Search PubMed⌕ Search

Biomedical subjects

A Capron

Publications and source records attributed to A Capron.

At least 379 records · Page 21Linked to original sources

Requirements of defined cultivation conditions for standard growth of Leishmania promastigotes in vitro.

The growth characteristics of L. chagasi (MHOM/BR/79/LI01) and L. braziliensis (MHOM/BR/72/1670), the causative agents of visceral and muco-cutaneous leishmaniases, respectively, were compared. Inoculum size clearly influences the growth course of both Leishmania species, whatever the culture medium used (serum-supplemented media: GLSH or RPMI, and a chemically defined medium: LITR9). Cultures initiated with low concentrations failed to promote cell growth, while typical growth curves were obtained when higher promastigote inocula were used. For all the species tested, the higher the initial density of flagellates in the medium, the shorter were the periods covered by the latent and particularly by the logarithmic growth phases. In contrast, using constant inocula, variations in the volume of the incubation medium did not change the time-course of the different culture phases of either Leishmania species, provided that the ratio of incubation medium to total flask volume was comparable. Only cell division time significantly increased with the culture volume. We also determined whether or not the growth characteristics of the promastigotes of L. chagasi or L. braziliensis could be generalized to other members of the genus. Our results show that, whatever the culture medium used, L. infantum behaves in the same way as does L. chagasi, whereas L. panamensis, L. guyanensis, L. mexicana and L. amazonensis display growth patterns similar to that of L. braziliensis.

Animals↗

Recognition of Toxoplasma gondii excreted and secreted antigens by human sera from acquired and congenital toxoplasmosis: identification of markers of acute and chronic infection.

While the serological response to somatic antigens of Toxoplasma gondii is currently analysed, little information is available on the antibody response to the antigens excreted and secreted by tachyzoïtes (ESA). This serological study is focused on the immune response towards these antigens which were released by the parasites in cell-free culture medium. Human sera corresponding to 'acute', 'subacute' and 'chronic' acquired infection and sera from infected newborns and from their mothers were analysed by radio-immunoprecipitation with 35S methionine-labelled ESA and with radio-iodinated membrane antigens followed by polyacrylamide gel electrophoresis. In chronic toxoplasmosis, IgG antibodies recognized among ESA major 108, 97, 86, 60, 57, 42, 39 and 28.5 kD antigens; the 108-97 kD doublets and the 28.5 kD antigen seemed characteristic of the chronic phase of toxoplasmosis. In acute infection, IgM antibodies to the 97 kD antigen, the first to appear, seem to contitute good markers of early acute infection. The comparative study of antibody response to membrane antigens showed that, in chronic toxoplasmosis, human sera recognized four antigens of 43, 35, 30 and 22 kD and that, in acute toxoplasmosis, they first recognized the 43 and 30 kD antigens. The serological evolution in congenital toxoplasmosis was the same as in acquired infection. In some cases, the serological profile of the newborn was different from that of his mother, with an additional antibody response to a 170 kD antigen. This study demonstrates in human toxoplasmosis an early, intense and characteristic antibody response against ESA, suggesting that the use of these antigens could lead in the future to improved diagnostic tests.

Acute Disease↗

Eosinophilic gastroenteritis: ultrastructural evidence for a selective release of eosinophil major basic protein.

Ultrastructural study of mucosal eosinophils in a case of eosinophilic gastroenteritis involving stomach, duodenum and ileum showed an altered structure in ulcerated duodenal areas. The electron core density of eosinophil granules was inverted or disappeared and tubulovesicular structures occurred. Using immunogold staining with specific antibodies, major basic protein was detected diffusely in the matrix of eosinophil granules and out of the granules in tight association with extragranular membrane formations. In contrast, eosinophil cationic protein and eosinophil peroxidase were normally distributed in the granule matrix. When compared with the eosinophils in macroscopically normal duodenal mucosa in the same patient, these changes support a role for major basic protein in tissue damage in eosinophilic gastroenteritis. The diffusion of one granule protein from the granules to the exterior of the cells favours the view of a selective release of eosinophil mediators.

Adult↗

Lymphocyte-mediated regulation of platelet activation during desensitization in patients with hymenoptera venom hypersensitivity.

T cells from peripheral blood of hymenoptera sensitive patients were studied before and after venom desensitization. Before treatment, T cells showed a variable but higher proliferative response to allergen than T cells of treated patients or controls. While before desensitization, T cell products, specifically released after in vitro allergen stimulation, were able to amplify the IgE-dependent platelet activity, we showed that after treatment of the same patients, T cell products strongly reduced platelet activation. Considering the modifications in platelet activation previously observed in patients treated by specific immunotherapy, the present results suggest that, through a modification of T cell reactivity to allergen, T cell functions are modulated by desensitization, and emphasize the involvement of T cell products in the desensitization mechanisms.

