[Popular nosology of infantile diseases in West Côte-d'Ivoire and consequences for malaria care management]. / Nosologie populaire des maladies infantiles dans l'Ouest de la Côte-d'Ivoire. Implications pour le paludisme.

In a rural area of intense and permanent malaria transmission in Southwest Côte-d'lvoire, traditional midwifes of the Yacouba ethnic group, with also an important function for the children health and care, were interviewed in 2002 about their knowledge of the infantile pathologies. Their nosology is greatly based on symptoms and etiologic explanations of the disorder of secretions. The accumulation of a viscous liquid in different parts of the body, especially in the chest, the throat or the head, explains numerous febrile or afebrile diseases, including malnutrition. Some pathologies, particularly convulsions, are described by analogy with animals behaviour Relations between these entities and the biomedical ones are difficult to establish. The traditional care and treatments result from these concepts. A vomiting child or a child suffering from diarrhoea is subject to devices to evacuate his excess of liquid. Yellow brews are used against jaundice. Furthermore, an important mistrust remains towards medical treatments particularly for all parenteral therapies. Health facilities are only used as a the last resort. Their bad reputation is confirmed by the high rate of mortality of patients coming often too late. To improve malaria care management, health-care workers have to take into consideration these concepts and also prove their abilities to ensure good medical practices.

[Influence of social perceptions and practices on the use of bednets in the malaria control programme in Ivory Coast (West Africa)]. / Representations sociales et pratiques liées à l'utilisation des moustiquaires dans la lutte contre le paludisme en Côte d'Ivoire (Afrique de l'Ouest).

The National Malaria Programme in Ivory Coast has encountered difficulty in winning public acceptance of insecticide-treated bednets. We speculate that resistance to the use of bednets could be rooted in social perceptions, beliefs and practices in the communities. The purpose of this study was to identify sociocultural and environmental factors that could be used to support promotion strategies and acceptance of impregnated bednets in Ivory Coast. Survey findings confirmed that bednets were not in widespread use among the population (25%). The most widely used methods were burning mosquito coils (50%) and indoor spraying (31%). Use of impregnated bednets was low (6%). Most survey respondents (73%) indicated initial appreciation for the effectiveness of bednets in protecting against mosquitoes as a nuisance. However only 9% of respondents thought that impregnated bednets provided protection against malaria although they did not necessarily use them. Design was a determinant factor for the use, and even acceptance, of bednets. The population want rectangular, permanently impregnated bednets large enough to accommodate at least 2 persons. Cost was a major obstacle to wider use by the population. According to our data the best price for the population would be between 2000 and 2500 FCFA as compared to the current price of 3500 FCFA in Ivory Coast.

Representations sociales et pratiques liees a l'utilisation des moustiquaires dans la lutte contre le paludisme en Cote d'Ivoire (Afrique de l'Ouest)

Devant les difficultes que rencontre le Programme National de Lutte contre le Paludisme (PNLP) de Cote d'Ivoire; dans sa volonte d'amener les populations a adopter les moustiquaires impregnees d'insecticide; nous nous sommes demandes si les representations sociales; les attitudes et les pratiques des communautes liees a l'utilisation des moustiquaires n'expliquent pas cette contre-performance. Notre etude a eu pour but de documenter les facteurs socioculturels et environnementaux pour renforcer les strategies de promotion et de vulgarisation de la moustiquaire impregnee en Cote d'Ivoire. Les resultats obtenus montrent qu'au niveau des enquetes quantitatives; la moustiquaire en general est faiblement utilisee par les populations (25). Les moyens les plus utilises sont les serpentins fumigenes (50) et les bombes aerosols (31). La moustiquaire impregnee est tres faiblement utilisee (6). La moustiquaire est; de maniere generale; appreciee dans un premier temps pour son efficacite dans la protection contre les nuisances dues aux moustiques par 73des enquetes. Seulement 9de ces enquetes pensent que la moustiquaire impregnee sert a se proteger contre le paludisme; mais ne l'utilisent pas necessairement. L'organisation des unites de couchage; voire leur inadaptation; sont determinantes dans l'utilisation de la moustiquaire. La moustiquaire souhaitee par les populations est celle d'au moins 2 places; de forme rectangulaire; de couleur blanche; faite en tulle a mailles fines; transparente; impregnee d'insecticide et a impregnation definitive. Toutefois; son cout semble etre le handicap majeur a son adoption par les populations; pour qui le cout ideal de la moustiquaire a l'achat; se situe entre 2000 et 2500 FCFA contre les 3 500 Frs actuellement pratiques en Cote d'Ivoire

[Environment and child health: from health transition to shared risk?]. / Environnement et santé de l'enfant: de la transition sanitaire au risque partagé?

