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Y Sarret

Publications and source records attributed to Y Sarret.

10 recordsLinked to original sources
PubMed1992

Human keratinocyte locomotion: the effect of selected cytokines.

Epidermal growth factor (EGF) and basic fibroblast growth factor (bFGF) are two powerful mitogens for human keratinocytes that also have been shown to promote the healing of in vivo wounds. Transforming growth factor-beta (TGF-beta) markedly inhibits human keratinocyte proliferation and growth and yet has been shown to promote wound healing. Using a migration assay that evaluates pure cell locomotion independently from cell proliferation, we examined the influence of EGF, bFGF, and TGF-B on human keratinocyte locomotion. Although these agents had profound influences upon the growth potential of keratinocytes in parallel thymidine incorporation assays, they had no significant effect upon keratinocyte locomotion when cells were apposed to either tissue culture plastic or a collagen substratum. In contrast, we found that bovine pituitary extract (BPE), a poorly defined mitogen that is commonly used in keratinocyte cultures, could stimulate keratinocyte locomotion when the cells were apposed to a collagen substrate. These studies demonstrate that i) keratinocyte locomotion and proliferation operate by completely independent mechanisms, ii) the positive effects upon wound healing by EGF, bFGF, and TGF-beta are not due to a direct promotion of keratinocyte locomotion, and iii) that one or more components of BPE are capable of directly promoting keratinocyte locomotion on collagen.

Animals↗
PubMed1992

Constitutive synthesis of a 92-kDa keratinocyte-derived type IV collagenase is enhanced by type I collagen and decreased by type IV collagen matrices.

Human keratinocytes synthesize interstitial collagenase, a 72-kDa gelatinase, and a recently described 92-kDa gelatinase/type IV collagenase. We examined the synthesis of this novel enzyme by basal keratinocytes apposed to plastic, basement membrane collagen (type IV), and interstitial dermal collagen (type I). Samples of conditioned medium were electrophoresed on a 10% polyacrylamide, gelatin-ladened zymogram. Protein bands with gelatin-cleaving properties were identified by clarification of the gel and quantified by densitometry. A 92-kDa band had marked gelatinolytic activity and increased in culture over 72 h. The identification of this 92-kDa band as type IV collagenase was demonstrated by Western immunoblotting using monospecific antibody to the 92-kDa type IV collagenase. Keratinocytes apposed to type I collagen exhibited a threefold increase in the synthesis of the 92-kDa enzyme compared to cultures apposed to type IV collagen and a 1.5-times increase compared to plastic. The specificity of this enhancement was shown by constant levels of other proteins (e.g., the 72-kDa gelatinase). This study demonstrates that cell-matrix interactions modulate the synthesis of a recently described, keratinocyte-derived, 92-kDa gelatinase and that specific collagen types (I versus IV) have opposite effects upon the synthesis of this enzyme.

Collagen↗
PubMed1992

Keratinocyte migration is partially supported by the cell-binding domain of fibronectin and is RGDS-dependent.

Fibronectin (FN) plays a key role in cell attachment, embryonic development, and wound healing. In this respect, it is known that FN promotes keratinocyte migration. The aim of this study was to examine specific FN domains (120-kD cell-binding fragment, 45-kD collagen fragment, and 40-kD heparin fragment) and a biologically active peptide within the molecule (RGDS) for their ability to influence human keratinocyte (HK) locomotion. HKs were plated on gold salts coated with different substrates (type IV collagen, FN with or without the RGDS peptide, and the three FN fragments). After 20 h, locomotion tracks were quantified by computer-assisted image analysis that determines the area of each microscopic field occupied by migration tracks, a so-called migration index (MI). MIs on type IV collagen and FN were 39.14 +/- 2.8% and 30 +/- 0.4%, respectively. The maximal MIs on the collagen-binding domain and heparin-binding domain of FN were similar to our negative controls (plastic and albumin): 3 +/- 1%. In contrast, the maximal MI on the cell-binding fragment of FN was 18.45 +/- 2.1%. The effect of the cell-binding domain on keratinocyte motility was found to be dose dependent. Moreover, we could specifically inhibit the FN-driven locomotion using the RGDS sequence contained in the cell-binding fragment. We did not observe a synergistic effect (i.e., a higher MI) when we added the three fragments in a same dish. These results suggest i) that the cell-binding fragment of FN partially supports HK locomotion, ii) that other untested FN domain(s) should act in synergy with the cell-binding fragment to promote keratinocyte locomotion, or alternatively iii) that the FN function of promoting cell migration resides within the FN cell-binding domain, but the proper presentation of this domain to the cell requires an intact, native FN molecule, and iv) that the RGDS sequence is essential for HK movement.

Amino Acid Sequence↗
PubMed1992

[Fibronectins].

