Date Published: May 14, 2008
Publisher: Public Library of Science
Author(s): John K. Thuita, John M. Kagira, David Mwangangi, Enock Matovu, C. M. R. Turner, Daniel Masiga, Serap Aksoy
Abstract: We have investigated the pathogenicity of tsetse (Glossina pallidipes)-transmitted cloned strains of Trypanosoma brucei rhodesiense in vervet monkeys. Tsetse flies were confirmed to have mature trypanosome infections by xenodiagnosis, after which nine monkeys were infected via the bite of a single infected fly. Chancres developed in five of the nine (55.6%) monkeys within 4 to 8 days post infection (dpi). All nine individuals were successfully infected, with a median pre-patent period of 4 (range = 4–10) days, indicating that trypanosomes migrated from the site of fly bite to the systemic circulation rapidly and independently of the development of the chancre. The time lag to detection of parasites in cerebrospinal fluid (CSF) was a median 16 (range = 8–40) days, marking the onset of central nervous system (CNS, late) stage disease. Subsequently, CSF white cell numbers increased above the pre-infection median count of 2 (range = 0–9) cells/µl, with a positive linear association between their numbers and that of CSF trypanosomes. Haematological changes showed that the monkeys experienced an early microcytic-hypochromic anaemia and severe progressive thrombocytopaenia. Despite a 3-fold increase in granulocyte numbers by 4 dpi, leucopaenia occurred early (8 dpi) in the monkey infection, determined mainly by reductions in lymphocyte numbers. Terminally, leucocytosis was observed in three of nine (33%) individuals. The duration of infection was a median of 68 (range = 22–120) days. Strain and individual differences were observed in the severity of the clinical and clinical pathology findings, with two strains (KETRI 3741 and 3801) producing a more acute disease than the other two (KETRI 3804 and 3928). The study shows that the fly-transmitted model accurately mimics the human disease and is therefore a suitable gateway to understanding human African trypanosomiasis (HAT; sleeping sickness).
Partial Text: In human African trypanosomiasis (HAT), the use of animal models has contributed enormously to what is currently known about the relationships between disease duration, parasite invasion of different body systems and the potential of resultant host clinical and biological changes as diagnostic and disease staging markers. Several host-parasite model systems have been developed, based on infection of various hosts with the livestock pathogen Trypanosoma brucei brucei and to a lesser extent the human pathogens T. b. rhodesiense and T. b. gambiense. Characterisation of these HAT models shows that the disease occurs in two stages irrespective of host: an early haemo-lymphatic trypanosome proliferation, and a late central nervous system (CNS) infection, indicating that the basic pattern is similar to the disease in humans. This is evidenced by demonstration of trypanosomes, first in the haemo-lymphatic system and later in the CNS of the mouse model with subsequent cerebral pathology ,. Models based on larger mammals such as the chimpanzee T. b. rhodesiense model , the vervet monkey T. b. rhodesiense model  and the sheep T. b. brucei model , also follow a similar two-stage disease pattern. These, unlike rodents, allow collection of cerebrospinal fluid (CSF) that has been used to demonstrate elevation of white cell counts and total protein levels as indicators of CNS stage disease .
In this study, infection of vervet monkeys was initiated by the bite of a single infected tsetse fly. To our knowledge, this represents the first time single fly transmission of T. b. rhodesiense clones in vervet monkeys (or any other primate model) has been achieved, hence establishing a model that more accurately mimics the transmission of sleeping sickness as it occurs in humans. Information on natural HAT relies on data provided in case reports – and sometimes re-analysis of retrospective clinical, epidemiological and pathology data –. Such data are naturally limited on the questions of disease onset and duration as there are only a small number of case reports in which the patient could accurately remember the exact time of being bitten by a tsetse fly ,. In any case this is not readily accessible information for inhabitants of endemic areas, where tsetse fly challenge is continuous. This study has allowed us to generate information on the pathogenesis of HAT in a manner that more accurately mimics human disease, and facilitates documentation of data from precise sampling points during the course of a tsetse transmitted infection.