Children under the age of 18 account for almost half of the world's population, with most living in developing countries. Young people are especially sensitive to acute and chronic environmental conditions and 43% of environmental diseases occur in the 12% of the world's population under age 5. The main environmental threats to the health of children in developing countries are inadequate access to clean water for drinking and hygiene, exposure to air pollution: primarily indoors and secondarily outdoors, risk of accidents and wounds, and poisoning due to toxic products. Recent data suggest that the number and diversity of environmental risk factors affecting child health is increasing as a result of increasing malnutrition, pollution, and violence and consequently that the level of health and quality of life of future generations will decrease. Due to the complexity of the interactions between environmental factors and socio-economic determinants, the epidemiological transition model is poorly suited to analyzing and predicting the concurring risks of infectious disease and chronic disease (diabetes, cancer...). This article presents a number of recommendations for training health professional, developing environmental reference centers, implementing risk assessment, coordinating decentralized activities and policy, and involving parents and children in the decisional process with emphasis on divulgating study findings and developing interfaces between the various stakeholders.

[Role of spleen cells in protection against malaria]. / Rôle des cellules de la rate dans la protection contre le paludisme.

IMMUNITY: The anti-infectious effect of the spleen in general and its anti-malarial effect has been known for a long time. This effect is associated, among other functions, with the spleen's capacity to filter and destroy erythrocytes parasitized by plasmodies or altered by various physical (heat...) or chemical factors. In addition, splenic immunity, which associates specific and nonspecific humoral and cellular effector mechanisms is no longer a question of debate. OPEN QUESTIONS: There remains nevertheless one aspect of the question to be elucidated: the probable interaction between this immunity and other (genetic...) factors of the malarious host. CELL FUNCTIONS: This article summarizes current knowledge in connection with: a) participation of various cell populations in the mechanisms of splenic filtration and phagocytosis and b) genesis of a repertory of B and T lymphocytes, plasmodio-specific memory cells. During an infection, lymphocytes, monocytes and macrophages are recruited from peripheral blood and, in cooperation with cells known as "barrier" cells, increase the capacity of splenic filtration/purge and phagocytosis. In addition, the appearance of B and T lymphocytes with specific memory of P. falciparum result from hypermutation in VH genes (for B lymphocytes) or from clonal selection (for T lymphocytes). The folicular dendritic cells accomplish a reserve function. By constantly releasing the antigen, these cells would contribute to maintaining immune memory or to stimulating naive cells. Further studies are necessary to better understand the role of the splenic microcirculation and to identify parasite components which stimulate the protective response of the spleen against plasmodies.

Partial protection of mice against Trypanosoma cruzi after immunizing with the TcY 72 antigenic preparation.

A 72 kDa Trypanosoma cruzi glycoprotein recognized by the 164C11 monoclonal antibody (IgM isotype) was purified by preparative electrophoresis. The antigenic preparation obtained, named TcY 72, was used to immunize C57Bl/10 mice. The following results were observed after immunization: (1) induction of higher titres of IgG than IgM antibodies, as evaluated by indirect immunofluorescence; (2) significant DTH after injection of epimastigotes in mice footpads; (3) peak parasitemia in immunized mice was significantly reduced and animals were negative by 13 days post-infection, although the mice still succumb to infection; (4) the phenotypic analysis of spleen cell populations showed a decrease in the CD4/CD8 ratio in immunized mice. Taken as a whole, these findings indicate that TcY 72 is immunogenic and potentially important for protective immunity.

Antigenic components of partially purified antigens of Leishmania donovani infantum recognized by sera from dogs with asymptomatic or active visceral leishmaniasis.

The antigenic components of a semipurified fraction of Leishmania donovani infantum were determined by sodium dodecyl sulfate-polyacrylamide gel electrophoresis and Western blot analysis using 14 serum samples from dogs with symptomatic visceral leishmaniasis and 11 serum samples from apparently healthy dogs collected in an area endemic for canine visceral leishmaniasis (CVL). It was found that these antigens were composed of many polypeptides, among which seven components recognized by symptomatic CVL sera, had molecular weights of approximately 18, 28, 30, 33, 63, 70, and 72 kilodaltons (kD); two components of 63 and 70 kD were recognized by three of 11 healthly dog sera. These findings suggest that specific antigens induce humoral immune response in dogs with asymptomatic or active visceral leishmaniasis. Infected dogs are not readily identifiable by their symptoms. The potential interest of the immunoblot test for CVL diagnostic purposes is discussed.

