Fibrillin Immunoreactive Fibers Constitute a Unique Network in the Human Dermis: Immunohistochemical Comparison of the Distributions of Fibrillin, Vitronectin, Amyloid P Component, and Orcein Stainable Structures in Normal Skin and Elastosis

Fibrillin, a 350-kD glycoprotein, was recently localized to elastin-associated 10nm microfibrils. Here, the distribution of fibrillin immunoreactivity was determined in normal skin in individuals of different ages and in lesions of solar elastosis or anetoderma. It was compared with the distribution of orcein-stainable fibers and with the immunoreactivities of vitronectin and amyloid P component. These glycoproteins are known to occur in conjunction with the orcein-stainable elastic fibers in adults, but not in the young. Fibrillin immunoreactivity was associated with orcein-stainable fibers in normal skin of both adults and the young. In addition, the fibrillin immunoreactive fiber network comprised fine fibers that were unstainable by orcein, anti-vitronectin, or anti- amyloid P component. Such fine fibers were especially abundant close to the dermal-epidermal junction zone. Immunoreactivities of anti-vitronectin and anti-amyloid P component were not always associated with fibrillin immunoreactivity but were consistently found to co-localize with orcein-stainable fibers in adults. This suggests vitronectin and amyloid P component to be associated with the amorphous elastin rather than with the microfibrils, although alternative interpretations are possible. In elastotic lesions, fibrillin immunoreactivity was generally fainter than that obtained using anti-vitronectin or anti-amyloid P component. In contrast, an extensive network of dermal fibers stained by anti-fibrillin, but not by anti-amyloid P component, anti-vitronectin, or orcein, was seen in an anetoderma lesion. In conclusion, fibrillin immunoreactivity is associated with a unique dermal network, which ultrastructurally is composed of microfibrils. These fibers are proposed to have an important structural and functional role in anchoring the dermal elastic fibers in the extracellular matrix and to the lamina densa

Human γ-trace. Primary structure, localization to neuroendocrine cells and concentration in body fluids

γ-Trace is a low molecular weight human protein with unknown function, occurring in body fluids. this thesis deals with the determination of the complete amino acid sequenceof human -trace, its localization to neuroendocrine cells and concentration in human body fluids at various ages.γ-Trace was isolated from human urine, and antisera specific for -trace were raisedin rabbits.An enzyme immunoassay, sensitive down to...

Production, characterization and use of monoclonal antibodies against the major extracellular human cysteine proteinase inhibitors cystatin C and kininogen

Murine monoclonal antibodies against the major cysteine proteinase inhibitors of human biological fluids, cystatin C and kininogen, were produced. The cystatin C antibody, HCC3, with a Ka of 2times107 l/mol, increased the inhibition of papain by cystatin C and was suitable for use in immunoblotting, immunohistochemistry and in the construction of a sensitive sandwich enzyme immunoassay for quantification of cystatin C. It recognized not only free cystatin C but also cystatin C in complexes with cysteine proteinases. The kininogen antibody, HK4, was directed against the third, cysteine proteinase inhibitory domain of the heavy chain of kininogen (Ka=1times107 l/mol), but did not influence the papain inhibitory activity of kininogen. It reacted with free kininogen as well as kininogen in complex with cysteine proteinases. Both antibodies could be used for the production of specific immunosorbents

Renal arteriovenous shunting in rejecting allograft, hydronephrosis, or haemorrhagic hypotension in the rat

We studied the occurrence of arteriovenous (A-V) shunting in three experimental rat models, namely in rejecting allograft kidney, in uni- or bilateral ureteral obstruction, and in haemorrhagic hypotension. Isografted or sham-operated rats served as controls. Radiolabelled microspheres were injected into the renal artery and the increase in the amount of radioactivity in the lungs was considered to reflect A-V shunting in the kidney. In animals exposed to haemorrhage, with a blood pressure not less than 70% of the initial blood pressure, practically no shunting was seen. When animals were bled to a hypotension beyond the autoregulation, A-V shunting occurred inversely correlated to the degree of hypotension. In ureteral obstruction, a less marked but significant increase in shunting of microspheres to the lungs was found after 24 h of unilateral obstruction, irrespective of whether the spheres were injected into the obstructed or the contralateral kidney. Significant A-V shunting during the allograft rejection process was also demonstrated. Histologically, microspheres were found in afferent arterioles less frequently in kidneys with A-V shunting than in controls. These results indicate that A-V shunting is involved in haemorrhagic hypotension, renal graft rejection, and hydronephrosis. In the latter situation A-V shunting is probably regulated by a humoral factor. © 1994 European Dialysis and Transplant Association-European Renal Association

