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F Guillet

Publications and source records attributed to F Guillet.

At least 19 recordsLinked to original sources

Structure and diversity of Mexican axolotl lambda light chains.

We report here the structure of cDNA clones encoding axolotl light chains of the lambda type. A single IGLC gene and eight different potential IGLV genes belonging to four different families were detected. The axolotl Cgamma domain has several residues or stretches of residues that are typically conserved in mammalian, avian, and Xenopus Cgamma, but the KATLVCL stretch, which is well conserved in the Cgamma and T-cell receptor Cbeta domains of many vertebrate species, is not well conserved. All axolotl Vgamma sequences closely match several human and Xenopus Vgamma-like sequences and, although the axolotl Cgamma and Vgamma sequences are very like their tetrapod homologues, they are not closely related to nontetrapod L chains. Southern blot experiments suggested the presence of a single IGLC gene and of a limited number of IGLV genes, and analysis of IGLV-J junctions clearly indicated that at least three of the IGLJ segments can associate with IGLV1, IGLV2, or IGLV3 subgroup genes. The overall diversity of the axolotl Vgamma CDR3 junctions seems to be of the same order as that of mammalian Vgamma chains. However, a single IGLV4 segment was found among the 45 cDNAs analyzed. This suggests that the axolotl IGL locus may have a canonical tandem structure, like the mammalian IGK or IGH loci. Immunofluorescence, immunoblotting, and microsequencing experiments strongly suggested that most, if not all L chains are of the gamma type. This may explain in part the poor humoral response of the axolotl.

Ambystoma mexicanum↗

Conserved structure of amphibian T-cell antigen receptor beta chain.

All jawed vertebrates possess well-differentiated thymuses and elicit T-cell-like cell-mediated responses; however, no surface T-cell receptor (TCR) molecules or TCR genes have been identified in ectothermic vertebrate species. Here we describe cDNA clones from an amphibian species, Ambystoma mexicanum (the Mexican axolotl), that have sequences highly homologous to the avian and mammalian TCR beta chains. The cloned amphibian beta chain variable region (V beta) shares most of the structural characteristics with the more evolved vertebrate V beta and presents approximately 56% amino acid identities with the murine V beta 14 and human V beta 18 families. The two different cloned axolotl beta chain joining regions (J beta) were found to have conserved all the invariant mammalian J beta residues, and in addition, the presence of a conserved glycine at the V beta-J beta junction suggests the existence of diversity elements. The extracellular domains of the two axolotl beta chain constant region isotypes C beta 1 and C beta 2 show an impressively high degree of identity, thus suggesting that a very efficient mechanism of gene correction has been in operation to preserve this structure at least from the early tetrapod evolution. The transmembrane axolotl C beta domains have been less well conserved when compared to the mammalian C beta but they do maintain the lysine residue that is thought to be involved in the charged interaction between the TCR alpha beta heterodimer and the CD3 complex.

Ambystoma↗

Characterization of a multimeric polypeptide complex on the surface of thymus-derived cells in the Mexican axolotl.

We previously raised a rabbit antiserum (L12) against a 38 kD polypeptide which is expressed on the surface of thymocytes and peripheral T cells of an Urodele Amphibian, the Mexican axolotl (Ambystoma mexicanum). Here we show that L12 antibodies immunoprecipitate several labelled molecules from surface iodinated axolotl spleen cells, including the 38 kD molecule, but also two polypeptides of 43 and 22 kD which are covalently linked to other elements. Another rabbit antiserum (L10) was raised against detergent-solubilized axolotl thymocyte membranes and shown to recognize the majority of thymocytes and about half of the splenocytes in immunofluorescence. In Western blotting, L10 antibodies recognized a limited number of surface polypeptides in thymocyte and splenocyte lysates, including 43, 38, and 22 kD elements. Immune complexes formed between L10 antibodies and solubilized splenocyte membranes were used to immunize BALB/c mice intrasplenically in the aim of raising MoAbs specific for axolotl T cells. Monoclonal antibody 87.16 was shown to stain in immunofluorescence 26.7% of thymocytes and 26.8% of spleen cells. This MoAb recognized a 43 kD polypeptide that can covalently associate on the T-cell surface with several other molecules to form a multimeric complex.

Ambystoma mexicanum↗

T-cell-specific membrane antigens in the Mexican axolotl (urodele amphibian).

Comparative analysis of SDS-PAGE patterns of axolotl spleen cells membrane detergent lysates showed important discrepancies between control and thymectomized animals. Among these, a 38-kD protein band, which appeared as a major protein in controls, was not or poorly expressed after thymectomy. A rabbit antiserum (L12) raised against the 38-kD eluted band labeled in indirect immunofluorescence 80-86% of thymocytes and 40-46% of mIg- lymphoid cells in the spleen. The anti-38-kD antibodies stained in Western blotting two antigenically related polypeptides of 38- and 36-kD on splenocyte membrane lysates. Two-dimensional NEPHGE-PAGE analysis indicated that the anti-38-kD antibodies reacted in the spleen with several gathered spots in the 7.8-8.2 pI range, corresponding to 38-36-kD microheterogeneous polypeptides. Most of these spots are not further expressed in thymectomized animals. These results support evidence that the 38-kD surface antigens can be considered as specific surface markers of the axolotl thymus-derived lymphocytes.

Ambystoma mexicanum↗

High levels of HMG1-2 protein expression in the cytoplasm and nucleus of hydrocortisone sensitive amphibian thymocytes.