Antigenic components of excretory-secretory products of adult Fasciola hepatica recognized in human infections.

The antigenic components of excretory-secretory products (ESP) of adult worms of Fasciola hepatica were revealed by sodium dodecyl sulfate-polyacrylamide gel electrophoresis and Western blot analysis using sera from 20 patients infected with F. hepatica. Sera from 184 other parasitic infections and 20 healthy volunteers were also analyzed. It was found that the ESP were composed of more than 11 polypeptides; five components detected in fascioliasis sera had molecular weights of 12.4, 16.4, 19.4, 25, and 27 kilodaltons (kD). Only the 25- and 27-kD components were recognized by all 20 fascioliasis sera. Using the ESP as antigen, it was possible to perform an enzyme-linked immunosorbent assay with a sensitivity of 95% and a specificity of 97%. Sera from other parasitic infections had antibodies to antigenic components with apparent molecular weights of 37, 38.4, 52, 63, 73, 87, 109, and 116 kD that were also found in sera from fascioliasis patients. These findings suggested that the 25- and 27-kD antigenic components may be sensitive and specific for the diagnosis of human fascioliasis.

Canine visceral leishmaniasis: successful chemotherapy induces macrophage antileishmanial activity via the L-arginine nitric oxide pathway.

Following successful chemotherapy in canine visceral leishmaniasis, monocyte-derived macrophages can induce antileishmanial activity via a gamma interferon-dependent mechanism in the presence of autologous lymphocytes. The killing of leishmania correlated with the induction of the NO synthase pathway, because it correlated with the generation of nitrogen derivative production and was abrogated in the presence of NG-monomethyl-L-arginine, a competitive inhibitor of the NO synthase pathway. The level of L-citrulline in serum, which was produced after activation of the NO synthase pathway, was markedly enhanced in dogs receiving successful chemotherapy. Taken together, these data indicate that following successful chemotherapy of visceral leishmaniasis, leishmania parasites are killed by macrophages activated by gamma interferon-producing lymphocytes via an NO-dependent mechanism.

A monoclonal antibody against blood forms of Trypanosoma cruzi lyses the parasite in vitro and inhibits host cell invasion.

Monoclonal antibodies (mAbs) of the IgM isotypes were produced from mice immunized with blood forms of Trypansoma cruzi Y strain. Characterization of the epitope recognized by one of the mAbs, 164C11, as well as the effects of this mAb on complement-mediated lysis and host cell invasion are reported. Immunocytochemical analysis showed that the mAb was reactive with various strains of T. cruzi (Y, WSL, and Colombiana) as well as other trypanosomatids. The mAb 164C11 demonstrated a high complement-mediated lytic activity against bloodstream trypomastigotes, being more effective than chronic mouse serum. A protein with an apparent molecular weight of 72 kDa was detected by this mAb on all developmental stages of T. cruzi. Studies using periodate and endoglycosidase treatments suggested that the epitope is not a carbohydrate and seems to be located on the parasite membrane. In addition, preliminary results are presented, suggesting that the 72-kDa protein is involved in adhesion/or internalization of bloodstream trypomastigotes.

In vitro study of the anti-leishmanial activity of biodegradable nanoparticles.

Leishmania are obligate intracellular parasites, responsible for leishmaniasis. Leishmaniasis are transmitted via insect vector to vertebrate hosts including humans. The infection was reproduced in vitro with promastigotes which can infect murine resident peritoneal cells. Amphotericin B was incorporated into poly(D, L-lactide-co-glycolide) nanoparticles, biodegradable drug carriers, to allow specific targeting inside the cell. The interaction of the drug with infected cells was determined by exposing macrophage cultures to drug carriers. The toxic effects of polymeric drug carriers were defined prior to exposing cells to drug-loaded nanoparticles. For contact times up to 4h, cells tolerated polymer concentrations of 0.01%. The viability of parasites after treatment was determined. Infected macrophages were incubated at 26 degrees C (which allows the transformation of amastigote to promastigote) along with loaded and unloaded nanoparticles, as well as the free drug alone, and a count of the parasites in the medium was recorded. Anti-leishmanial activity was observed with drug-free nanoparticles. This activity may arise through the release of hydrogen peroxide following the activation of macrophages. The incorporation of amphotericin B did not enhance this effect. Interestingly, trehalose, a cryoprotector of the freeze-dried nanoparticles, altered parasite growth and activated macrophages.