Abnormal Metabolism of γ-Trace Alkaline Microprotein : The Basic Defect in Hereditary Cerebral Hemorrhage with Amyloidosis

ALTHOUGH the total incidence of cerebral hemorrhage is high, comparatively few reports concerning the familial occurrence of this disease have been published.1,2 In 1935 Arnason described 10 families with a high incidence of cerebral hemorrhage and concluded that a hereditary form of the disease was present in these families.3 Further clinicopathological investigations of the disease revealed an autosomal dominant inheritance and a connection between the disease and a special form of amyloidosis confined to the cerebral vasculature.4 This type of cerebral hemorrhage is therefore generally referred to as hereditary cerebral hemorrhage with amyloidosis. Recently, the fibrillar components of the amyloid

Demonstration and classification of amyloidosis in needle biopsies of the kidneys, with special reference to amyloidosis of the AA-type

To examine whether sequence-specific antibodies directed against serum amyloid A were useful in the demonstration and classification of amyloidosis, needle biopsy specimens from the kidneys of 152 cases with renal disorders were investigated using the avidin-biotin-peroxidase complex technique of immunohistochemistry. A distinct immunoreactivity of protein AA was seen in biopsies from all 42 individuals who were clinically classified as having the AA-type of amyloidosis. The stained areas coincided with deposits stained by Congo red. Four of these cases demonstrated immunoreactivity of both protein AA and light immunoglobulin chains and all biopsies except one showed immunoreactivity for the amyloid P-component. After treatment with potassium permanganate, the amyloid deposits in the biopsies of all 42 cases lost their affinity for Congo red. Ten patients with clinical and laboratory findings compatible with the AL-type of amyloidosis were also investigated. All their biopsies demonstrated Congophilic amyloid deposits but none of them showed any immunoreactivity of protein AA. Amyloid deposits of lambda light immunoglobulin chains-but not kappa-were demonstrated in biopsies from four patients. The amyloid P-component was found in biopsies from six individuals and positive Congo red staining after treatment with potassium permanganate was seen in biopsies from four of the cases. Biopsies of 100 patients suffering from non-amyloid renal disorders were also examined. None of them displayed any immunoreactive deposits of protein AA. The investigation shows that amyloid deposits of the AA-type can be identified in needle biopsies when sequence-specific antibodies against serum amyloid A are used in the avidin-biotin-peroxidase complex technique. Both the diagnostic sensitivity (42 of 42) and specificity (110 of 110) of the assay were optimal (1.0). The method was found to be superior to other investigated techniques and useful for classifying amyloidosis in formalin-fixed renal biopsies

Production of an amino acid sequence-specific antiserum against human amyloid A (AA) and serum amyloid A (SAA) protein

The hydrophilic nonapeptide Ser-Asp-Ala-Arg-Glu-Asn-Ile-Gln-Arg, identical with residues 59-67 of human amyloid protein A (AA) and serum amyloid protein A (SAA), was covalently bound via its carboxyl-terminal end to the carrier-protein keyhole limpet haemocyanin. The complex was injected subcutaneously into ten rabbits. All rabbits produced antisera which, unabsorbed, were specific for AA and SAA. The antisera and their isolated peptide specific antibodies were performance-tested and found to be excellent for demonstration of AA and SAA in immunoblotting and immunohistochemical techniques but unsuitable for immunoprecipitation. Since it is difficult to produce AA- and SAA-specific antisera by procedures earlier described and commercial supplies of good such reagents are unavailable, the easy production of sequence-specific such antisera will facilitate more extended studies of the corresponding antigens for diagnostic and scientific purposes