamental problems remain to be understood before more effective control measures can be put in place. These problems are the FMDV „carrier state‟ and the short duration of immunity after vaccination which contrasts with prolonged immunity after natural infection. The aim of this thesis was to study the interaction between FDMV and cells in lymphoid tissue in the natural bovine host, in order to improve our understanding of the protective immune response. Using laser capture microdissection in combination with quantitative real-time reverse transcription polymerase chain reaction, immunohistochemical analysis and corroborated by in situ hybridization, it is shown that FMDV locates rapidly to, and is maintained in, the light zone of germinal centres following primary infection of naïve cattle. Maintenance of non-replicating FMDV in these sites may represent a source of persisting infectious virus and also contribute to the generation of long-lasting antibody responses against neutralising epitopes of the virus. The role of T-lymphocyte subsets in recovery from FMDV infection in calves was investigated by administering subset-specific mouse monoclonal antibodies. Depletion of circulating CD4+ or WC1+ γδ T cells was achieved for a period extending from before challenge to after resolution of viraemia and peak clinical signs, whereas CD8+ cell depletion was only partial. Depletion of CD4+ cells was also confirmed by analysis of lymph node biopsies 5 days post-challenge. Depletion with anti-WC1 and anti-CD8 antibodies had no effect on the kinetics of infection, clinical signs and immune responses following FMDV infection. Three of the four CD4+ T-cell-depleted calves failed to generate an antibody response to the non-structural polyprotein 3ABC, but generated a neutralising antibody response similar to that in the controls, including rapid isotype switching to IgG antibody. These data suggest that antibody responses to sites on the surface of the virus capsid are T cell-independent whereas those directed against the non-structural proteins are T cell-dependent. CD4 depletion was found to substantially inhibit antibody responses to the G-H peptide loop VP1135-156 on the viral capsid, indicating that responses to this particular site, which has a more mobile structure than other neutralising sites on the virus capsid, are T cell-dependent. Depletion of CD4+ T cells had no adverse effect on the magnitude or duration of clinical signs or clearance of virus from the circulation. In conclusion, CD4+ T-cell-independent antibody responses play a major role in the resolution of primary infection with FMDV in cattle.