PubMed Health⌕ Search

Biomedical subjects

D Herbage

Publications and source records attributed to D Herbage.

At least 55 records · Page 3Linked to original sources
PubMed1990

T cell regulation of collagen-induced arthritis in mice. I. Isolation of Type II collagen-reactive T cell hybridomas with specific cytotoxic function.

After immunization with native type II collagen (CII), susceptible strains of mice (H-2q) develop a polyarthritis that mimics rheumatoid arthritis. Although the underlying mechanisms are still undefined, T cells and particularly CD4+ lymphocytes seem to play a crucial role in the initiation of collagen-induced arthritis. To investigate whether CD8+ cells may participate in the pathogenesis of the disease, we have generated lines and clones of cytotoxic T cell hybridomas reactive to CII by fusion of lymph node and spleen cells from bovine native CII-primed C3H.Q (H-2q) mice and the AKR-derived thymoma cell line BW 5147. Clones were selected for their ability to lyse syngeneic macrophages pulsed with bovine native CII in an Ag-dependent manner. The two hybrid clones that were characterized, exhibited cell surface phenotypes of cytotoxic cells and reacted with CII purified from various species. However, each of them recognized different determinants on the CII molecule. P3G8 clone was specific for an epitope shared by CII and type XI collagen, whereas P2D9 clone reacted with CII and type IX collagen. Both hybridomas recognized CII-pulsed targets in association with H-2Kq molecules. These data indicate that the two CII-specific cytotoxic clones recognize different epitopes that are shared by other articular collagens and will allow us to test their influence on the development of arthritis in vivo.

Animals↗
PubMed1990

Comparative analysis of collagens solubilized from human foetal, and normal and osteoarthritic adult articular cartilage, with emphasis on type VI collagen.

The different collagen types were extracted sequentially, by 4 M guanidinium chloride and pepsin, from human foetal and normal and osteoarthritic adult articular cartilage. They were characterized by electrophoresis and immunoblotting. Most of the collagenous proteins present in articular cartilage from young human foetuses were solubilized: almost 40% of the total collagen was extracted in the native form with 4 M guanidinium chloride. Type VI collagen was detected in this fraction as high-molecular-mass chains (185-220 kDa) and a low-molecular-mass chain (140 kDa). Type II, IX and XI collagens were also present, but were extracted more extensively by pepsin digestion. Comparative analysis of normal and osteoarthritic cartilage from adults reveals some major differences: an increase in the solubility of the collagen and modifications of soluble collagen types in osteoarthritic cartilage. Furthermore, type VI collagen was present at a higher concentration in guanidinium chloride extracts of osteoarthritic cartilage than those of normal tissue. This finding was corroborated by electron microscopic observations of the same samples: abundant (100 nm) periodic fibrils were observed in the disorganized pericellular capsule of cloned cells in osteoarthritic cartilage. In normal tissues the pericellular zone was more compact and contained only a few such banded fibrils. The differences in the collagen types solubilized from normal and osteoarthritic cartilage, although corresponding to a minor proportion of the total collagen, demonstrate that important modifications in chondrocyte metabolism and in the collagenous network do occur in degenerated cartilage.

Aged↗
PubMed1990

Arthritogenicity of minor cartilage collagens (types IX and XI) in mice.

Native type II collagen, the major cartilage collagen, is immunogenic and arthritogenic in rodents. To investigate whether minor cartilage collagens are arthritogenic, we immunized DBA/1 mice with the pepsin-soluble fractions of type IX or type XI collagen emulsified in Freund's complete adjuvant. Both collagens were arthritogenic in DBA/1 mice after only 1 injection. However, the incidence of the polyarthritis was lower and the severity was lesser than with that induced by bovine type II collagen, even when a booster injection was administered. All mice developed a humoral response to the immunizing antigen, without any relationship to the arthritic status. Interestingly, competition experiments showed that antibodies raised against type XI collagen also bound with high avidity to type II collagen. In contrast, sera from type IX collagen-immunized mice did not react with either type II or type XI collagen. We conclude that types IX and XI minor cartilage collagens are both arthritogenic and immunogenic in DBA/1 mice. Whether the recognition of epitopes common to different collagens is relevant to the articular pathology remains to be elucidated.

Animals↗
PubMed1990

Use of the acyl azide method for cross-linking collagen-rich tissues such as pericardium.

