Biomedical subjects

A Capron

Publications and source records attributed to A Capron.

At least 73 records · Page 4Linked to original sources

PubMed1998

Toxoplasma gondii: kinetics of the dissemination in the host tissues during the acute phase of infection of mice and rats.

Mice and rats differ in their susceptibility to Toxoplasma gondii infection. Here we have compared the parasitological parameters of acute infection in both mice and rats infected either orally with cysts of Prugniaud strain or intraperitoneally (ip) with tachyzoites of the RH strain. The animals were killed at regular interval during the acute phase, and the parasitic burden in mesenteric lymph nodes, spleen, liver, diaphragm, heart, lungs, brain, and blood was assessed by a tissue culture method in MRC5 fibroblast cells. Mice infected with the RH strain showed a drastic increase of the parasitic load in all organs (up to 10 (8) parasites / g of organs), and then died. When mice were infected with 50 cysts of Prugniaud strain, parasites were first observed in mesenteric lymph nodes, spleen, and lungs and then in other organs. In rats, infection with 1200 cysts of the same strain led to a similar pattern; however, the chronology of the infected organs changed when they were infected with RH strain tachyzoites. These results suggest that the parasites, present first in the peritoneal cavity in the case of RH ip infection, infect all the adjacent organs and then the blood which disseminates the tachyzoites all over the organism. In contrast, after an oral infection, the parasite crosses the intestinal barrier to reach the mesenteric lymph nodes and then the spleen and are finally distributed by the blood throughout the organism. We also showed that following infection with a lethal or a sublethal doses of the Prugniaud strain, the parasitic burden in the studied organs was similar and therefore does not directly correlate with the death of the mice.

Acute Disease↗

PubMed1998

Molecular cloning and expression of an anti-idiotype antibody mimicking a protective oligosaccharide of the parasite Schistosoma mansoni.

Genes encoding the heavy and light chains of an anti-idiotype antibody (AB2) mimicking a protective oligosaccharide of Schistosoma mansoni were cloned and expressed as a single-chain Fv fragment. The expression in a functional state was tested using the AB1. A specific binding between sFv and AB1 was observed. Immunization with the recombinant AB2 indicates its capacity to elicit anti-S. mansoni antibodies.

Amino Acid Sequence↗

PubMed1998

Schistosomiasis: forty years' war on the worm.

Schistosome vaccine candidates are now entering phase I clinical trials. In an attempt to illustrate the progress made towards control, and the definition of potential vaccines against schistosomiasis, André Capron reviews some of the major findings and observations that have led to a significant evolution of our concepts, together with the production of novel and promising tools. It is obvious that our most important advances have proceeded from the successful integration of convergent knowledge in biology, epidemiology and biotechnology. At this stage, nobody can predict what will be the outcome of the current trials, but an important step has certainly been made towards immunointervention in human schistosomiasis.

Journal Article↗

PubMed1998

Alterations in cytochrome-c oxidase expression between praziquantel-resistant and susceptible strains of Schistosoma mansoni.

The genetic differences between praziquantel-resistant (R) and susceptible (S) strains of Schistosoma mansoni (Fallon & Doenhoff, 1994) were explored using RAPD and by cloning differentially expressed mRNAs by subtractive PCR. No differences between the 2 strains were detectable by RAPD using 41 different primers indicating that no major genomic rearrangements were present. Subtractive PCR generated a number of fragments, 1 of which was shown to correspond to an over-expressed mRNA in the R strain and to encode a fragment of the subunit 1 of cytochrome-c oxidase (SCOX1). In the absence of a complete sequence for this gene, we used EST sequences to compile a consensus sequence for the 904 bp at the 3' end that enabled us to choose primers for semi-quantitative RT-PCR. This technique showed that SCOX1 was indeed over-expressed about 5 to 10-fold in the R strain whereas the genes encoding the 28 kDa glutathione S-transferase, glutathione peroxidase, NADH dehydrogenase subunit 5 and the ATP-binding cassette family protein SMDR2 were not. In contrast, cytochrome-c oxidase enzyme activity was 4-fold lower in the R strain than in the S strain.

