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A Capron

Publications and source records attributed to A Capron.

At least 127 records · Page 7Linked to original sources
PubMed1995

Eosinophils: from low- to high-affinity immunoglobulin E receptors.

Several experimental approaches have been used to identify immunoglobulin (IgE) binding molecules expressed by human eosinophils. After the description that Fc epsilon RII/CD23 identified on eosinophils could participate in IgE binding and IgE-mediated cytotoxicity, Mac2/epsilon binding proteins belonging to the S-type lectin family were also detected on human eosinophils. Anti-Mac2 monoclonal antibodies inhibited eosinophil-dependent cytotoxicity towards parasitic targets. More recently, Fc epsilon RI was demonstrated on human eosinophils from hypereosinophilic patients. The 3 components of Fc epsilon RI, alpha, beta and gamma chains, were detected in eosinophils. The alpha chain of Fc epsilon RI was shown to be involved in IgE binding to eosinophils and in the selective release of eosinophil peroxidase. The participation of Fc epsilon RI-bearing eosinophils in a protective immune response against a parasitic infection indicates a so far unsuspected function of Fc epsilon RI. The interactions between the different types of IgE binding molecules are discussed.

Antigens, Differentiation↗
PubMed1995

Analysis of the Mycobacterium tuberculosis 85A antigen promoter region.

A mycobacterial expression-secretion vector was constructed in which the Escherichia coli alkaline phosphatase (phoA) reporter gene was placed under the control of the Mycobacterium tuberculosis 85A promoter and secretion signal sequences. In recombinant Mycobacterium smegmatis and Mycobacterium bovis BCG, PhoA activity could readily be detected on the mycobacterial cell surface and in the culture supernatant, indicating that the 85A signals can drive heterologous expression and secretion in both species. In contrast to the mycobacteria, the 85A promoter did not function in E. coli. We mapped the promoter region by progressive deletions using BAL 31 exonuclease and by primer extension analysis. Insertion and deletion mutations within the promoter region indicated that, unlike most E. coli promoters but similar to Streptomyces promoters, the position of the putative -35 region was not critical for efficient promoter activity. In addition, we investigated the ability of the identified signals to drive the production and secretion in BCG of recombinant Schistosoma mansoni glutathione S-transferase (Sm28GST), a protective antigen against schistosomiasis. BALB/c mice immunized with the recombinant BCG by a single dose exhibited a weak but specific T-cell response to Sm28GST.

Alkaline Phosphatase↗
PubMed1994

Construction, expression, and immunogenicity of multiple tandem copies of the Schistosoma mansoni peptide 115-131 of the P28 glutathione S-transferase expressed as C-terminal fusions to tetanus toxin fragment C in a live aro-attenuated vaccine strain of Salmonella.

Genetic fusions have been constructed between the highly immunogenic but atoxic fragment C of tetanus toxin and a guest peptide, aa115-131, from the protective 28-kDa glutathione S-transferase Ag of Schistosoma mansoni. Fusions have been assembled to express one, two, four, and eight tandem copies of the peptide. The recombinant vectors have been electroporated into the nonvirulent aroA strain of Salmonella typhimurium SL3261. The fusion proteins are soluble and stably expressed in Salmonella as evaluated by Western blotting with fragment C and glutathione S-transferase antisera. Mice have been immunized i.v. with a single dose of the live recombinant salmonellae. The strains are stable in mice and elicit Ab responses directed against fragment C, as determined by enzyme-linked immunosorbent assays. Ab responses were also detected against the guest peptide. The Ab responses improved dramatically toward the aa115-131 peptide with increasing copy number, with the octameric "repitope" fusion displaying the greatest potency. This approach may represent a general strategy for eliciting immune responses against peptides in live bacterial vaccines.

Amino Acid Sequence↗
PubMed1994

Construction, expression, and immunogenicity of the Schistosoma mansoni P28 glutathione S-transferase as a genetic fusion to tetanus toxin fragment C in a live Aro attenuated vaccine strain of Salmonella.

A vector has been constructed to allow genetic fusions of guest antigens via a hinge domain to the C terminus of the highly immunogenic C fragment of tetanus toxin. A fusion has been constructed with the gene encoding the protective 28-kDa glutathione S-transferase (EC 2.5.1.18) from Schistosoma mansoni. The recombinant vector has been electroporated into the nonvirulent Salmonella typhimurium aroA live vaccine strain SL3261. The corresponding chimeric protein is stably expressed in a soluble form in Salmonella as evaluated by Western blotting with fragment C and glutathione S-transferase antisera. Mice immunized intravenously with a single dose of the live recombinant bacteria elicit antibodies to both fragment C and glutathione S-transferase as detected by enzyme-linked immunosorbent assays. Furthermore, all of the mice were solidly protected when challenged with lethal doses of either tetanus toxin or the virulent Salmonella typhimurium strain C5. Mice have also elicited antibodies to fragment C and glutathione S-transferase after oral immunization. It may be that a live trivalent vaccine against typhoid, tetanus, and schistosomiasis is feasible.

