Search PubMed⌕ Search

Biomedical subjects

C Babinet

Publications and source records attributed to C Babinet.

At least 109 records · Page 6Linked to original sources
PubMed1987

Idiotypic relationships between allotypically different rabbit antibodies.

Homozygous rabbits genotypically different for the allotypy of the VH region and the C kappa region of their immunoglobulins (al/al, b4/b4 and a3/a3, b5/b5, respectively) were transferred together at the blastocyst stage to be born from the same foster mother. At the time of transfer the foster mothers were active anti-anti-idiotypic (Ab3) antibody producers and, according to their own genotype, the idiotypes (Ab1) which initiated the immunization cascade ending with their Ab3 were allotypically either a1,b4 or a3,b5. This experimental scheme was carried out, at the level of the Ab1 and of the Ab'1 (antibodies produced by the transferred rabbits subjected to the Ab3 action), with three different exogenous antigens, namely a protein, a polysaccharide and a hapten which elicited in normal rabbits the production of antibodies with private idiotypic specificities. Two of them, the protein and the polysaccharide enabled us to find idiotypic relationships between antibodies totally different for their allotypy and particularly for the allotypy of the variable region of their heavy chains. In these examples, the structural coercions implied by the different allotypic markers of the VH did not seem to prevent the association of at least related genetic elements responsible for the observed idiotypic community. In these idiotypic relationships, only subpopulations of the Ab1 and of the Ab'1 were implicated and they could be different for the diverse littermates.

Animals↗
PubMed1987

Rearrangement and expression of rabbit immunoglobulin kappa light chain gene in transgenic mice.

To determine whether a foreign unrearranged immunoglobulin gene can be functionally rearranged and expressed in vivo, a rabbit b9 kappa light chain gene construct containing a single germ-line kappa chain variable (V) region gene (V kappa), the five kappa chain joining (J) segments (J kappa), and the kappa chain constant (C) region germ-line gene (C kappa) was introduced into fertilized mouse eggs. Eleven transgenic mice carrying 1-30 copies of the rabbit kappa gene construct were obtained. Rearrangement of the transgene due to V kappa-J kappa recombination was observed in the spleen of all the mice lines analyzed. Only the J kappa 1 and J kappa 2 segments, which have canonical sequences for rearrangement and high-level expression, are utilized in assembly of the rabbit gene. V kappa-J kappa 1 and V kappa-J kappa 2 joining was also observed in the thymus but not in nonlymphoid tissue. Polyadenylylated rabbit kappa chain transcripts of 1.2 kilobases were found in the spleen of the transgenic mice. The level of transcription was low despite a high level of rearrangement. Three transgenic mice lines secreted kappa light chains encoded by the foreign rabbit gene. Serum rabbit kappa chains were associated with mouse mu and gamma 1 heavy chains. However, hybrid antibody molecules containing both rabbit and mouse kappa light chains were also found in the serum of these animals. These results suggest that, although the rabbit kappa chain gene construct contains the necessary sequences for gene assembly, sequences implicated in stage- and tissue-specific regulation of kappa chain gene rearrangement are either missing or not recognized by mouse lymphoid cells.

Animals↗
PubMed1987

Hepatitis B surface antigen gene expression is regulated by sex steroids and glucocorticoids in transgenic mice.

We have investigated the basis for liver-specific and sex-linked expression of hepatitis B surface antigen (HBsAg) gene in transgenic mice by monitoring the level of liver HBsAg mRNA and serum HBsAg at different stages of development and in response to sex-hormone regulation. Transcription of the HBsAg gene starts at day 15 of development, together with that of the albumin gene, and reaches a comparable level at birth. HBsAg mRNA level and HBsAg production are parallel in males and females during prenatal development and until the first month of life, but HBsAg gene expression increases 5-10 times in males at puberty. After castration, the level of expression decreases dramatically in both males and females and is subsequently increased by injection of testosterone or estradiol. Glucocorticoids also regulated positively expression of the HBsAg gene. Our results suggest that sex hormones play a role in hepatitis B virus gene expression during natural infection and could explain the difference in incidence of chronic carriers between men and women.