Amino Acid Sequence↗

PubMed1998

Homologous and heterologous protection after single intranasal administration of live attenuated recombinant Bordetella pertussis.

While single-dose mucosal immunization is best achieved by the use of attenuated live microorganisms, attenuation generally results in decreased immunogenicity. We attenuated Bordetella pertussis by the deletion of the pertussis toxin gene. A single intranasal administration of this strain protected against subsequent challenge as well as did the parent strain and better than immunization with commercial vaccine. Unexpectedly, this attenuation resulted in increased immunogenicity against the protective antigen filamentous hemagglutinin (FHA). In addition, immunogenicity was also enhanced against the Schistosoma mansoni Sm28GST genetically fused to FHA, resulting in protection against the parasite, as characterized by a reduction in worm burden and egg charge, after a single intranasal administration. Thus, attenuated recombinant B. pertussis strains are promising vectors for the simultaneous protection against pertussis and heterologous diseases by a single intranasal administration.

Adhesins, Bacterial↗

PubMed1998

Recombinant human IL-16 inhibits HIV-1 replication and protects against activation-induced cell death (AICD).

The chemoattractant cytokine IL- 16 has been reported to suppress lymphocyte activation and to inhibit HIV-1 replication in acutely infected T cells. We have cloned and expressed human IL-16 in Escherichia coli and investigated whether the recombinant protein could regulate the level of lymphocyte apoptosis from HIV-1-infected subjects. After purification and refolding, only 2-10% of the recombinant cytokine was present in a biologically active homotetrameric form. This could explain the need for high concentrations of the bacterially derived IL- 16 to induce significant inhibition of HIV-1 replication. Addition of IL-16 to unstimulated peripheral blood mononuclear cell (PBMC) cultures from HIV-1-infected subjects did not modify the observed level of spontaneous lymphocyte apoptosis. In contrast, IL-16 added to PBMC cultures stimulated with anti-CD3, anti-CD95 or dexamethasone reduced significantly the percentage of lymphocytes undergoing AICD. This effect was found to correlate with the ability of the cytokine to decrease CD95 expression on activated CD4+ T cells. Comparative studies on PBMC from healthy individuals indicated that the regulation of apoptosis levels by IL-16 is a complex phenomenon and could depend on the nature of the activator used and/or the immune status of lymphocytes tested. The outcome of CD4 cross-linking on T cells by various ligands is discussed in the context of the observed beneficial activities of IL- 16 and its potential role in the treatment of HIV disease.

Apoptosis↗

PubMed1998

In vivo expression of cytokine mRNA in rats infected with Schistosoma mansoni.

As an animal model, rat schistosomiasis mansoni has provided considerable knowledge of immune mechanisms involved in the expulsion of worms and in a subsequent development of immunity to reinfection. Although it is clear that ADCC mechanisms participate in immunity to reinfection; the nature of the cytokines involved in immunity is unknown. To analyse the pattern of cytokines involved, the mRNA levels of different cytokines were assessed by RT-PCR as they occur within tissues during the course of infection. In spleens from infected rats, a significant elevation in IL-2 and IL-5 mRNA was observed during the early phase of infection (day 7). Analysis of pulmonary cytokine responses showed a dramatic increase in IL-4 and IL-5 on day 7. This was accompanied with a low but significant increase in IL-2 (day 11) and IL-12 (day 7) in the absence of augmented IFN-gamma expression. The cytokine expression patterns of draining lymph nodes (LN) from infected rats showed a significant increase of IL-2, IL-4 and IL-5 on day 21. Analysis of IL-10 expression showed exclusively a significant increase in LN on day 11, IFN-gamma mRNA was not detected in any tissue sample. Thus, rats develop a predominately Th2-type cytokine response during a primary infection which may be involved at least in part, in the expression of immunity against Schistosoma mansoni infection.

Animals↗

PubMed1998

Murine Schistosoma bovis infection: analysis of parasitic and immune parameters.