Collagen biomaterials should be cross-linked in order to prevent biodegradation when they are used as implants. We have compared the cross-linking efficiencies of glutaraldehyde and acyl azide in pericardium. Glutaraldehyde is used currently, but it elicits a cytotoxic effect which reduces the biocompatibility of cross-linked tissue. We have attempted to overcome this problem by developing a cross-linking method that obviates incorporation of foreign agents. Our process involves transformation of free carboxyl groups on collagen into acyl azide groups, which react with free amino groups on adjacent side chains. We have shown that the greatest increase in the thermal stability of collagen, as measured by differential scanning calorimetry, is achieved when tissue swelling is inhibited by the addition of sodium chloride (1 M) during acyl azide formation. Under these conditions, the denaturation temperature (Td) of pericardial collagen treated with acyl azide is raised to 83.4 degrees C and that of tissue treated with glutaraldehyde to 85.1 degrees C. Moreover, acyl-azide-treated tissues have the same resistance as glutaraldehyde-treated tissues to chemical solubilization by cyanogen bromide and to enzymatic digestion by collagenase.

Acylation↗
PubMed1990

Tissue reaction to subcutaneous implantation of a collagen sponge. A histological, ultrastructural, and immunological study.

The biocompatibility of a subcutaneously implanted bovine collagen sponge (Haemostagen) was studied in rats by analyzing tissue reactions up to 3 months by histological and ultrastructural methods; in addition, the level of serum antibodies to bovine type I collagen (the major implant collagen) was measured by solid-phase radioimmunoassay. By 8 h after implantation, the implant was completely filled with polymorphonuclear cells (PMNs). After 8 days, fibroblasts had developed a granulation tissue within the sponge and the PMNs had almost disappeared. The small residue that remained after 1 month consisted of some densely packed collagen fibrils containing giant cells, which had disappeared by 3 months. No antibodies to bovine type I collagen were found in the sera of implanted rats.

Animals↗
PubMed1989

Immunohistochemical study of the biological fate of a subcutaneous bovine collagen implant in rat.

The biological fate of a bovine collagen implant (Zyderm Collagen Implant ZCI), injected subcutaneously into rats, was studied by the immunoperoxidase technique using specific antibodies against the bovine implant and against types I, III, IV, V collagens, fibronectin and elastin. The implant remained in the animals until the end of the experiment (90 days), with no visible modification, as demonstrated by immunoperoxidase labelling and scanning electron microscopy. A slight inflammatory reaction was visible around the implant 24 h after injection and within the implant 3 days after injection. Fibroblast invasion began 7 days after injection. The chronology of the deposition in the implant of the host (rat) extracellular matrix components was as follows: by 24 h after injection, fibronectin was observed throughout the implant; types I and V collagens appeared on the 7th day, and, in contrast to surrounding connective tissue, type V collagen labelling was obtained without acid pretreatment of the section. Types III and IV collagens were detected inside the implant only 30 days after injection. At the end of the experiment (90 days), there was abundant types I and IV collagens after fibroblast migration, and abundant type IV collagen demonstrating an important vascularization. No elastic fibres could be detected inside the implant but they appeared as a dense network around the implant in host connective tissue.

Animals↗
PubMed1989

Effects of tiaprofenic acid on interleukin 1, phospholipase A2 activity, prostaglandins, neutral protease, and collagenase activity in rheumatoid synovial fluid.

IL-1 and prostaglandin (PGE2, PGF2 alpha, TXB2) concentrations, PLA2 activity, neutral protease activity, and collagenase activity specific for types I and II collagen were determined in the SF of patients suffering from RA, before and after treatment with TA. Active and latent forms of protease and collagenases were regularly detected but were unrelated to IL-1, PLA2, and PGE2. TA induced a significant decrease in tested eicosanoids but IL-1, PLA2, and proteases were unchanged.

Adult↗
PubMed1989

Problems in the immunolocalization of type IX collagen in fetal calf cartilage using a monoclonal antibody.

Monoclonal antibodies were prepared against the pepsin-resistant fragments (X1-X3) of bovine type IX collagen. One of the five hybridomas that gave a positive reaction in an enzyme-linked immunosorbent assay was selected (H1a) for structural analysis and immunolocalization of type IX collagen. The location of the epitope for H1a was deducted from immunoblots and electron microscopic observations after rotary shadowing. The H1a antibody binds to one end of the longest X2, X3, X4 molecules, and preferentially 40-55nm from one end of X1 molecules thus, on or near the noncollagenous domain, NC2. Different immunolocalizations of type IX collagen in the superficial, middle and deep zones of fetal calf epiphyseal cartilage were observed depending on the thickness of the section and on hyaluronidase digestion conditions. In the middle and deep zones, staining with H1a throughout the matrix was obtained only with thin sections (5 microns) and digestion for 1 h at 37 degrees C. With thick sections (15 microns) or with digestion for 1 h at 24 degrees C, staining was restricted to the pericellular regions. Staining throughout the matrix was obtained in the superficial zone under all experimental conditions. Without hyaluronidase treatment, no immunofluorescent staining was seen with either H1a or polyclonal antibody to type II collagen, indicating that type IX collagen is present throughout the matrix in the different zones of fetal calf cartilage. This result is in good accordance with the recent demonstration of common cross-links between type II and type IX collagen in chicken and bovine cartilage. However, the preferential unmasking of type IX collagen antigenic sites in the pericellular regions of middle and deep zones of fetal calf cartilage does not preclude the presence in that region of a special pericellular organization of the collagenous network.