Animals↗
PubMed1986

Transhybridomas from fusion gene transgenic lymphocytes can be used to produce foreign protein of biological interest.

A method is described for achieving the efficient production of proteins of biological interest by the establishment of hybridomas from lymphocytes of transgenic animals carrying a fusion gene having promoter-regulatory sequences functional in lymphocytes fused to the coding sequences of the protein to be produced. While possible improvements in the method are discussed, it is anticipated that the method will ultimately be applicable to the production of any protein of interest.

Animals↗
PubMed1986

Studies on the expression of an H-2K/human growth hormone fusion gene in giant transgenic mice.

Transgenic mice carrying the H-2K/human growth hormone (hGH) fusion gene were produced by microinjecting into the pronucleus of fertilized eggs DNA molecules containing 2 kb of the 5' flanking sequences (including promoter) of the class I H-2Kb gene joined to the coding sequences of the hGH gene. Thirteen transgenic mice were obtained which all contained detectable levels of hGH hormone in their blood. Nine grew larger than their control litter-mates. Endogenous H-2Kb and exogenous hGH mRNA levels were analysed by S1 nuclease digestion experiments. hGH transcripts were found in all the tissues examined and the pattern of expression paralleled that of endogenous H-2K gene expression, being high in liver and lymphoid organs and low in muscle and brain. Thus 2 kb of the 5' promoter/regulatory region of the H-2K gene are sufficient to ensure regulated expression of hGH in transgenic mice. This promoter may therefore be of use to target the expression of different exogenous genes in most tissues of transgenic mice and to study the biological role of the corresponding proteins in different cellular environments.

Animals↗
PubMed1986

Identification of a paternal developmental effect on the cytoplasm of one-cell-stage mouse embryos.

Matings of female DDK mice with males of the BALB/c strain are sterile, whereas reciprocal crosses are normally fertile. We used nuclear transplantation between the hybrid eggs of these two strains to investigate the basis of this effect. We demonstrate that the observed sterility results from early embryonic mortality, that the mortality is due to a modification of the egg cytoplasm, and that the modification is mediated by the male pronucleus. Once established, this modification may affect female pronuclei of unrelated genotype such as C57BL/6. These results support the notion that a product derived from the male genome acts at the pronuclear stage and can affect later stages of embryonic development.

Animals↗
PubMed1986

Negative regulation of early polyomavirus expression in mouse embryonal carcinoma cells.

Embryonal carcinoma cells are resistant to infection by polyomavirus (Py). We showed that this block was partially removed by inhibiting protein synthesis temporarily. The block was also partially removed when Py was coinfected with simian virus 40. Cycloheximide treatment of cells infected with Py mutants able to grow on PCC4 embryonal carcinoma cells led to 3- to 10-fold increases in the production of T-antigen-positive cells. At 31 degrees C, Py T-antigen expression was enhanced when the cells were treated with cycloheximide. We suggest that a negative labile regulatory protein(s) is synthesized in PCC4 cells, preventing the initiation of early Py transcription by binding to the noncoding sequence, especially the enhancer element B and perhaps also element A, and that the Py mutants retained a binding site(s).

Animals↗
PubMed1985

Specific expression of hepatitis B surface antigen (HBsAg) in transgenic mice.

Two transgenic mice were obtained that contain in their chromosomes the complete hepatitis B virus (HBV) genome except for the core gene. These mice secrete particles of HBV surface antigen (HBsAg) in the serum. In one mouse, HBV DNA sequences that had integrated at two different sites were shown to segregate independently in the first filial generation (F1) and only one of the sequences allowed expression of the surface antigen. Among these animals the males produced five to ten times more HBsAg than the females. A 2.1-kilobase messenger RNA species comigrating with the major surface gene messenger RNA is expressed specifically in the liver in the two original mice. The results suggest that the HBV sequences introduced into the mice are able to confer a tissue-specific expression to the S gene. In addition, the HBV transgenic mice represent a new model for the chronic carrier state of hepatitis B virus infection.

Animals↗
PubMed1985

Asynchronous regulation of mouse H-2D and beta-2 microglobulin RNA transcripts.