Exploitation of parasite-derived antigen in therapeutic success against canine visceral leishmaniosis. Veterinary Group of Lupino.

In an attempt to obtain therapeutic success against canine visceral leishmaniosis, the potential of LiF2 antigen (Leishmania infantum-derived Fraction 2, 94-67 kDa), given alone or in combination with the chemotherapeutic agent N-methylglucamine antimonate, was compared with conventional chemotherapy with that drug. Absence of any parasite in direct microscopic examination of bone-marrow aspirates in treated dogs was considered a parasitological cure, i.e. therapeutic success. Results showed that the disappearance of clinical symptoms did not always indicate parasitological healing in dogs. The parasitological healing rates with chemotherapy and immunotherapy alone were 37.5% and 25% respectively, in contrast to the 100% cure rate observed with chemotherapy combined with immunotherapy. The development of a protective response in dogs, as measured by the in vitro leishmanicidal activity of monocyte-derived macrophages in the presence of autologous lymphocytes, was found to correlate well with the success of therapy. The overall findings of this study give an important insight into the immunotherapeutic strategy by which therapeutic success can be achieved in canine visceral leishmaniosis.

Exploitation of parasite derived antigen in therapeutic success of human cutaneous leishmaniasis in Brazil.

In a complete study in 25 patients with American cutaneous leishmaniasis, caused by Leishmania braziliensis complex, immunotherapeutic efficacy of parasite derived antigen (94-67 KD) has been compared to antimonial therapy. Additionally, to delineate the mechanism of therapeutic success, microscopical features of immune response in active lesions and healed or non-healed lesions following therapy were analyzed. The results showed that cure rates in immunotherapy and chemotherapy were equal (> 83%). The immunohistochemical changes in two therapeutic groups were also largely similar. The analysis of humoral and cellular immune response suggest that appropriate stimulation of T helper cells in the lesion site, in association with one or more cytokines, play a key role in the healing process.

[Fixation and immunological reactivity of parasitic protozoa in sol-gel matrices]. / Fixation et réactivité immunologique de protistes parasites au sein de matrices sol-gel.

Parasitic protozoa have been entrapped within sol-gel silica matrices. Stationary phase promastigote cells of Leishmania donovani infantum are mixed with a silicon alkoxide solution. They remain trapped within the growing oxide network formed upon hydrolysis and condensation of the alkoxide. Electron microscopy shows the preservation of the cellular organization and the integrity of the plasma membrane of entrapped parasites. Specific antigen-antibody reactions have been performed within the sol-gel matrix via enzyme-linked immunosorbent assays (ELISA) currently used for the diagnostic of visceral leishmaniasis. A clear-cut difference in optical density was measured between positive and negative sera from dogs. Entrapped protozoa antigens are still able to react with specific antibodies through the open porosity of the silica network. These results suggest that sol-gel matrices could be used for the development of immunodiagnostic assays requiring whole cell parasites as antigens.

CD23 and IgE expression during the human immune response to cutaneous leishmaniasis: possible role in monocyte activation.

Leishmania brasiliensis causes cutaneous leishmaniasis (CL) in humans. During this infection, a variety of inflammatory mediators are produced by T cells and monocytes/macrophages. In the present study, we analysed serum IgE levels and their correlation with in situ expression of the low affinity receptor for IgE (Fc epsilon RII/CD23) in patients infected with L. brasiliensis before and following therapy. These analyses were compared to in situ expression of tumour necrosis factor-alpha (TNF alpha), interleukin 3 (IL3), interferon-gamma (IFN gamma) and IL4. Disease-free individuals from the same endemic area sensitized with L. brasiliensis antigens were also included in this work. Our data indicate that during infection, serum levels of IgE and TNF alpha increased and correlated with elevated in situ expression of CD23, IL4 and TNF alpha mRNA. This expression disappeared following successful treatment, but persisted in patients resistant to anti-leishmania therapy. Patients resistant to therapy differed from other cases by a dramatic decrease in their in vivo expression of IFN gamma protein. Analysis of CD23 function in purified human monocytes indicated that this antigen mediates IgE/anti-IgE-dependent TNF alpha production. These data suggest a possible in vivo role of CD23 in acute immune responses in human CL.