Animals↗
PubMed1988

Antibodies to types I, II, IX, and XI collagen in the serum of patients with rheumatic diseases.

Antibodies to native types I, II, IX, and XI collagen were measured, using a 125I-solid-phase radioimmunoassay, in serum from 104 patients with rheumatic diseases (rheumatoid arthritis, osteoporosis, Paget's disease, or osteoarthritis). In all disease groups, antibodies to type II collagen occurred with greater frequency than antibodies to type I collagen (11-35% versus 5-23%). Antibodies to type XI collagen were the most frequent: They were present in approximately 50% of the patients in the rheumatoid arthritis, Paget's disease, and osteoporosis groups. Antibodies to type IX collagen were found at a high frequency in the rheumatoid arthritis group only (44%). Analysis of the clinical data suggested that the presence of antibodies to collagen was associated with disease that was less severe or of shorter duration.

Adult↗
PubMed1988

Distribution and organization of the elastic system fibres in healthy human gingiva. Ultrastructural and immunohistochemical study.

The ultrastructural distribution and organization of the elastic system fibres, i.e. oxytalan, elaunin and elastic fibres, were studied by transmission electron microscopy and by an immunohistochemical method for the detection of elastin in healthy human gingiva. The morphological distribution of these fibres was characterized by the presence of oxytalan, elaunin and elastic fibres, respectively, in the upper, medium, and deep layers of gingival connective tissue. Anti-elastin antibody reacted with microfibrils and amorphous material of the elastic system fibres throughout the gingival connective tissue. These findings were interpreted as indicating that the microfibrils were associated with small amounts of elastin at their surface.

Adult↗
PubMed1988

Effect of praziquantel on hepatic fibrosis in experimental Schistosomiasis mansoni.

The effect of specific chemotherapy on hepatic fibrosis was studied in Swiss albino mice infected with Schistosoma mansoni. The effect of treatment with praziquantel at 8, 12, and 20 weeks postinfection on fibrosis was assessed by determination of total hepatic collagen content, ratio of type III/I + III collagen, granuloma volume, and histopathological examination of liver section. Total collagen content was reduced in mice treated at the 8th week of infection compared to respective infected controls. The ratio of type III/I collagen was within normal limits. The decrease in collagen deposition was not observed when treatment was given 12 or 20 weeks postinfection. Early specific treatment of schistosomiasis may be important in the therapeutic approach to the control of morbidity in schistosomiasis.

Animals↗
PubMed1987

Comparative structural studies of reconstituted and native type I and type II collagen fibrils by low-angle X-ray diffraction.

Acid-soluble and pepsin-soluble type I collagen from calf skin and pepsin-soluble type II collagen from bovine articular cartilage were precipitated in fibrillar form by various methods. Reconstituted native-like fibrils were analysed by low-angle X-ray diffraction, and the patterns were compared with those obtained with native type I (rat tail tendon) and type II fibrils (bovine articular cartilage). For both orientated and disorientated forms of these samples, we measured the ratio of the first/third-order intensities of the meridional diffraction peaks which are associated with the gap-filling. The values obtained with the reconstituted native-like fibrils (types I and II) were double and 20-times the values, respectively, measured for rat tail tendon and bovine articular cartilage. These differences reflect the extent of specific interactions of other components (proteoglycans) at the gap level along the collagen fibrils in the two tissues.

Animals↗
PubMed1987

Wound healing of human skin transplanted onto the nude mouse. II. An immunohistological and ultrastructural study of the epidermal basement membrane zone reconstruction and connective tissue reorganization.

