Biomedical subjects

H Crespeau

Publications and source records attributed to H Crespeau.

11 recordsLinked to original sources

PubMed2001

Homology between a 173-kb region from mouse chromosome 10, telomeric to the Ifng locus, and human chromosome 12q15.

We sequenced a 173-kb region of mouse chromosome 10, telomeric to the Ifng locus, and compared it with the human homologous sequence located on chromosome 12q15 using various sequence analysis programs. This region has a low density of genes: one gene was detected in the mouse and the human sequences and a second gene was detected only in the human sequence. The mouse gene and its human orthologue, which are expressed in the immune system at a low level, produce a noncoding mRNA. Nonexpressed sequences show a higher degree of conservation than exons in this genomic region. At least three of these conserved sequences are also conserved in a third mammalian species (sheep or cow).

Animals↗

PubMed1995

Flow cytometric monitoring of allergen induced basophil activation.

Human basophils express many clustered differentiation antigens (CD), including CD45; however, none are specific for these cells. In a previous study, we described a two-color immunofluorescence procedure, employing antibodies to CD45 and IgE for the cytometric evaluation of basophils. In the present work, we show that when sensitized basophils are stimulated by allergenic preparations, they demonstrate an upregulation of CD45 as well as a decrease in anti-IgE binding. Since CD45 antigen modulation was observed with all aeroallergens tested and the decrease in IgE expression varied with allergenic preparations, the measurement of CD45 upregulation was used to evaluate basophil activation. Using this approach, reproducible results were observed when atopic patients were tested at different time intervals. In addition, we show that the upregulation of CD45 on allergen stimulated basophils is a very rapid phenomenon that is observed after a few minutes and that this rapid flow cytometric method can also be used for investigations on nonallergenic mediators of basophil stimulation such as calcium ionophores.

Adult↗

PubMed1994

[Failure mechanisms in the transfusion process. Importance of anticipatory operational safety analysis].

The methods used for the safety previsional analysis of operations represent an interesting set of tools to follow the so-called transfusion process, defined as all the steps from donors sensitization to recipients follow-up. FMECA (Failure Mode Effects and Criticality Analysis) can be used as a prevention tool, independently of any dysfunction in the process. Of course, it can also be used following a failure, in order to analyse its causes and to apply specific corrections. Operation safety, quality insurance, epidemiologic surveillance and safety monitoring act in synergy. These three aspects of transfusion safety constitute a dynamic system.

Blood Banks↗

PubMed1994

[Polyclonal DNase abzyme produced by anti-idiotypic internal image method].

The concept of antigen internal image was applied to the production of catalytic antibodies. An antibody raised in rabbits to DNase (Ab1), acted as a competitive inhibitor of the catalysis, and thus was assumed to contain anti-active site Ab. This Ab1 was used to elicit a polyclonal anti-idiotypic antibody (Ab2). This later exhibited a DNA recognition specificity, suggesting the existence of structural internal images mimicking the conformation of the active site. Moreover, Ab2 were able to hydrolyse DNA, indicating the existence of internal images mimicking the enzymatic activity of DNase. Consequently, a strategy can thus be considered using the idiotypic way, for the production of abzymes in the form of internal images of enzyme active sites.

Antibodies↗

PubMed1992

Two alloalbumins with identical electrophoretic mobility are produced by differently charged amino acid substitutions.

We describe the amino acid substitutions of albumins Sondrio and Paris 2, two slow moving variants of human serum albumin, which show an identical electrophoretic mobility on cellulose acetate at three different pH values. These variants have been found in several instances in a wide geographic area including Northern Italy and France. Both alloalbumins were isolated from the sera of heterozygous subjects. Isoelectric focusing analysis of CNBr fragments from the purified variants allowed us to localize the mutation of albumin Sondrio in fragment CNBr V (residues 330-446) and that of albumin Paris 2 in CNBr VII (residues 549-585). Sequential analysis of the variant CNBr VII established the molecular defect of albumin Paris 2 as 563 Asp----Asn. Fragments CNBr V from normal and Sondrio albumins were isolated on a preparative scale and subjected to tryptic and V8 proteinase digestion. Sequence determination of the abnormal tryptic and V8 peptides revealed that the variant arises from the substitution of glutamic acid 333 by lysine. Thus, a +1 change in the C-terminal region of the albumin molecule produces a variant with the same electrophoretic mobility as an alloalbumin with a +2 substitution in the central domain, suggesting a higher degree of exposure to the solvent of the C-terminal tailpiece. Both amino acid substitutions are consistent with a G----A transition in the first position of the corresponding codon in the structural gene.

Amino Acid Sequence↗

PubMed1991

Albumin Paris 2: a new genetic variant distinguished by isoelectric focusing.

Until recently, the characterization of genetic variants of human serum albumin was performed by electrophoretic typing prior to the determination of their amino acid substitutions. We describe a procedure using isoelectric focusing in the presence of urea for the analysis of the genetic variation of albumin. This procedure allowed a clear distinction of a new variant, previously found to be identical with albumin Sondrio according to its relative electrophoretic mobilities at 3 pHs. This new variant, the third rare albumin allotype identified in the Ile-de-France region, was called albumin Paris 2.

Amino Acid Sequence↗

PubMed1991

[Characterization of genetic variants of human albumin by isoelectric focusing].