Cytokines and T-cell response in malaria.

The intracellular protozoan Plasmodium sp induces a complex immune response which sometimes implies serious pathological effects for the host. According to in vitro studies and epidemiological surveys, several effector mechanisms are displayed against plasmodial blood stages and a large interaction between humoral and cell-mediated immunity is presumed to occur among protected individuals. The key role of T cells in the antiplasmodial immune response is now well established, but all the regulatory heterogenous mechanisms are not yet fully known. An increasing body of data shows a dual role during malaria attack for some cytokines released by monocytes and macrophages (TNF, IL-1, IL-6) or by T cells (IFN-gamma, lymphotoxin (LT), IL-4). The importance of some plasmodial proteins in the cytokine-induced pathology and the stimulation of a preferential TH1 or TH2 mediated immune response to achieve protective immunity against Plasmodium sp are discussed.

[Cell-mediated immunity and protection against blood stages of Plasmodium falciparum]. / Immunité à médiation cellulaire et protection contre les stades sanguins de Plasmodium falciparum.

In recent years, cell-mediated immunity against malaria has been the subject of intensive investigation either in humans from malaria endemic areas, or experimental models. Cellular immune mechanisms have been regarded as secondary to humoral immunity but, there is increasing evidence that shows its critical role in protection against blood stage plasmodium parasites. In the context of a large humoral-cellular interaction, T helper lymphocytes and monocytes/macrophages may play a key role in the elimination of plasmodial blood stages, particularly P. falciparum. IL-2, IL-4, IL-5, IFN-gamma cytokines secreted principally by CD4+ T lymphocytes and oxygen and nitrogen radicals produced by activated macrophages, are involved in the control of plasmodial infection. The spleen also plays a very important function in the anti-malarial protection by its increased capacity for filtration/destruction of parasitized red blood cells and by induction of B and T memory lymphocytes. Successful vaccination against malaria needs a choice of plasmodial antigens or B and T immunodominants epitopes able to stimulate plasmodium-specific lymphocytes and functional modification in the spleen.

[Soluble antigens of intra-erythrocyte stages of Plasmodium falciparum: diagnostic and vaccinal value]. / Les antigènes solubles des stades intra-érythrocytaires de Plasmodium falciparum: intérêt diagnostique et vaccinal.

A wide variety of soluble antigens also called exoantigens or excretory-secretory antigens are released by asexual blood stages of P. falciparum at the time of schizont burst and subsequent merozoite invasion. At present, most of these soluble malarial proteins have been purified and have had their genes cloned; the primary sequence displays considerable polymorphism upon tandem repeats. However, its function and the relevance of the polymorphism for the induction of host immune response, predominantly IgM and T-cell-independent type, has yet to be determined. The potential of P. falciparum exoantigens as immunodiagnostic tools has been the focus of numerous studies. Enzyme immunoassays or radioimmunoassays for the detection of malarial antigens in blood of suspected peoples have been reported. Their development requires increasing both specificity and sensitivity. Recently, some P. falciparum exoantigens have been proposed as candidates for inclusion in an anti-disease vaccine which induces a clinical but not parasiticidal immunity against malaria. There is much interest in this concept and their efficiency remains to be confirmed, as new strategies are needed in the prevention of malaria.

Serodiagnosis and screening of canine visceral leishmaniasis in an endemic area of Corsica: applicability of a direct agglutination test and immunoblot analysis.

For effective control of visceral leishmaniasis caused by Leishmania infantum in the Mediterranean area, the detection of infected dogs is of utmost importance. To assess the suitability of a direct agglutination test (DAT) and immunoblot analysis in serodiagnosis and screening of infected dogs under field conditions, a study was performed on 113 dogs in an endemic area of Corsica. Twenty one of 22 parasitologically confirmed cases were correctly diagnosed by both tests, and 100% specificity was found when 11 dogs with other diseases were examined. Interestingly, eight of 80 apparently healthy dogs from the same area were found to be parasite-positive by the DAT test as well as by the immunoblot. Although both tests were equally sensitive and specific, based on both the feasibility of its application in field conditions and ease of performance, we consider the DAT to be more suitable for serodiagnosis and large-scale screening of infected dogs.