Humoral and cellular responses to Schistosoma bovis antigens have been evaluated over a period of 11 weeks in mice exposed to S. bovis cercariae and data analysed in the context of the parasitic parameters (worm and egg loads) recorded at days 30, 60 and 80 of the ongoing infection. Results revealed a decrease of worm burden, particularly marked for female worms, between day 60 and day 80 of infection suggesting a higher susceptibility of female schistosomes to attrition mechanisms. The B-cell response, studied by measuring the production of different isotypes, was directed against different stage specific antigens, with a predominance of IgG1 antibodies associated with a significant increase of IgA and IgE antibodies after egg deposition. The T-cell response, assessed after in vitro stimulation of splenocytes, showed a predominant production of Th-2 cytokines (IL-4, IL-5 and IL-10) occurring after egg laying. Interestingly in contrast to S. mansoni infection the Th-2 polarization did not seem to be exclusively triggered by egg-associated antigens since significant amounts of IL-10 were produced after stimulation with adult worm antigen preparation (SWAP) before the beginning of egg deposition.

Animals↗

PubMed1998

Adjuvant is the major parameter influencing the isotype profiles generated during immunization with a protein antigen, the Schistosoma mansoni Sm28-GST.

We have previously shown that immunization of mice with the vaccine candidate, the 28-kDa glutathione-S-transferase of Schistosoma mansoni (Sm28-GST), in alum or complete Freund's adjuvant, or with recombinant Salmonella typhimurium expressing Sm28-GST, induced type 2, mixed, or type 1 immune responses, respectively. In the present study we examined whether the genetic background, the dose and the route of antigen administration could modulate the profile of the immune response induced during these immunizations. Our results show that the nature of the adjuvant is the major factor that determines the profile of the response. Surprisingly, the genetic background did not influence the response, while the route of immunization, and to a lesser extent the dose of the antigen, weakly modulated the adjuvant-dependent orientation of the immune response.

Adjuvants, Immunologic↗

PubMed1998

Human IgE responses to rSm22.6 are associated with infection intensity rather than age per se, in a recently established focus of Schistomiasis mansoni.

In studies of schistosomasis mansoni-endemic communities, individuals with IgE responses to a 22 kD adult worm antigen (rSm22.6) suffered lower intensities of reinfection after treatment. It is of interest to define the factors that lead to the production of rSm22.6-specific IgE because it is a marker for resistant individuals and it may be involved in the development of resistance to reinfection. In endemic populations rSm22.6-specific IgE increases linearly with age. However, it is not possible to distinguish between age per se and 'history of infection' in endemic populations because individuals are exposed to the parasite at an early age. We have, therefore, quantified pre- and post-treatment isotype responses to rSm22.6 in a comparatively 'epidemic' Senegalese community where the patients were infected at different ages and where pre-treatment intensity of infection can be taken as a reasonable measure of antigen exposure. Post-treatment isotype responses to rSm22.6 correlated positively with pre-treatment intensities of infection but were not shown to be related to age. IgG1, IgG4 and IgE responses to rSm22.6 were significantly higher after treatment with the difference increasing with the pre-treatment level of infection. These results from a recently established focus of infection suggest that isotype responses to rSm22.6 are antigen-exposure dependent rather than dependent on age per se.

Adolescent↗

PubMed1998

Molecular modeling of HIV-1 coreceptor CCR5 and exploring of conformational space of its extracellular domain in molecular dynamics simulation.

The chemokine receptor CCR5 functions as a major fusion coreceptor for macrophage-tropic human immunodeficiency virus entry into cell. Here we report a three-dimensional model of CCR5 built using molecular modeling approach. Because the virus binds to extracellular domain of the receptor, special attention was given to conformational flexibility, hydrogen bonding, and environmental polarity properties of this protein part. Such data were obtained in the result of molecular dynamics study of the extracellular domain. It was shown that during the simulation the extracellular segments form a compact globular domain with numerous long-range hydrogen bonds between them. First loop of the receptor stays quite rigid while N-terminal region and loops 2, 3 are rather flexible. A number of amino acid residues disposed in unfavourable environment and, therefore, potentially involved in binding of CCR5 to viral glycoproteins and chemokines, was delineated. Comparison of the results with available experimental data permits a proposal that such residues in loop-1 and N-terminal part of the receptor are important for HIV-1 entry, while those in loops 2 and 3 participate in ligand binding. Perspectives of rational alteration of virus-binding activity of CCR5 are discussed.

