Bovine immune responses to cattle tick infestation

The cattle tick, Rhipicephalus Boophilus microplus (formerly Boophilus microplus), is the most significant parasite of cattle in Australia and threatens the improvement of cattle production in tropical and subtropical countries worldwide. Some cattle breeds of mainly Bos indicus origin develop a strong resistance to infestation with R. B. microplus, while other breeds of mainly Bos taurus origin will succumb to anaemia and ‘tick worry’ in tick-infested pastures. Resistance to cattle tick infestation is primarily manifest against the larval stage and results in the immature tick failing to make a successful attachment and obtain a meal. It is widely accepted that resistance to tick infestation in cattle is immunologically mediated and involves both cell-mediated and humoral immune responses. Previous research has demonstrated a role for the hypersensitivity response in the rejection of larvae from resistant B. taurus cattle. The immune response to R. B. microplus infestation in B. indicus and B. indicus × B. taurus cattle has not been studied in great detail and this thesis aimed to describe those responses of highly tick-resistant cattle that differ from those of tick-susceptible cattle. A pilot trial was conducted using six tick-resistant Brahman heifers (B. indicus) and six tick-susceptible Holstein-Friesian heifers (B. taurus). The animals were artificially infested for several weeks and blood samples obtained weekly for three weeks during the height of infestation, and on one occasion, skin biopsies obtained. It was found that significant differences existed between the two breeds with respect to the percentage of cellular subsets comprising the peripheral blood mononuclear cell (PBMC) population, cytokine expression by peripheral blood leukocytes (PBL), and levels of tick-specific immunoglobulin G1 (IgG1) antibodies measured in the peripheral circulation. These results in combination with whole genome expression analysis of circulating PBL suggested that the B. indicus cattle had developed a stabilised T cell-mediated response to tick infestation evidenced by their cellular profile and leukocyte cytokine spectrum. The B. taurus cattle demonstrated cellular and gene expression profiles consistent with a sustained innate, inflammatory response to infestation, although high tick-specific IgG1 levels suggested that these animals had also developed a T cell response to infestation. Gene expression analyses using quantitative PCR (qPCR) and whole genome expression platforms indicated that the B. taurus cattle elicited an innate inflammatory response in the skin at the site of larval attachment. This was supported by histological examination of the tick-attachment sites. Conversely, the B. indicus cattle appeared to limit the inflammatory response and instead exhibited increased expression of genes involved in wound healing and the structural integrity of the skin. A second trial was conducted using a group of tick-naïve Santa-Gertrudis heifers to eliminate any innate breed differences that may have confounded the results obtained in the pilot trial. Thirty-five animals were obtained from a tick-free region of Australia; thirty animals were artificially infested weekly for thirteen weeks while five animals remained at a tick-free quarantine property to serve as a control group. Following thirteen weeks of tick infestation the animals in this trial exhibited divergent tick-resistance phenotypes and those carrying the lowest and highest numbers of ticks were classified into tick-resistant (n = 6 animals) and tick-susceptible (n = 6 animals) groups, respectively. The larger number of animals in this study and repeated measurements over the course of the trial enabled a detailed analysis of the effect of tick infestation across all animals, and also within the groups identified as highly resistant or highly susceptible. A cell-mediated response to tick-infestation was detected in all tick-infested animals by twenty-one days after the initial infestation, a response which waned towards the end of the trial when the antibody response became more dominant. Similar to the pilot trial, the tick-susceptible animals developed significantly higher levels of tick-specific IgG1 antibodies compared with the tick-resistant group. Some differences were observed between tick-resistant and tick-susceptible animals with respect to the percentage of cellular subsets comprising the PBMC population. Skin biopsies were taken from these animals prior to initial infestation and then at twenty-one days and ninety-one days after the initial infestation and whole genome expression profiles were produced and analysed. At twenty-one days post infestation both tick-resistant and tick-susceptible animals demonstrated an upregulation of genes involved in leukocyte migration and complement activation, suggestive of an inflammatory response. While this response persisted in the tick-susceptible group at ninety-one days post-infestation, it appeared to lessen in the tick-resistant group. A dominant interleukin-8 (IL-8) response was observed in the skin of the tick-susceptible animals at both time points post-infestation that was not observed in the tick-resistant animals. This thesis demonstrates that tick-susceptible animals respond to infestation with R. B. microplus with an apparently non-protective antibody response, and that tick-resistant animals demonstrate different structural responses in the skin at the tick-attachment site compared with the intense inflammatory response observed in tick-susceptible animals. The results presented here suggest that the extreme susceptibility of pure B. taurus and some B. taurus × B. indicus individuals may be due to their heightened ability to recognise and respond to tick antigens introduced into the host via tick saliva during the blood feeding process. These animals’ increased ability to recognise and respond to these proteins may act to set up a chronic state of inflammation that is beneficial to the tick through increased capillary permeability providing a suitable environment for tick feeding and survival