Publications

Serum samples collected monthly over a 34-month period from cattle, sheep and goats in the Greater Accra Region of Ghana were tested for antibodies to Ehrlichia (previously Cowdria) ruminantium, the causative agent of heartwater, by polyclonal competitive ELISA (PC-ELISA). Maternal antibodies, detected in about half of animals followed from under 1 month old, declined to negative levels within 2-4 months. Amblyomma variegatum tick vectors were present on livestock in rural areas throughout the year, and first seroconversion occurred at any age, although the majority of calves seroconverted between 1 and 10 months old, sheep by 11 months, and goats by 7 months. All the cattle in the study became seropositive by 20 months of age, except one animal which subsequently died of heartwater. Following seroconversion, 25% of bovine sera tested negative in the PC-ELISA. Just over half the sheep in the survey seroconverted before or during the study period; following seroconversion, less than 3% of ovine sera became PC-ELISA negative. About a quarter of the goats seroconverted, and 34% of their post-seroconversion sera tested negative in the PC-ELISA. Overall, the serology indicated that virtually all cattle on the survey farms were exposed to E. ruminantium without suffering disease, but that a substantial proportion of sheep and goats escaped exposure and thus formed a susceptible population. E. ruminantium was detected in brains of 14, 36 and 4% of cattle, sheep and goats submitted for post mortem at the Accra Veterinary Laboratory, indicating that sheep were most at risk from heartwater disease.

Hemophagocytic lymphohistiocytosis (HLH) and Langerhans cell histiocytosis (LCH) are members of a group of rare heterogenous disorders, the histiocytoses, characterized by uncontrolled accumulation of pleomorphic infiltrates of leukocytes. The etiology of these diseases is mainly unknown. CD45 is a hemopoietic cell specific tyrosine phosphatase essential for antigen receptor mediated signaling in lymphocytes and different patterns of CD45 splicing are associated with distinct functions. Recently a polymorphism (C77G) in exon 4 of CD45 causing abnormal CD45 splicing and a point mutation affecting CD45 dimerization were implicated in multiple sclerosis in humans and lymphoproliferation and autoimmunity in mice respectively. Here we show that two patients with HLH exhibited abnormal CD45 splicing caused by the C77G variant allele, while a further 21 HLH patients have normal CD45. We have also examined 62 LCH patients and found three to have the C77G mutation. Peripheral blood thymus-derived (T) CD8(+) cells from normal individuals carrying the C77G mutation show a significant decrease in the proportion of cells expressing L-selectin and increased frequency of cells with LFA-1(hi) expression. It remains to be established whether C77G is a contributing factor in these histiocytic disorders.

The aim of this study was to determine the expression of three proinflammatory cytokines by pulmonary macrophages of sheep in paraffin wax-embedded tissue. Samples of lung from seven healthy sheep were fixed by immersion in either 10% neutral buffered formalin, acetic formalin, paraformaldehyde-lysine-periodate or Bouin's solution and processed for structural and immunohistochemical studies. The expression of interleukin (IL)-1alpha-, IL-6 and tumour necrosis factor (TNF)-alpha by pulmonary intravascular macrophages (PIMs) and alveolar macrophages (AMs) was detected by the avidin-biotin-peroxidase (ABC) technique. Bouin's solution proved to be the most suitable fixative and Tween 20(R) the most effective pretreatment for increasing permeability. Constitutive expression of IL-1alpha, IL-6 and TNF-alpha by both macrophage populations was detected. The number of PIMs expressing IL-1alpha (the predominant cytokine in ovine lung) was higher than that of AMs, while the expression of IL-6 was greater in AMs. No differences between PIMS and AMs were found in respect of TNF-alpha expression. The evaluation of cytokine expression represents a valuable tool for studying the pathogenesis of disease in the ovine lung.

The interaction of mycobacteria with antigen presenting cells is a key feature in the pathogenesis of tuberculosis and the outcome of this interaction is pivotal in determining whether immunity or disease ensues. Human and mouse macrophages and dendritic cells (DC) have been shown to become infected with mycobacteria and to produce a response to infection that reflects their suggested role in immunity. Thus, macrophages elicit anti-microbial mechanisms for elimination of mycobacteria and DC up-regulate expression of molecules that aid their stimulation of T lymphocytes. We have examined the effects of infection with the avirulent strain Mycobacterium bovis BCG and with virulent M. bovis on bovine antigen presenting cells. Differences in the intracellular survival of bacteria within DC and macrophages were observed with higher numbers of bacteria maintained within DC following infection compared to macrophages. BCG was killed more effectively than M. bovis. Alterations in the expression of cell surface molecules involved in antigen presentation and the stimulation of T cells, including MHC II and CD40, were observed following infection of bovine antigen presenting cells. In addition infected DC secreted IL-12, TNF? and IL-10 whereas macrophages produced TNF?, IL-10 and little IL-12. Generally responses were more marked when virulent M. bovis was used compared to BCG. These studies indicate that infection of bovine antigen presenting cells by mycobacterial species results in the induction of both innate and adaptive immune responses that are critical for the outcome of infection.