Amino Acid Sequence↗

PubMed1998

Systemic and mucosal immune responses after intranasal administration of recombinant Mycobacterium bovis bacillus Calmette-Guérin expressing glutathione S-transferase from Schistosoma haematobium.

A major goal of current vaccine development is the induction of strong immune responses against protective antigens delivered by mucosal routes. One of the most promising approaches in that respect relies on the use of live recombinant vaccine carriers. In this study, Mycobacterium bovis BCG was engineered to produce an intracellular glutathione S-transferase from Schistosoma haematobium (Sh28GST). The gene encoding Sh28GST was placed under the control of the mycobacterial hsp60 promoter on a replicative shuttle plasmid containing a mercury resistance operon as the only selectable marker. The recombinant Sh28GST produced in BCG bound glutathione and expressed enzymatic activity, indicating that its active site was properly folded. Both intraperitoneal and intranasal immunizations of BALB/c mice with the recombinant BCG resulted in strong anti-Sh28GST antibody responses, which were enhanced by a boost. Mice immunized intranasally produced a mixed response with the production of Sh28GST-specific immunoglobulin G1 (IgG1), IgG2a, IgG2b, and IgA in the serum. In addition, high levels of anti-Sh28GST IgA were also found in the bronchoalveolar lavage fluids, demonstrating that intranasal delivery of the recombinant BCG was able to induce long-lasting secretory and systemic immune responses to antigens expressed intracellularly. Surprisingly, intranasal immunization with the BCG producing the Sh28GST induced a much stronger specific humoral response than intranasal immunization with BCG producing the glutathione S-transferase from Schistosoma mansoni, although the two antigens have over 90% identity. This difference was not observed after intraperitoneal administration.

Administration, Intranasal↗

PubMed1998

Relationship of impairment of schistosome 28-kilodalton glutathione S-transferase (GST) activity to expression of immunity to Schistosoma mattheei in calves vaccinated with recombinant Schistosoma bovis 28-kilodalton GST.

Sera from calves vaccinated with the recombinant Schistosoma bovis-derived 28-kDa glutathione S-transferase (28GST) and subsequently naturally or experimentally exposed to Schistosoma mattheei were studied for their content of specific immunoglobulin G (IgG) and IgA antibodies to recombinant S. bovis 28GST as well as for their capacity to inhibit the enzymatic activity of the antigen. The results were analyzed in regard to the presence (natural infection) or absence (experimental infection) of a protective effect(s) (reductions in worm burden, egg load, fecal egg counts, and excretion of viable eggs) toward S. mattheei challenge. Under such conditions, no differences in the IgG- and IgA-specific antibodies to recombinant S. bovis 28GST or in the ability to block the catalytic function of the antigen between the two groups were recorded. Nevertheless, correlation analysis between the specific antibody responses to recombinant S. bovis 28GST and the inhibition of GST activity suggested an association with IgG in experimentally infected vaccinated animals, while in naturally infected vaccinated calves, the inhibitory activity appeared to be linked to a greater degree with IgA. These results suggest that in contrast to schistosomiasis in humans, IgG antibodies in calves with schistosomiasis may exhibit inhibitory functions toward GST enzymatic activity or have a modulatory effect on IgA antibody properties. Furthermore, sera from animals immunized with recombinant S. bovis 28GST recognized the native S. mattheei 28GST and achieved comparable levels of inhibition of activity of recombinant S. bovis 28GST and S. matthei 28GST, indicating the presence of cross-reactive epitopes on these two molecules.

Animals↗

PubMed1998

Bordetella pertussis filamentous hemagglutinin enhances the immunogenicity of liposome-delivered antigen administered intranasally.

In an attempt to increase the immunogenicity of mucosally delivered antigens, we incorporated the Bordetella pertussis filamentous hemagglutinin (FHA) adhesin into liposomes containing the glutathione S-transferase of Schistosoma mansoni (Sm28GST) as a model antigen. Outbred mice immunized twice intranasally with liposomes containing a constant suboptimal dose of Sm28GST and increasing doses of FHA produced anti-Sm28GST antibodies in a FHA dose-dependent manner. The addition of 3 microg of FHA to the liposomes induced more than 10-fold-higher anti-Sm28GST antibody titers, compared to those induced by liposomes without FHA. The presence of FHA did not alter the nature of the humoral immune response, and the sera contained anti-Sm28GST immunoglobulin G1 (IgG1), IgG2a, and IgG2b. However, anti-Sm28GST IgA was only detected when at least 3 microg of FHA was added to the preparation. These results show a promising potential for FHA to enhance the immunogenicity of mucosally administered antigens incorporated into liposomes.

