[Serum assay of the P1 fragment of human laminin].
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Biomedical subjects
Publications and source records attributed to D J Hartmann.
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Serum immunoglobulin levels IgG, IgA, IgM were assayed by laser-nephelometry and IgD by a liquid phase radio-immunoassay in 133 patients with monoclonal gammopathy. Our result show that IgD concentration decrease in parallel with polyclonal immunoglobulin levels. We observe a decrease of IgD level, more pronounced in malignant gammopathy in which the polyclonal immunoglobulin levels are usually low.
Odontoblast-like cells derived from human tooth pulps were maintained in explant culture and grown either on glass coverslips only (used as control) or on glass coverslips coated with cyanoacrylate films. Ultrastructural and cyto-morphometric evidence showed that cells exposed to cyanoacrylate, in contrast to controls, display a significant decrease of rough endoplasmic reticulum and mitochondria. In addition, immunofluorescent staining and radioimmunoassays for type-I collagen suggested disturbances in production for the exposed cells. The use of anti-fibronectin antibodies with electron-microscopic immunoperoxidase-labelling demonstrated that the adherence of cells to cyanoacrylate can involve both adhesion plaques and fibronectin. These results therefore suggest that there were no apparent differences in the adhesion interaction of cells between glass and cyanoacrylate substrates.
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.
We describe a sensitive liquid phase radioimmunoassay for serum IgD. Extreme values obtained from 85 control patients sera are 0.2 and 121 mg/l with an arithmetic mean of 25 mg/l. In atopic patients (with high serum IgE levels), arithmetic mean is 47 mg/l.
Section of the anterior cruciate ligament of the right knee was performed in 20 rabbits which were sacrificed 3 months later. During that time, 10 animals were treated with a proteino-polysaccharide compound. The importance of cartilage destruction was evaluated by measuring the articular surfaces and the surface of the fissured areas using an automatic image analyzer; osteophytes were measured by a volumetric study. The percentage of the fissured areas appears to be without correlation with the osteophytosis, the increase of the water content or the decrease of glycosaminoglycans following the procedure. In animals which were treated, the extent of the fissure is significantly reduced but the biochemical and histometric modifications are not too different from those of the reference group.
The authors have developed a solid phase radioimmunoassay for the detection of antibodies to collagens. They give the technical data of this assay and some results mainly in rheumatic diseases.
We analysed epithelial-mesenchymal interactions that occur during the early stages of the formation of mouse molar roots using light and electron microscopy. Morphological changes observed in the cells of Hertwig's epithelial sheath, the pulp and the follicular mesenchyme are described. The Hertwig's epithelial cells lose their cuboidal form and become flattened, apparently intermixing with the cells of the follicular mesenchyme. At the light- and electron-microscope levels, immunoperoxidase techniques were used to localize fibronectin, laminin and type-IV collagen. These appear to be closely associated with cell differentiation and matrix deposition in developing tooth roots. In addition, at the ultrastructural level, intracellular immunoreactivity was detected. The rough endoplasmic reticulum and nuclear envelope of some cells of the periodontal ligament facing the acellular cementum exhibited specific reactivity with laminin and type-IV collagen. Moreover, these periodontal ligament cells express keratin, but not vimentin, filaments. Our results demonstrate that Hertwig's epithelial cells maintain their capacity to synthesize laminin and type-IV collagen, as well as to express keratin filaments, despite basement membrane fragmentation and the disorganization of Hertwig's epithelial sheath. Thus, some Hertwig's epithelial cells remain in the periodontal ligament intermixed with follicular mesenchyme cells.
We studied the levels of carcinoembryonic antigen (CEA) and carbohydrate antigen 19-9 (CA 19-9) in 90 patients with primary lung cancer. One or both markers were increased in 45 patients (50%): CEA only was increased in 13, CA 19-9 only was increased in 19, and both CEA and CA 19-9 were increased in 13. Increase of markers did not differ according to histologic subtype of cancer. Increase or decrease of the markers (mainly CA 19-9) usually parallelled evolution of the disease in our patients. Thus, measurement of both CEA and CA 19-9 levels are of diagnostic value in half the patients with primary lung cancer.