Fibronectins are complex glycoprotein macromolecules whose molecular weight is 440 kilodaltons. These proteins, found throughout the body, are soluble in biological fluids and insoluble in connective tissue. They contribute to tissue morphogenesis through multiple interactions with cells and extracellular matrix components. Fibronectins play a key role in cell attachment and remodeling during embryonic development, in cell migration and anchorage during wound healing, and in immune responses. This review discusses the structure of fibronectins, their integrin cell receptors, their biologic properties and their implications for diseases.

Cell Adhesion↗
PubMed1991

Salt-split human skin substrate for the immunofluorescent screening of serum from patients with cicatricial pemphigoid and a new method of immunoprecipitation with IgA antibodies.

In patients with cicatricial pemphigoid, immunoglobulins (usually IgG) and complement are deposited within the dermoepidermal junction and are detected by direct immunofluorescent staining of perilesional mucous membrane and/or skin with fluorescein-labeled antibodies to human immunoglobulins. Although rare, some patients also have circulating low-titer, anti-basement membrane zone autoantibodies. In this study, we report 11 patients with the clinical, histologic, and immunologic criteria for cicatricial pemphigoid who had circulating anti-BMZ autoantibodies as demonstrated by positive indirect staining of a normal human skin that had been fractured through the dermoepidermal junction by prolonged incubation in a cold, 1 mol/L sodium chloride solution. On this salt-split human skin substrate, 9 of the 11 patients (82%) had autoantibodies that bound to the epidermal roof, one serum stained only the dermal floor, and one serum stained both sides of the separation. The predominant class of immunoglobulin in the patients' sera that bound to the substrate was IgA; IgA was the single immunoglobulin in 55% and was associated with IgG in 18%. IgG was the only immunoglobulin detected in 27% of the cases. No specific protein was detected by either Western immunoblot or a new IgA immunoprecipitation procedure.

Adult↗
PubMed1991

Autoimmunity to type VII collagen.

The epidermolysis bullosa acquisita (EBA) antigen is type VII collagen that is found within the anchoring fibrils of the basement membrane zone beneath stratified epithelia. Autoimmunity to the EBA antigen/type VII collagen has been associated with three diseases: EBA, bullous systemic lupus erythematosus (SLE) and a subset of linear IgA bullous diseases. Although some systemic diseases in which autoimmunity is thought to play a role (eg, inflammatory bowel disease, thyroiditis) have been reported in association with EBA, so far there have not been systemic diseases associated with autoimmunity to type VII collagen other than SLE. In the case of EBA and bullous SLE, it appears that many patients may have a genetic predisposition toward autoimmunity because they share a human leukocyte antigen (HLA) major histocompatibility (MHC) class II cell surface protein, HLA-DR2. The full clinical spectrum of EBA and perhaps other diseases in which there is an association with autoimmunity to type VII collagen is currently being defined.

Animals↗
PubMed1990

GP37 is different from filaggrin.

The identity of two differentiation markers of human epidermis, filaggrin and a Concanavalin A (Con-A) reactive glycoprotein of 37 kD, has been studied. Human epidermis was extracted in Nonidet P-40 buffer, and the soluble proteins were separated by two-dimensional electrophoresis. Con-A reactive glycoproteins were identified by incubating gels with iodinated lectin followed by autoradiography. Identical, parallel gels were electrophoretically transferred to nitrocellulose paper and filaggrin-related molecules labeled by the specific monoclonal antibody AKH1. We found that the 37-kD Con-A reactive component was resolved by two-dimensional gel electrophoresis into several glycoproteins and that the lectin Con-A does not bind to filaggrin. Under these conditions, the anti-GP37 serum failed to identify any component. However, when applied to human keratinocyte culture extract, AKH1 and the anti-GP37 serum reacted in a similar way. These data show 1) that the 37-kD band is not homogeneous but contains distinct markers of differentiation (filaggrin and Con-A reactive glycoproteins) and 2) that the GP37 antibody's specificity is for the filaggrin precursor.

Antibodies, Monoclonal↗
PubMed1989

Bullous pemphigoid and cicatricial pemphigoid: immunoblotting detection of involved autoantigens.

Bullous pemphigoid (BP) and cicatricial pemphigoid (CP) are subepidermal bullous autoimmune diseases which have distinct clinical features but identical immunological status. In order to determine whether these diseases could be dissociated on the basis of qualitative differences in serum antibodies to basement membrane zone (BMZ) antigens, the reactivity of sera from 7 CP and 29 BP patients with proteins extracted from normal human epidermal sheets (containing most of the lamina lucida components) was analysed using immunoblotting and compared to that of 10 normal sera. 20 out of the 29 BP sera contained antibodies recognizing one or several protein(s) of 240, 200, 180 and 165 kD molecular weight (MW). Antibodies in 4 out 7 CP sera specifically reacted with one or two polypeptides of 240 and 120 kD MW. These data confirm the heterogeneity of BP antigens and show the presence in CP of a novel 120 kD MW polypeptide which is found only in CP but not in BP. Taken together these findings demonstrate that in BP and CP, autoantibodies are directed to both common and specific BMZ antigens, their physiopathological significance need to be understood.

Adult↗