Adhesins, Bacterial↗

PubMed1998

Inhibitory effects of lodoxamide on eosinophil activation.

Recent reports describe the beneficial use of lodoxamide, an anti-allergic compound, for the treatment of asthma and allergic conjunctivitis. Lodoxamide is known as a mast cell stabilizer, however, the association of a significant clinical improvement with a specific decrease in eosinophil infiltrate suggested possible direct effects of lodoxamide on eosinophils. The chemotactic response of eosinophils to fMLP as well as to IL-5, in vitro, was very significantly and dose-dependently inhibited by Lodoxamide. Lodoxamide was also able to strongly inhibit the release of eosinophil peroxidase after IgA-dependent activation and, to a lesser extent, the release of eosinophil cationic protein and eosinophil-derived neurotoxin. Moreover, the release of cytotoxic mediators evaluated in an antibody-dependent cytotoxicity assay against parasitic targets was also significantly reduced, not only in the case of human eosinophils but also in a rat eosinophil-mast cell model of cytotoxicity. Taken together, these results indicate that lodoxamide can exert potent inhibitory effects on eosinophil activation in vitro combined with a strong inhibition of eosinophil attraction, leading therefore to a reduction in their pathological potential in vivo.

Animals↗

PubMed1998

Glutathione S-transferases of 28kDa as major vaccine candidates against schistosomiasis.

For the development of vaccine strategies to generate efficient protection against chronic infections such as parasitic diseases, and more precisely schistosomiasis, controlling pathology could be more relevant than controlling the infection itself. Such strategies, motivated by the need for a cost-effective complement to existing control measures, should focus on parasite molecules involved in fecundity, because in metazoan parasite infections pathology is usually linked to the output of viable eggs. In numerous animal models, vaccination with glutathione S-transferases of 28kDa has been shown to generate an immune response strongly limiting the worm fecundity, in addition to the reduction of the parasite burden. Recent data on acquired immunity directed to 28GST in infected human populations, and new development to draw adapted vaccine formulations, are presented.

Animals↗

PubMed1998

Molecular cloning and sequencing of the rat interleukin-12 p40 gene.

The nucleotide sequences containing the rat interleukin-12 p40 gene was determined. Sequencing revealed the presence of six exons and five introns. Analysis of the 5' non-coding region showed the presence of several possible sites involved in cytokine gene regulation at the transcriptional level. Comparison of the deduced amino acid sequence of rat IL-12 p40 with that of the mouse and of human p40, showed 92% and 65% identity respectively.

Amino Acid Sequence↗

PubMed1998

Murine immune responses to Schistosoma haematobium and the vaccine candidate rSh28GST.

Longitudinal studies of Schistosoma haematobium infection in CBA mice revealed a progressive down-regulation of cellular immune responses, as measured by mitogenic and antigenic stimulation of in vitro lymphocyte cultures. Antigen-stimulated production of the Th1 cytokine IFN-gamma by splenocytes increased progressively up to 14 weeks post infection, (four weeks after the onset of parasite egg production), before declining swiftly. Levels of the Th2 cytokine IL-4 in the same cultures remained low until 14 weeks, after which they rose rapidly as IFN-gamma declined. High levels of IL-10 coincided with the peak in IFN-gamma production, suggesting a non Th2-restricted role for this cytokine. Both total and antigen-specific immunoglobulin production confirmed parasite egg deposition as being a major stimulus for host humoral responses. The S. haematobium infection failed to elicit detectable T cell responses to the antifecundity vaccine candidate rSh28GST. However, low levels of antibody were detectable in infected mouse serum and strong IgG and IgA production was induced by vaccination with rSh28GST plus adjuvant.

Animals↗