Fetal calf serum and a serum substitute, Ultroser G, were compared for their effects on the growth curves, clonal growth and cell cycle progression of rabbit chondrocytes in primary culture and during at least three cell passages and included a screen for the maintenance of cartilage-like differentiation i.e. the presence of type II collagen. Proliferation was also compared with another serum substitute, Nu-Serum. Ultroser G is shown to be equivalent to fetal calf serum as far as chondrocyte proliferation is concerned, clonal growth is improved and biosynthesis of type II collagen is maintained in primary culture.
A solid-phase radioimmunoassay using 125I-protein A is described for the detection of antibodies to collagens of different types. The optimal conditions for the adsorption of collagen onto polystyrene microplates, then the incubations with the antiserum and finally with the 125I-protein A have been evaluated. The technique was applied successfully to antisera raised in rabbit, goat, guinea pig and mouse against human type I, II, III, IV, V and bovine type I, II, 1 alpha 2 alpha 3 alpha, X1-X7 collagens.
The present paper describes the optimisation of a liquid phase [125I] C1q binding assay for the detection of circulating immune complexes. Blood must be collected without any anticoagulant. The serum can be stored or transported as usual without damage. A 90 min incubation is needed to reach the equilibrium of the reaction; polyethylene glycol (2,5% final concentration) is added to precipitate the [125I] C1q-CIC complex. By its simplicity and reproducibility, this test can be currently employed, especially in the case of treatment by plasmapheresis.
We report a radio-immunoprecipitation method for the measurement of circulating insulin antibodies using porcine, bovine or human radiolabeled insulin and precipitation of the bound fraction by a combined polyethylene glycol--second antibody method. The coefficient of variation for intra- and inter-assay is below 8,5 p. cent and 17 p. cent respectively. 100 insulin treated patients sera were tested in the three systems: the results showed a great scattering; for the same serum, the values obtained with porcine and human insulins are below that with bovine insulin. The data correlated statistically significantly with those found with a commercial kit (r = 0,97, n = 170, t = -4,91, p greater than 0,001).
Fibronectin (by laser nephelometry) and collagen IV blood levels (by radioimmunoassay) were studied in 183 diabetics and compared with 101 non diabetic patients. Diabetics have more collagen IV and less fibronectin than non diabetics. Divergence has increased since 20 years old; fibronectin blood levels is always low in diabetics, even in the young. Collagen IV is higher in diabetics with angiopathy, and specially if insulin dependent. Diabetics which have a good control have normals levels. In the other hand when HBA1C greater than or equal to 9%, collagen IV blood level increases quickly and fibronectin level decreases. The importance of the antidiabetic treatment is underlined.
New minor disulfide-bonded collagens were recently described in cartilaginous tissues. The localization of these molecules in epiphyseal proper and growth plate cartilage of fetal calf has been studied using light and electron immunoperoxidase microscopy. The labeling was restricted to the pericellular region of the chondrocytes with an increasing intensity form the superficial to the inner zone of the epiphysis. At the ultrastructural level, the fine-non striated fibrils of the pericellular matrix were stained, demonstrating their collagenous nature. These minor collagenous chains are thus new components of the chondrocyte "exoskeleton" in which other molecules (proteoglycans, chondronectin, fibronectin and type V collagen) have been previously demonstrated.
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In the dermis of inter-appendage and glabrous skin, interstitial collagen types I, III, and V are abundant, while fibronectin is scarce. Conversely, in the morphogenetically active foci of cutaneous appendages, interstitial collagen is scarce or absent, whereas fibronectin is abundant. Type IV collagen and laminin are localized at the dermal-epidermal junction and distributed evenly.
Immunological assays appear to be the only alternative to the microbiological method for analysis of pantothenic acid in foods and blood. In order to evaluate the influence of the linker on the immunogenicity of the hapten, we have tried to raise antisera against pantothenic acid in rabbits using different conjugates. The hapten was coupled to a carrier protein (BSA or thyroglobulin) using adipoyl dichloride (adipoyl conjugate) or bromoacetyl bromide (acetyl conjugate). Only the acetyl conjugate has induced the production of a specific antibody. With this antibody, an assay on microplate using the ELISA inhibition technique was developed to measure pantothenic acid. The use of pantothenic acid coupled to thyroglobulin with adipoyl dichloride as the capture antigen has improved the sensitivity of the ELISA. This assay was applied to food products and blood.