The major transplantation (or H-2) antigens in the mouse are cell-surface glycoproteins composed of a heavy chain and a light chain, the beta-2 microglobulin (beta 2m). The expression of these proteins is regulated during development. Embryonic cells at early stages of development do not express these proteins. On the other hand, these molecules are present on the surface of all adult somatic cells. We investigated whether the expression of both chains was coordinately regulated. Using specific single-stranded DNA probes in an S1 nuclease analysis, we compared the relative amounts of H-2D and beta 2m transcripts in normal tissues, in transformed cells, and during embryonic development. Our results show that (1) the steady state level of beta 2m transcripts varies from one adult organ to another, while that of H-2D transcripts stays approximately the same; (2) upon transformation, the amount of H-2D-specific mRNA increases drastically, while the beta 2m mRNA level remains constant; (3) whereas the quantity of beta 2m mRNA increases during early development, the amount of H-2D mRNA remains at a very low level. These data suggest that the regulation of H-2D and beta 2m genes are not identical and that their activation during development is not synchronous.

Aging↗
PubMed1985

Expression of the cytokeratin endo A gene during early mouse embryogenesis.

Expression of cytokeratin endo A has been analyzed during mouse blastocyst formation and embryonal carcinoma cell differentiation. To study the regulation of endo A expression, nuclease S1 mapping experiments have been performed on RNA extracted from two-cell to 7.5-day embryos. Low levels of endo A mRNA begin to be detectable in eight-cell embryos. The amount of this mRNA increases at the blastocyst stage, suggesting that endo A expression is regulated at the mRNA level during blastocyst formation. At this stage, in situ hybridization studies show that endo A mRNA is present in the trophectoderm but not in the inner cell mass. In 7.5-day embryos, endo A mRNAs are also detectable in the endoderm layer and in the amnion.

Animals↗
PubMed1984

High survival of mouse embryos after rapid freezing and thawing inside plastic straws with 1-2 propanediol as cryoprotectant.

A method for obtaining a high survival rate of frozen-thawed mouse embryos is presented. Eight-cell mouse embryos were frozen inside small plastic straws in the presence of 1-2 propanediol and stored at -196 C. After thawing, the embryos were diluted for only 5 min in a 1.0 M sucrose solution to remove the 1-2 propanediol from the cells. At high rate of thawing (is equivalent to 2500 C/min) more than 88% of the embryos survived in vitro to the blastocyst stage provided that the dilution of propanediol was performed rapidly during thawing. At a lower rate of thawing (is equivalent to 300 C/min), survival tended to be higher (94.7%) when dilution was done 5 min after thawing. When the frozen-thawed embryos were transferred to the oviducts of day 1 pseudopregnant recipients either directly after the dilution of 1-2 propanediol or after 24 or 48 hr of culture, a high proportion of them (65.9%) develop normally to viable fetuses.

Animals↗
PubMed1984

Tetraparental rabbits chimeric for their lymphoid system--I. Allotype expression.

We constructed tetraparental rabbits by immunosurgically prepared inner-cell-mass transplantation. The two constitutive parental strains were chosen so that they not only differed by their coat colour (brown and white rabbits) but also by the allotypic specificities of their immunoglobulins. It was thus possible to distinguish among the rabbits born from composite blastocysts those which were chimeric for their lymphoid system and to unambiguously quantitate the respective contribution of each parental strain to this chimerism. In this peculiar situation, the possible perturbation of the expression of immunoglobulin allotypic specificities could be tested. Thus further analysis of the tetraparental rabbit's immunoglobulins was performed to look at possible anomalies of their constitution. In only one out of the 20 tetraparental rabbits studied, we found IgG having heavy chains with the allotypic specificities of one parental strain and light chains with the allotypic specificities of the other parental strain. We did not observe, in any case, the expression of immunoglobulin allotypic specificities other than the expected ones. We always observed a marked decrease with time in the expression of the immunoglobulins produced by the lymphocytes of one parental strain accompanied by a concomitant increase in the expression of the immunoglobulins produced by the lymphocytes of the other parental strain. Two tetraparental rabbits were found to be germ-line chimeras. Analysis of the immunoglobulin allotypic specificities of the progeny of our tetraparental rabbits including these two did not reveal any perturbation in their hereditary transmission according to the usual Mendelian laws.