The reconstruction of human epidermis during healing of human skin wounded after grafting onto the nude mouse was described in a previous paper (M. Démarchez, P. Sengel, and M. Pruniéras, 1986, Dev. Biol. 113, 90-96). The regeneration of the epidermal basement membrane zone (BMZ) and the reorganization of the connective tissue are the subjects of the present study. They were investigated by two complementary methods: electron microscopy to analyze the BMZ reorganization, and indirect immunofluorescence with species-specific and cross-reacting antibodies directed against laminin, bullous pemphigoid antigen, mouse or human collagens of types I or IV, human elastic fibers, fibronectin, fibrin, actin, and human vimentin, to examine the species origin and distribution of BMZ and connective tissue components during the regeneration process. It is reported that grafted human skin preserves its own immunological markers not only in the epidermis but also in the BMZ and dermis as well, and that, after injury, its regeneration proceeds according to the following sequence of overlapping events: production of a mouse granulation tissue; reepidermization by human cells; reconstruction of a BMZ with human characteristics; formation of a human neodermis. It is concluded that human skin grafted onto the nude mouse is able to regenerate its three structural compartments, namely, the epidermis, BMZ, and dermis. Interestingly, it appeared, also, that the connective tissue regeneration would be a two-step mechanism including the sequential formation of two tissues of distinct sources, namely, a granulation tissue and a neodermis.

Animals↗
PubMed1987

Ultrastructural immunolocalization of elastic fibers in rat blood vessels using the protein A-gold technique.

Identification of elastic fibers at the ultrastructural level is accomplished by a post-embedding immunohistochemical technique using the protein A-colloidal gold method. Antisera against elastins from human dermis and rat aorta have been characterized by radioimmunoassay and then applied to thin sections of rat blood vessels. Two fixative solutions and two embedding media have been tested. Both antibodies bind to elastic fibers of normal arteries and veins, indicating crossreactions among organs and species. The high sensitivity of this method is demonstrated by its application to the detection of neo-elastogenesis in the intimal thickening of aortic grafts.

Animals↗
PubMed1986

Type IX collagen is a potent inducer of PGE2 and interleukin 1 production by human monocyte macrophages.

Type IX collagen and its collagenous fragments are potent stimulatory agents on human blood mononuclear cells for the production of prostaglandin E2 and interleukin 1/mononuclear cell factor. Type IX collagen is 2-4-fold more potent that type I and II and 1 alpha-, 2 alpha- and 3 alpha-collagens. This property may be important in the destructive process of cartilage in inflammatory diseases.

Cells, Cultured↗
PubMed1986

Effect of malotilate on chronic liver injury induced by carbon tetrachloride in the rat.

The effect of malotilate, a new drug proposed for the treatment of chronic liver diseases, was studied in carbon tetrachloride (CCl4)-induced chronic liver injury in the rat. Treatment with CCl4 (0.5 ml/kg twice per week, intraperitoneally for 6 or 9 weeks) led to marked necrosis, steatosis and fibrosis, as shown by both biochemical and histological examinations, and a significant decrease of the bromosulfophtaleine (BSP) clearance test. Malotilate (50 mg/kg p.o., 5 days per week given simultaneously with CCl4 for 6 weeks), suppressed the increase of plasma aminotransferase activity and decreased significantly the accumulation of lipid and collagen in the liver; histology confirmed this protective effect of malotilate. The BSP clearance test returned to normal values and the rise in hepatic collagen synthesis activity in the malotilate-treated and intoxicated rats was reduced as compared with intoxicated control rats. The same effect was found when malotilate (100 mg/kg, p.o., 5 days per week), was given for 3 weeks to rats already intoxicated during the 6 previous weeks. Malotilate was able to prevent the increase of hepatic alterations that appeared during the last 3 weeks of CCl4 intoxication. These results show clearly that malotilate can markedly reduce the hepatic disorders induced by a chronic CCl4 intoxication in the rat.

Animals↗
PubMed1986

Collagen expression during teratocarcinoma cell line-induced endochondral bone tumor.

Collagen immunotyping by indirect immunofluorescence was performed in order to investigate the sequential development of bone formation. Osseous tumors were obtained after subcutaneous injection of 3/A/1D-1 teratocarcinoma cell line into 129/Sv mice (Nicolas et al., 1980). Frozen sections of developing tumors were incubated with specific antibodies directed against Types I, II, III, IV, and IX collagens. On Day 9, the expression of Type I and Type III collagens was correlated with the proliferation of mesenchymal cells. From Day 10, chondrogenesis was characterized by the occurrence of cartilaginous collagens, Types II and IX, in the cartilage matrix. Type IV collagen was also detected in focal areas and revealed vascular invasion of the tumor. On Day 13, osteogenesis was demonstrated by the presence of Type I collagen in the bone matrix coating the surfaces. Immunolocalization of Type III collagen on the hemopoietic elements corresponded with the bone remodeling. The sequential transitions of collagen types confirm the development of an endochondral bone tumor. These results suggest that 3/A/1D-1 teratocarcinoma cell line constitutes a valuable system for in vitro study of endochondral bone formation and cell differentiation.

Animals↗