A major 26 kDa protein was identified in the cytoplasmic and nuclear compartments of axolotl thymocytes. A polyclonal antiserum was produced against the denatured form of this protein. High levels of 26 kDa were expressed by hydrocortisone-sensitive lymphocytes which represent a major thymocyte subpopulation in young animals. However, no further expression of the 26 kDa protein was observed in involuted thymus of adult animals nor in thymus of young artificially metamorphosed axolotls. The 26 kDa was never expressed by splenic and blood peripheral lymphocytes at any stage of development. Partial N-terminal amino acid sequence and amino acid composition demonstrate that the 26 kDa polypeptide is strongly homologous to HMG1-2 proteins, the most abundant members of the high mobility group (HMG) non-histone chromosomal proteins. HMG1-2 are thought to be involved in the organization of chromatin structure, as well as in the stability, replication and transcription of DNA. It was confirmed that the 26 kDa axolotl polypeptide is recognized by a well characterized rabbit antiserum to rat HMG1-2 proteins.

Ambystoma mexicanum↗

Surface markers of axolotl lymphocytes as defined by monoclonal antibodies.

In an attempt to identify urodele amphibian lymphocyte subpopulations by their surface markers, we prepared hybridomas from BALB/c mice spleen immunized with axolotl (Ambystoma mexicanum) blood and splenic leucocytes and purified immunoglobulins. Sixty-five hybridomas were selected and subsequently subcloned. Among numerous monoclonal antibodies (mAbs) thus obtained, four mAbs were extensively characterized by immunoblotting, single and double fluorescence and immunohistology. MAb 34.38.6 recognizes polypeptides between 65,000 and 72,000 MW and labels in immunofluorescence nearly all thymocytes, 60-63% splenic lymphocytes of normal animals but only 9% splenic lymphocytes in thymectomized animals. MAb 19.14.2 reacts with a 98,000 MW protein and labels a restricted lymphocyte population in thymus (52-77%) and spleen (20-25%). The immunohistological study demonstrates that 34.38.6 and 19.14.2 label most thymocytes and a large proportion of spleen leucocytes including lymphocytes, granulocytes and macrophages. In addition, 19.14.2 labels some large interdigitating cells in thymic epithelial areas and splenic cords. MAbs 33.45.1 and 33.101.2, respectively, recognize heavy (72,000-88,000 MW) and light (20,000-27,000 MW) axolotl immunoglobulin chains. They do not react with thymocytes but label a splenic lymphocyte population not labelled by mAb 34.38.6. The proportion of surface immunoglobulin-positive (sIg+) lymphocytes in spleen is not altered by thymectomy. MAb 33.101.2 labels 40-48% of splenic lymphocytes, 33.45.1 stains only 14% of these same cells. This suggests some interesting heavy-chain isotypic differences in axolotl. For the first time in urodele amphibians, mAbs differentiate T-like and B-like lymphocyte populations by their membrane markers. This will allow further analysis of the axolotl immune system.

Ambystoma mexicanum↗

[Spironolactone in idiopathic hirsutism, clinical and biological evaluation of treatment].

Ten midly hirsute women with normal serum levels of testosterone were treated with spironolactone (100 mg die), and the effectiveness of this dose-schedule regimen was evaluated after six months of therapy. Hirsutism subsided partly in all women. We estimated however, that in 6 of the 10 women, the clinical benefit was out weighed by the side effects and other drawbacks of a long term therapy and their medication was then stopped 6 months after its initiation. The changes induced on the pituitary function were evaluated by measuring basal serum T3 T4 and basal and stimulated (TRH-LHRH) TSH, PRL, FSH and LH, prior to and after one month of therapy. Spironolactone induced a greater response of TSH to TRH (p less than .025) but no change in T3 T4, PRL, FSH and LH. These data suggest that changes in TSH response are not explained by the speculated intrinsic oestrogenic activity of spironolactone but rather by a selective interaction with TSH secreting cells. We conclude that spironolactone therapy should not be recommended for the large category of normal women complaining of slight increase of facial hair.

Adult↗

The thyroid nodule: evaluation of fine-needle biopsy.

Sixty-three consecutive unselected patients with a solitary cold nodule of the thyroid were submitted to surgery. Prior to surgery they all had clinical evaluation and a fine-needle aspiration (FNA) biopsy of the nodule. Results of this study show that the FNA biopsy was correct in predicting cancer in 12 of 13 cancers for a sensitivity of 92%. When the nodule was benign, the FNA biopsy was right in 42 of the 50 benign nodules for a specificity of 84%. In comparison the clinical criteria alone were correct in suspecting only eight of the 13 cancers for a sensitivity of 62%, while correctly identifying 39 of the 50 benign nodules for a specificity of 72%. An association of the clinical criteria with the results of the FNA biopsy would have identified all the cancers in our group.

Adult↗

[Pattern and nomenclature of serum proteins of the amphibian urodele Ambystoma dumerilii Duges].

By means of many electrophoresis techniques, the serum of Ambystoma dumerilii was studied. The pattern in cellulose acetate is different from A. mexicanum proteinogram. Direct two-dimensional analysis in acetate/polyacrylamid gel with progressively increasing concentration from 4-30%, gives us the opportunity to observe 41 proteic components. By immunological results, the fastest proteic fraction is equivalent to a globuline with alpha-mobility more than an albumin. A nomenclature for this blood serum is proposed.

Ambystoma↗