Animals↗
PubMed1984

Tetraparental rabbits chimeric for their lymphoid system--II. Idiotype expression.

In order to test some aspects of the network theory of "idiotype-anti-idiotype" interactions, we studied the idiotypic specificities of anti-DNP antibodies produced by tetraparental rabbits, chimeric for their lymphoid system, under two different experimental conditions. In the first, the tetraparental rabbits were simply immunized against the hapten and we observed that the antibodies, totally different in their allotypy, produced by the lymphocytes of the two constitutive parental strains remained idiotypically independent in three cases out of three. In the second experimental regimen, the tetraparental rabbits underwent an anti-anti-idiotypic immunization before the anti-hapten one. In two out of the four cases studied we observed that anti-DNP antibodies different in their a- and b-series allotypic specificities can bear at least related idiotypic specificities.

Animals↗
PubMed1984

Altered expression of heat shock proteins in embryonal carcinoma and mouse early embryonic cells.

In a previous paper, we have shown that in the absence of stress, mouse embryonal carcinoma cells, like mouse early embryo multipotent cells, synthesize high levels of 89- and 70-kilodalton heat shock proteins (HSP)(O. Bensaude and M. Morange, EMBO J. 2:173-177, 1983). We report here the pattern of proteins synthesized after a short period of hyperthermia in various mouse embryonal carcinoma cell lines and early mouse embryo cells. Among the various cell lines tested, two of them, PCC4-Aza R1 and PCC7-S-1009, showed an unusual response in that stimulation of HSP synthesis was not observed in these cells after hyperthermia. However, inducibility of 68- and 105-kilodalton HSP can be restored in PCC7-S-1009 cells after in vitro differentiation triggered by retinoic acid. Similarly, in the early mouse embryo, hyperthermia does not induce the synthesis of nonconstitutive HSP at the eight-cell stage, but induction of the 68-kilodalton HSP does occur at the blastocyst stage. Such a transition in the expression of HSP has already been described for Drosophila melanogaster and sea urchin embryos and recently for mouse embryos. It may be a general property of early embryonic cells.

Animals↗
PubMed1981

Cell-cell interactions in early embryogenesis: a molecular approach to the role of calcium.

Compaction, a process of cell-cell adhesion between mouse blastomeres or between embryonal carcinoma (EC) cells requires calcium ions. A decompaction effect similar to that observed in the absence of Ca2+ is triggered by Fab fragments of rabbit anti-EC IgG. This effect occurs through the recognition of a specific cell-surface glycoprotein named uvomorulin. An 84,000 dalton fragment of uvomorulin (UMt) has been previously extracted by trypsin from EC cell membranes and purified. WE present evidence that effects of Ca2+ on compaction are transmitted through conformational changes in uvomorulin. First, Ca2+ protects UMt from further proteolysis by trypsin. Mn2+ and Sr2+ have similar effects, whereas this protection is reversed by La3+. Second, UMt can bind the monoclonal antibody De1 only in the presence of Ca2+ (half-binding at 10(-5) M Ca2+). This antigenic exposure also takes place in the presence of Mn2+ or Sr2+ and is reversed by La3+. Third, metal ions (Ca2+, Mn2+, Sr2+) that promote trypsin resistance and recognition by DE1 are found to trigger the compaction of morulae and EC cells. Metal ions (La3+) that reduce trypsin resistance and affinity for DE1 result in decompaction.

Animals↗
PubMed1980

Serological identification and cellular distribution of three F9 antigen components.

Using an affinity chromatography technique, IgM, IgG1, IgG2a,b anti-F9 antibodies have been isolated from the anti-F9 serum; their activities have been analyzed by IF test on a variety of cell types, teratocarcinoma-derived cell lines, and embryos. The anti-F9 antibodies react with at least three independent antigenic determinants not expressed on the same cell types, and that appear along different time-course during embryonic development.

Animals↗