Normal human serum albumin and bisalbuminic fractions from genetic variants of european origin have been studied by ultrathin-layer isoelectric focusing performed on whole sera or after purification of albumin fractions by affinity chromatography on Blue-Trisacryl. Narrow range ampholytes giving pH gradient between 5 and 8, together with the use of 8 M urea, provided suitable patterns allowing to discriminate the various allotypes. In these conditions, both normal albumin and heterozygous variants were microheterogeneous with several main bands; in the case of normal albumin, four major bands were found, while variants exhibited additional bands differing in number and pI. The position of additional bands comparatively to that of normal albumin was consistent with the electrophoretic behavior of variants. Fast moving allotypes exhibited additional bands with more anodal pIs, whereas slow moving variants were characterized by cathodal additional bands. The number and position of these bands allowed to characterize some variants, when they failed to distinguish between some others, identical patterns being correlated to identical mutations arising at different locations on the albumin molecule. These observations indicate that isoelectric focusing could allow a clear distinction of albumin variants, provided that their mutation were different, or could confirm the occurrence of identical mutation when IEF patterns are indistinguishable. In addition to electrophoretic mobilities at various pH, analytical isoelectric focusing could be a useful technique employed as a second step in the identification of allotypes prior to the determination of the structural change characterizing the variant.

Blotting, Western↗

PubMed1991

[Diversity and relative inaccuracy of internal images of antigen group A revealed by IEF analysis].

The diversity of a polyclonal anti-idiotypic response (Ab2) to a murine monoclonal anti-A (Ab1) was investigated after purification of two Ab2 populations. One was eluted from human polyclonal anti-A column and the other from Ab1. Analysis of the Ab specificity, as well as screening of the clonotypic distribution, were achieved after splitting Ab by IEF; this was followed by immunoblotting and probing with various anti-ABH mAb. The first population reacted with almost all the murine anti-ABH mAb, as well as with four human anti-A mAb, and consequently consisted of Ab2 beta. The second was composed of "true" Ab2 directed against Ab1. In the first population internal images mimicked either A Ag, or H Ag, or some epitopes common to both. This study demonstrates the plurality of internal images-bearing Ig molecules, some mimicking completely, and some only partially or even unfaithfully the nominal A determinant. The analysis of this idiotypic cascade proves the existence of a degeneracy of the initial restricted antigenic specificity. The consequences of such a process are discussed.

Animals↗

PubMed1989

A sensitive double-diffusion microassay suitable for the detection of idiotype-antiidiotype precipitates.

Using agarose gel coated on GelBond film sheets and using Coomassie blue stain followed by silver stain, a sensitive double-diffusion microassay has been developed for detecting small amounts of precipitate forming during idiotype-antiidiotype reactions. The sensitivity of the method is 10-100 times greater than classical immunodiffusion tests. Other advantages include the need for minimal amounts of unconcentrated sample, the absence of radioactive or toxic substrates, no interference due to a second or third antibody coat such as are used in immunoenzymatic techniques, and the possibility of a direct evaluation of qualitative data such as identity, cross-reactivity or non-identity. As little as 40 ng antibody could be detected, corresponding to an antibody concentration of 8 micrograms/ml, making the microassay useful for rapid screening of idiotype-antiidiotype precipitates during routine analysis of hybridoma supernatants.

ABO Blood-Group System↗

PubMed1989

[Detection and quantification of weak concentrations of antigens using a sensitive and direct method of demonstrating immunoprecipitation reactions].

Using agarose gel coated on GelBond film sheets, and using Coomassie blue stain followed by silver stain, a sensitive microassay has been developed for detecting small amounts of antigen-antibody precipitates, and for quantitating low concentrations of antigen. In order to obtain a high sensitivity, antigen-antibody ratios were adjusted imperatively close to the equivalence in double-diffusion, and for quantitative estimation, single radial immunodiffusion was performed, according to Mancini, by measuring circles at the end point. The use of a double staining procedure allows to detect as little as 8 micrograms/ml antigen by Ouchterlony and 200 ng/ml by Mancini technique. The sensitivity of the method is 100 times greater than classical techniques and other advantages such as the need for minimal amounts of unconcentrated samples, the absence of radioactive labelling, and the absence of interference due to a second or a third antibody coat, make this assay useful for analyzing and quantitating monoclonal antibodies obtained by hybridoma or B-cell immortalization.

Antibodies, Monoclonal↗

PubMedDate not supplied

ABO-blood-group-related idiotypic network: mimicry of oligosaccharide epitope by rabbit antiidiotypic antibodies to murine monoclonal anti-A antibody.

The idiotypy of antibodies (Ab) specific for oligosaccharide determinants of blood groups of the human ABO system was studied through a cascade. Xenogenic antiidiotypic Ab (Ab2) raised in rabbits to the murine monoclonal anti-A61 (Ab1) were screened for reactivity with various anti-ABH Ab. Three anti-A and three anti-A,B monoclonal antibodies (mAb) which were developed in the same mouse strain as that producing Ab1, as well as a human polyclonal anti-A, were found to share cross-reactive idiotopes (CRI) with Ab1. CRI on murine mAb could be due to a Biozzi recurrent Id on anti-A Ab reacting with anti-Id "à la Oudin", while CRI on human anti-A Ab suggested the presence of paratope-induced anti-Id. Inhibition by Ab2 of haemagglutination of A, B or O human red blood cells by many murine anti-ABH mAb, and by polyclonal or monoclonal human anti-A, strongly supported the occurrence of anti-Id mimicking ABH epitopes belonging to type 2 determinants carried by human erythrocytes. Furthermore, a rabbit immunized with Ab2 produced a potent Ab3 response characterized by anti-H-type-2 specificity. Altogether, these results are consistent with the first successful production of anti-Id Ab that mimics the tridimensional shape of a well defined and strictly carbohydrate epitope, eliciting a haemagglutinating Ab3.

ABO Blood-Group System↗