Adolescent↗

Lymphatic filariasis: detection of circulating and urinary antigen and differences in antibody isotypes complexed with circulating antigen between symptomatic and asymptomatic subjects.

A two-site immunoradiometric assay using a monoclonal antibody (MoAb) against Brugia malayi microfilariae allowed the detection of parasite molecules both in the serum and the urine of patients from Sri Lanka infected with Wuchereria bancrofti. Whereas 50% of patients had no antigen in their serum, all of them excreted detectable amounts of antigen in their urine, the levels being higher in symptomatic than in asymptomatic patients. The poor detection in serum appeared to be related to the presence of circulating immune complexes. It was shown that the isotype of the antibodies complexed with the circulating antigen was IgM in the asymptomatic group, while it was mainly IgG in the symptomatic patients (swelling and lymphoedema or elephantiasis). These results suggest the existence of regulatory immune mechanisms affecting the clinical expression of lymphatic filariasis.

Animals↗

Decrease of IgE-dependent platelet activation in Hymenoptera hypersensitivity after specific rush desensitization.

A receptor for the Fc fragment of IgE on human platelets has been recently described, which mediated an IgE-dependent activation in the presence of specific allergen. We investigated the allergen-induced activation of platelets from patients with Hymenoptera hypersensitivity before and after specific rush desensitization. Nineteen patients with a history of anaphylactic reactions were included (15 sensitive to yellow-jacket and four to honey-bee venom), fourteen/nineteen having experienced severe life-threatening systemic reactions and 5/19 large local reactions. All showed positive skin tests and high values of specific IgE. By comparison to the baseline results obtained before desensitization, a significant decrease of platelet activation (76.8% inhibition) after rush desensitization was observed. In the case of two polysensitized patients, after Hymenoptera venom desensitization alone, platelets not only lost their reactivity to venom but also towards the other allergen. This modulation of the IgE-dependent platelet reactivity during desensitization offers therefore a new approach for the study of allergen-induced desensitization.

Adolescent↗

Immunity to schistosomes: progress toward vaccine.

Among the major parasitic infections, schistosomiasis may be the most promising candidate for human vaccination. Information about mechanisms of immunity, gained mainly from experimental models but likely to be relevant to human infection, indicates a dynamic balance between protective and regulatory (blocking) mechanisms. Besides cell-mediated responses leading to macrophage activation, antibody-dependent cell-mediated cytotoxicity systems involving precise antibody isotypes and nonlymphoid cells (mononuclear phagocytes, eosinophils, and platelets) appear to be essential effectors of immune attack. The slow development of immunity in humans seems related to the production of antibodies that cross-react with schistosomulum surface antigen and block the binding of antibodies of the effector isotype. Schistosomes that survive in the bloodstream and produce chronic infections may evade the immune system as a result of intrinsic changes in membrane susceptibility and of transient expression of target antigens; at other stages of the parasite life cycle, cross-reactive molecules may be secreted that play an essential role in the induction of immunity. Several schistosome proteins have been characterized as candidates for vaccination. Among these, an antigen of 28 kilodaltons has been cloned and shown to be immunogenic in humans and protective in mice, rats, and baboons.

Animals↗

Functional role of the alpha-chain of complement receptor type 3 in human eosinophil-dependent antibody-mediated cytotoxicity against schistosomes.

The participation of complement receptor type 3 (CR3) in antibody-dependent effector function of human eosinophils against parasites was studied by using monoclonal antibodies directed against various surface molecules. Both adherence and cytotoxicity of hypodense eosinophils to IgE-coated schistosomula of Schistosoma mansoni were strongly inhibited by anti-CR3 antibodies (OKM1 or Mo1). The specificity of the inhibitory effect for the alpha-chain of CR3 was shown by the lack of inhibition of anti-beta-chain or anti-LFA1 alpha-chain monoclonal antibodies, although these antigens were expressed on human eosinophils. These results associated to previous works on IgE receptors demonstrate that both receptor for Fc fragments of IgE and CR3 are essential in IgE-dependent cytotoxicity of human eosinophils. Flow microfluorometry analysis revealed that hypodense eosinophils were more intensively stained by OKM1 antibodies than the normodense populations. In the case of IgG-mediated cytotoxicity by normodense eosinophils, only the enhancement of cytotoxicity due to monokine activation was inhibited by anti-CR3 alpha-chain antibodies. These findings suggest an increased expression of CR3 on eosinophils after activation either in vivo or in vitro. The participation of CR3 in IgE-mediated cytotoxicity against schistosomes was also required in the case of blood monocytes but not for platelet-mediated killing, which does not require prior adherence. The biologic role of CR3 is therefore extended to effector mechanisms involving eosinophils and two different isotypes of antibodies and possibly implied in immunity against schistosomes.

