Flow Cytometric Analysis of the Cytotoxic T-Cell Recall Response to Theileria parva in Cattle Following Vaccination by the Infection and Treatment Method

The apicomplexan hemoparasite, Theileria parva, causes East Coast fever (ECF), a frequently fatal disease of African cattle. Vaccine development has been impeded by incomplete understanding of protective immunity following natural exposure or the infection and treatment method (ITM) of immunization. This is attributable to a paucity of methods to characterize the memory T-cell repertoire following infection. To overcome this impediment, assays developed to study the immune response to other intracellular pathogens were adapted for use in studies with T. parva to enable definition of the phenotype and function of effector T cells in T. parva-immune cattle, facilitating vaccine development. As reported herein, stimulation of peripheral blood mononuclear cells (PBMC) from ITM-immunized steers with irradiated, autologous, T. parva-infected cell lines elicited a proliferative recall response comprised of CD45R0+/CCR7− CD4+ and CD8+ T cells. Subsequent co-incubation of stimulated cultures with infected cells demonstrated the presence of cytotoxic T cells (CTLs) with the ability to kill infected cells. Comparison of CTL activity in cultures depleted of CD4+ or CD8+ T cells demonstrated CTL activity was primarily attributed to CD8+ T cells. Importantly, stimulation of PBMC from vaccinated steers always elicited proliferation of CD4+ and CD8+ T cells. This was the first important observation obtained from the use of the assay described herein.

Trends in the control of theileriosis in sub-Saharan Africa : tick-borne diseases

The declining efficacy of acaricide treatment as a means of reducing the prevalence of Theileria parva infections in sub-Saharan Africa has intensified efforts to achieve control through immunization of susceptible cattle. The infection and treatment method of immunization has enjoyed a resurgence with the availability of more effective cold chain facilities, although concerns remain regarding the possibility of vaccine strains spreading in local tick populations. In addition, an in-depth understanding of protective mechanisms deployed by immune cattle and the antigens targeted by them has led to substantial progress in the development of candidate subunit vaccines against both sporozoite and schizont stages of the parasite. The likely success of these vaccines, as well as infection and treatment immunization, will ultimately depend on the extent to which they disturb the endemic status of the parasite. These issues are discussed in the light of recent information on the genotypic diversity of T. parva in the field and the extent to which this is compromised by the immune response.

Comparison of manual and homogenizer methods for preparation of tick-derived stabilates of Theileria parva: equivalence testing using an in vitro titration model

Theileria parva sporozoite stabilates are used in the infection and treatment method of immunization, a widely accepted control option for East Coast fever in cattle. T. parva sporozoites are extracted from infected adult Rhipicephalus appendiculatus ticks either manually, using a pestle and a mortar, or by use of an electric homogenizer. A comparison of the two methods as a function of stabilate infectivity has never been documented. This study was designed to provide a quantitative comparison of stabilates produced by the two methods. The approach was to prepare batches of stabilate by both methods and then subject them to in vitro titration. Equivalence testing was then performed on the average effective doses (ED). The ratio of infective sporozoites yielded by the two methods was found to be 1·14 in favour of the manually ground stabilate with an upper limit of the 95% confidence interval equal to 1·3. We conclude that the choice of method rests more on costs, available infrastructure and standardization than on which method produces a richer sporozoite stabilate.