Animals↗
PubMed1994

Human IgE in SCID mice reconstituted with peripheral blood mononuclear cells from Dermatophagoides pteronyssinus-sensitive patients.

SCID mice were reconstituted with purified peripheral blood mononuclear cells from patients sensitive to Dermatophagoides pteronyssinus (Dpt). After immunization with a low dose of the related allergen, these human (Hu)-SCID mice may develop a specific IgE response. By using an IgE-dependent platelet assay and the related allergen Dpt, the human IgE was demonstrated to be functional. Indeed a high correlation was obtained between both parameters. No rise in human IgE level was detected within 3 wk after the immunization with an unrelated Ag. When mice were reconstituted with peripheral blood cells from nonallergic donors and immunized with the Dpt allergen, no human IgE production was evidenced in the serum of SCID mice. However, in both allergic Hu-SCID mice (reconstituted with cells from allergic patients) or in nonallergic Hu-SCID mice (reconstituted with cells from healthy donors), an IgG response was detectable. Finally by radioallergosorbent test, a specific IgE Ab response was detected after the first allergen challenge only in allergic Hu-SCID mice. Thus, without ethical constraints, SCID mice represent a useful model for analyzing the IgE response occurring in allergic patients.

Allergens↗
PubMed1994

A radioimmunoassay for the quantification of human ubiquitin in biological fluids: application to parasitic and allergic diseases.

Purified ubiquitin has been shown to share similar biological and physicochemical properties with a previously characterized lymphokine, platelet activity suppressive lymphokine (PASL). This lymphokine, which inhibits the cytotoxic function of activated platelets, is produced during schistosomiasis and Hymenoptera venom hypersensitivity (HVH). We have developed a radioimmunoassay specific for ubiquitin, in order to determine the ubiquitin levels in human sera and plasma from patients with these pathologies. The working range of the assay was between 60 and 500 ng/ml, and the sensitivity was 8-10 ng/ml. The reproducibility, specificity and accuracy were determined under standard condition (PBS-0.3% BSA) and using different biological fluids (human serum, plasma and T lymphocyte supernatant). Using this assay, we found that the ubiquitin concentrations were higher in both schistosomiasis and HVH (up to 150-300 ng/ml) compared with sera and plasma from healthy donors where the ubiquitin levels did not exceed 50 ng/ml.

Anaphylaxis↗
PubMed1994

Similarities between the primary structures of two distinct major surface proteins of Toxoplasma gondii.

Toxoplasma gondii possesses a 43-kDa surface protein (SAG3) that is expressed by all invasive stages. We have cloned and sequenced cDNAs encoding SAG3, with the longest one encoding a primary product of 385 amino acid residues. The deduced amino acid sequence contains a putative NH2-terminal signal sequence, as well as a glycosylphosphatidylinositol anchor attachment site. It is characterized by 12 cysteine residues whose distribution suggests a tandem duplication of a single ancestral motif containing 6 cysteine residues. Although no DNA sequence analogies were found, comparative amino acid sequence analysis detected a resemblance to SAG1, which is the major surface antigen specifically expressed by the proliferative tachyzoite stage. Despite a low degree of identity between the two amino acid sequences (24%), the conservative distribution of the cysteine and tryptophan residues, as well as of repeated motifs, together with oligopeptide identities suggest similar folding and possibly similar function for both proteins.

Amino Acid Sequence↗
PubMed1994

Analysis of the V beta specificity of superantigen activation with a rapid and sensitive method using RT PCR and an automatic DNA analyser.

Polymerase chain reaction (PCR) by the specific amplification of a DNA target sequence has been shown to permit analysis of T cell receptor usage. The complete repertoire is amplified using oligonucleotide primers specific for each of the known V alpha or V beta regions of the T cell receptor. One of the methods currently used to appreciate the relative quantity of different V chains of the TCR is by coamplifying in the same reaction tube the variable region of one chain together with the constant region of the other chain. We have optimised PCR conditions and analysed PCR products on an automatic DNA analyser facilitating the quantification of the amplified products, avoiding the use of radioisotopes, and allowing the determination of the sizes of CDR3 regions, thus giving new information on the modification of the T cell repertoire. This method was used to analyse the precise V beta specificity of the T cell activation with the superantigen SEB.

Autoanalysis↗
PubMed1994

Interleukin 5 synthesis by eosinophils: association with granules and immunoglobulin-dependent secretion.

