A kit for in vitro isolation of trypanosomes in the field: first trial with sleeping sickness patients in the Congo Republic.
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Biomedical subjects
Publications and source records attributed to D Le Ray.
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The distribution of B-(Ig+) and T-(Thy1.2+, Lyt1+, Lyt2+) lymphocyte subsets in murine lymphoid organs was analysed by immunofluorescence (FACS) on cell suspensions throughout a primary infection of C3H/He (susceptible) and CBA/Ca (subtolerant) inbred mice with metacyclic Trypanosoma brucei brucei EATRO 1125. A direct correlation was observed between (1) the level of first-peak parasitaemia, (2) its subsequent control by the host and (3) survival time. In the course of the infection, the overall population of spleen and lymph node lymphocytes was subject to polyclonal activation whilst the proportion of differentiated B and T subsets decreased accordingly. No correlation was found between modulation of the lymphocyte system and susceptibility to trypanosomiasis.
Visceral leishmaniasis (VL), also known as kala-azar, is a vector-borne disease caused by a protozoan of the Leishmania donovani complex. A phlebotomine sandfly transmits the parasite from person to person or via an animal reservoir. VL is a severe, debilitating disease, characterized by prolonged fever, splenomegaly, hypergammaglobulinaemia and pancytopenia. Patients become gradually ill over a period of a few months, and nearly always die if untreated. Case-fatality ratios are high even in treated patients. Worldwide an estimated 500,000 VL cases occur each year. This study reviews clinical, epidemiological and public health aspects of the disease and shows how critical adequate case detection is for the success of VL control. Examination of the issue of VL diagnosis with respect to the global challenges in VL control leads to the observation that a sound diagnostic-therapeutic algorithm for the health services in endemic areas is badly needed. Serological tests could be an alternative to parasitological diagnosis and the direct agglutination test (DAT) was found to fulfil many criteria for a 'field test', including cost effectiveness. Although research needs on vaccine and better drugs continue to be high on the agenda, a VL test-treatment strategy based on currently available highly sensitive serological tests, such as the DAT, should be introduced in the health services in endemic areas.
A total of 116 sera of patients suffering from South American mucocutaneous leishmaniasis were analysed using the RAST (L. braziliensis-specific) and RIST techniques. Results of total and specific IgE levels were correlated with different clinical stages of the disease. The level of total and specific IgE appeared to be higher among patients with cutaneous ulcers than among those presenting mucocutaneous lesions, i.e. espundia. Furthermore, non-treated patients presented higher concentrations of total and specific IgE than did treated ones.
A set of 38 Leishmania stocks from the Andean valleys of Peru was characterized by both Multilocus Enzyme Electrophoresis (MLEE) and Random Amplified Polymorphic DNA (RAPD). Data were analyzed in terms of taxonomy and evolutionary genetics. Synapomorphic MLEE and RAPD characters, clear-cut clustering, and strong agreement between the phylogenies inferred from either MLEE or RAPD supported the view that Leishmania (Viannia) peruviana and Leishmania (Viannia) braziliensis correspond to two closely related, but distinct monophyletic lines (clades) and can therefore be considered as "discrete typing units" (DTUs). The question whether the L. (V.) peruxviana DTU deserves species status is dependent upon the desirability of it, in terms of epidemiological and medical relevance. A previous Orthogonal Field Alternating Gel Electrophoresis (OFAGE) analysis of the same L. (V.) peruviana isolates was published by Dujardin et al. (1995b). The data from the different markers (i.e. MLEE, RAPD and OFAGE) were compared by population genetics analysis. RAPD and OFAGE provided divergent results, since RAPD showed a strong linkage disequilibrium whereas OFAGE revealed no apparent departure from panmictic expectation. MLEE showed no linkage disequilibrium. Nevertheless, contrary to OFAGE, this is most probably explainable by the limited variability revealed by this marker in L. (V.) peruviana (statistical type II error). RAPD data were consistent with the hypothesis that the present L. (V.) peruviana sample displays a basically clonal population structure with limited or no genetic exchange. Disagreement between RAPD and OFAGE can be explained either by accumulation of chromosomal rearrangements due to amplification/deletion of repeated sequences, or by pseudo-recombinational events.
Most molecular trees of trypanosomatids are based on point mutations within DNA sequences. In contrast, there are very few evolutionary studies considering DNA (re) arrangement as genetic characters. Waiting for the completion of the various parasite genome projects, first information may already be obtained from chromosome size-polymorphism, using the appropriate algorithms for data processing. Three illustrative models are presented here. First, the case of Leishmania (Viannia) braziliensis/L. (V.) peruviana is described. Thanks to a fast evolution rate (due essentially to amplification/deletion of tandemly repeated genes), molecular karyotyping seems particularly appropriate for studying recent evolutionary divergence, including eco-geographical diversification. Secondly, karyotype evolution is considered at the level of whole genus Leishmania. Despite the fast chromosome evolution rate, there is qualitative congruence with MLEE- and RAPD-based evolutionary hypotheses. Significant differences may be observed between major lineages, likely corresponding to major and less frequent rearrangements (fusion/fission, translocation). Thirdly, comparison is made with Trypanosoma cruzi. Again congruence is observed with other hypotheses and major lineages are delineated by significant chromosome rearrangements. The level of karyotype polymorphism within that "species" is similar to the one observed in "genus" Leishmania. The relativity of the species concept among these two groups of parasites is discussed.
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The invasion of the Bolivian Jungle has brought the new colonists some unfamiliar diseases, among which we study Leishmaniasis. A previous study described the situation in Yapacaní. Departamento of Santa Cruz, a primary rain forest lowland area. We now focus on the characteristics of Carrasco Tropical, close to a hilly territory of the andean mountains. We studied 11 localities ("colonies") grouped as unions with different lengths of residence in the area. We considered males and females over 15 years old as "at risk" and studied in them all forms of leishmanial infection, through clinical and laboratory (smears) means, including the Montenegro Skin Test (IDRM). Cutaneous ulcers and scars were seen in 2.9% (10 patients of 339 at risk, 6 from "27 de octubre", a younger settlement, 4 from the older Tamboradas): mucocutaneous lesions in 3 (1 from the younger settlement); and skin scars alone in 10.3% (35 from the younger area). The only 2 females with positive findings in the study were seen in this latter group. Transmission is apparently associated with the primary forest which exists at the foot of the Andes in the area, which is visited preferentially by young men.
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