Antibodies, Monoclonal↗

Compound 48/80 is a potent inhibitor of phospholipase C and a dual modulator of phospholipase A2 from human platelet.

Compound 48/80 inhibited phosphatidylinositol-specific phospholipase C activity from human platelets. Whereas 1 microgram/ml of compound 48/80 slightly stimulated Ca2+-dependent phospholipase A2, higher concentrations led to dose-dependent inhibition of this platelet enzyme. This biphasic effect was confirmed with phospholipases A2 purified from rat liver and human synovial fluid. The aggregation of human platelets induced by ADP and PAF-acether was inhibited by compound 48/80, whereas the aggregation induced by ionophore A23187 was not modified by this compound. These results demonstrate that the inhibition of platelet aggregation by compound 48/80 is not due solely to effects on calmodulin as previously reported, but that inhibition of phospholipases and probably arachidonate mobilization may also be involved.

Animals↗

Functional properties of a rat monoclonal IgE antibody specific for Schistosoma mansoni.

A rat monoclonal antibody of IgE isotype (B48-14) raised against Schistosoma mansoni has been generated by the fusion of mesenteric lymph node cells from LOU/M rats immunized with a preparation of adult schistosome worms and IR973F nonsecreting rat myeloma cells. Investigation of the in vitro effector functions of this IgE antibody showed a high level of cytotoxicity against S. mansoni schistosomula in the presence of eosinophils, macrophages, and platelets. A significant level of protection (40 to 60%) against a challenge infection with S. mansoni cercariae was achieved by passive transfer experiment of B48-14 IgE to naive recipient rats. By immunoprecipitation, B48-14 IgE antibodies were shown to react with an antigen of 26 kDa present in excretion-secretion products of schistosomula, previously described as a potential immunogen eliciting a protective IgE response against schistosomiasis.

Animals↗

Induction of platelet cytotoxic functions by lymphokines: role of interferon-gamma.

Antigen- or mitogen-stimulated CD4+/CD- lymphocytes produced factors able to induce normal human platelets into cytotoxic effectors toward the young larvae of Schistosoma mansoni. The neutralization by monoclonal anti-IFN-gamma antibody of the induction of the platelet killer effect, the presence of IFN-gamma in the CD4+/CD8- lymphocyte supernatant, and, finally, the direct inducer effect of recombinant IFN-gamma clearly demonstrated that IFN-gamma was one of the factors responsible for the induction of platelet cytotoxic functions.

Adult↗

[Visceral lesions in hypereosinophilia].

The hypereosinophilic syndrome is an ill-defined nosological entity with predominant risks of cardiac and/or neurological lesions. In the light of new data on the effector cytotoxic effects of eosinophils, we have tried to establish new criteria of severity by purifying the circulating eosinophils of 14 patients with hypereosinophilic syndrome and testing their toxicity. This study has revealed the existence of low density ("hypodense") eosinophils with potential cytotoxicity in vitro. The worst clinical forms of the syndrome were observed in the group of patients with positive eosinophil toxicity tests. The significance of these cellular changes (hypodensity, eosinotoxicity) and their relationship with the clinical manifestations are discussed.

Adult↗

A purified 28,000 dalton protein from Schistosoma mansoni adult worms protects rats and mice against experimental schistosomiasis.

We have purified a 28,000 dalton (P28) protein from Schistosoma mansoni adult worms and used it to immunize Fischer rats. Immunofluorescence assays demonstrated that the P28 antigen was mainly located in the parenchyma of the schistosomulum and of the adult worm, including the dorsal spines of the parasite. Western blot analysis revealed that this antigen was present in three species of schistosomes: S. mansoni, S. japonicum, and S. bovis. The antibody response raised against this protein was able to kill S. mansoni schistosomula in in vitro cytotoxicity assays in the presence of rat eosinophils. The inhibition of this cytotoxic activity by an aggregated myeloma IgG2a indicated that one of the major isotypes involved in this in vitro model is IgG2a. The passive transfer of P28 antisera induced a significant level of protection against experimental infection. Moreover, we have immunized Fischer rats and BALB/c mice with the purified 28,000 dalton protein and observed a marked decrease (up to 70%) in the parasite burden in both experimental infection models.

Animals↗