Interleukin 5 (IL-5) is the main factor that promotes the terminal differentiation of eosinophil progenitors (as indicated by colony formation assays), and enhances the effector capacity of mature eosinophils. IL-5 is produced by T lymphocytes, CD4-/CD8- and mast cells and recently, messenger (m)RNA of this cytokine has been identified in eosinophils from patients with coeliac disease, asthma, or eosinophilic heart diseases. In this study, IL-5 mRNA and immunoreactive IL-5 protein were detected in tissue and blood eosinophils from patients with eosinophilic cystitis or hypereosinophilic syndromes but not in Crohn's disease. By electron microscopy associated to immunogold staining, immunoreactive IL-5 was identified in eosinophilic granules. After stimulation with IgA-, IgE-, or IgG-immune complexes, blood eosinophils were shown, by immunocytochemistry and by enzyme-linked immunosorbent assay, to secrete IL-5. These observations demonstrate that eosinophils, under physiological stimulation, can release significant amounts of IL-5, which may contribute to local eosinophil recruitment and activation.

Cytoplasmic Granules↗
PubMed1994

Cloning and characterization of the gene encoding Schistosoma mansoni glutathione peroxidase.

Antioxidant enzymes are thought to play a crucial role in the survival of the parasite, Schistosoma mansoni, during its migration through the tissues of the definitive host. We recently cloned the cDNA encoding one such enzyme, glutathione peroxidase (Gpx). In order to elucidate the regulation of expression of this gene, we describe the cloning and characterization of a Gpx gene of S. mansoni. An initial screen of a lambda EMBL4 genomic library using the corresponding cDNA sequence as a probe yielded 14 positive clones, two of which have so far been analyzed in detail. The complete Gpx gene contains five introns, four of which, located at the 5' end, are extremely short (30-51 bp) and the last of which is approximately 6 kb long. We present the sequence of the gene including 73 bp at the 5' end, the complete sequence to 137 bp downstream from the penultimate exon, 164 bp upstream and 131 bp downstream from the last 3' exon. The potential mRNA cap site is situated 219 bp upstream from the ATG start codon. All intron/exon junctions correspond to the conventional eukaryotic splice signal. Analysis of the 5' flanking region revealed the presence of a potential TATA box at--26 bp from the cap site, but no CAAT-like element is present. Southern blot analysis showed a unique Gpx gene organisation in the S. mansoni genome.

Amino Acid Sequence↗
PubMed1994

High-affinity IgE receptor on eosinophils is involved in defence against parasites.

Parasitic infections are often associated with eosinophilia and high levels of immunoglobulin E (IgE). This observation has led to speculation that eosinophils and IgE may act together in the immune response against parasites. In support of this hypothesis, IgE and eosinophils participate in cytotoxic reactions directed against Schistosoma mansoni larvae in vitro. Furthermore, epidemiological studies have shown an inverse correlation between levels of specific IgE and rates of infection with Schistosoma. The low-affinity IgE receptor (Fc epsilon RII/CD23) was first incriminated in eosinophil activation. The fact that the high-affinity IgE receptor (Fc epsilon RI) is not only expressed on mast cells and basophils but also on Langerhans cells led us to investigate the presence of Fc epsilon RI on eosinophils. Here we show that Fc epsilon RI is expressed on eosinophils from hypereosinophilic patients, is involved in eosinophil degranulation, and participates in eosinophil-mediated cytotoxicity against S. mansoni. Our results indicate that Fc epsilon RI may play a major part in immune defence against parasites.

Animals↗
PubMed1994

The mixotope: a combinatorial peptide library as a T cell and B cell immunogen.

We report a new approach in peptide vaccine strategy based on combinatorial synthesis. A library of 7.5 x 10(5) related peptides, termed mixotope, was derived from the sequence of the third hypervariable domain (V3 loop) of the human immunodeficiency virus (HIV) envelope protein. This preparation induced a strong immune response in all syngeneic and outbred rodents tested. The response directed against the mixotope included antibodies, CD4+ T helper cells (TH1 and TH2) and CD8+ T cells. In rodents immunized with the mixotope, the T cell response directed against individual V3 peptide sequences (BRU, MN, RF, SF2, and ELI) as measured by T cell proliferation and interleukin (IL)-2 production, was found to be major histocompatibility complex haplotype-dependent. However, additional experiments performed in mice indicated that selectivity was less restrictive when using IL-3 secretion to explore T cell activation. This combinatorial antigen could be considered as a series of agretopic motifs framing a multiplicity of closely related epitopes for T cell recognition and able to elicit a T cell and B cell repertoire. This new construct may therefore provide a basis for the design of future vaccine strategies.

AIDS Vaccines↗