Serum concentration of N-terminal procollagen peptide of collagen type III in schistosomal liver fibrosis

The major cause of mortality in human schistosomiasis is the chronic granulomatous reaction of the liver tissue to Schistosoma mansoni eggs. Liver biopsy still provides the best evaluation of the degree of liver damage. However, liver biopsy does not provide an image of the dynamic process of fibrogenesis. Variations of concentrations of procollagen type III peptide in sera have been proposed to be significant markers of liver fibrosis. Thus, liver function tests in relation to histopathological diagnosis and procollagen type III peptide concentrations were studied in patients with schistosomiasis and revealed a high correlation between the serum procollagen type III peptide and the degree of fibrosis in liver tissue.     

Serum aminoterminal type III procollagen peptide. Relation to biosynthesis of collagen type III in experimentally induced granulation tissue in rats

Serum aminoterminal type III procollagen peptide was measured in rats during the development of granulation tissue induced by subcutaneous implantation of viscose cellulose sponges. Active collagen type III synthesis in granulation tissue during the first three weeks was accompanied by an increase in serum propeptide level. A positive correlation was observed between the increase in serum propeptide level on the one hand and the increase in granulation tissue collagen type III content and the in vitro formation of tissue 3H-hydroxyproline on the other hand. In some animals the serum propeptide level remained low, despite biochemical signs of collagen synthesis, indicating variations in the release into serum and/or the metabolism of circulating propeptide. The increase in propeptide antigen concentration was mainly due to an elevated content of material with molecular weight equal to or twice that of the propeptide. A minor fraction of the propeptide remained attached to the interstitial collagen fibres in the granulation tissue. The correlation between the serum propeptide level and the biosynthesis of collagen at the site of the focal fibroproliferative process suggests that the serum propeptide level may be a valuable indicator of fibrogenesis and thereby of disease activity in fibrotic conditions.     

Serum marker of type III procollagen in patients with idiopathic hemochromatosis and its relationship to hepatic fibrosis

A radioimmunoassay for serum procollagen III aminopeptide (sPIIIP) was proposed recently for monitoring hepatic fibroplasia in patients with various inflammatory hepatic lesions. To determine whether sPIIIP also can detect fibroplasia in noninflammatory liver disorders, we measured this index in 16 patients with idiopathic hemochromatosis (IHC) at various stages of the disease and iron overload. Interestingly, we found normal levels of sPIIIP in 12 out of 16 patients examined (75%), despite clear histologic features of fibrosis or cirrhosis. The levels of sPIIIP exhibited no relationship to any of the clinical, laboratory, or histologic parameters of the disease. Thus, unlike other types of cirrhosis, in which sPIIIP is increased, the liver disease in IHC may be a fibrotic process unrelated to type III collagen stimulation. Accordingly, the determination of sPIIIP in these patients is of no value for monitoring the fibrosis associated with the liver disease.     

Localization of type III procollagen mRNA in areas of liver fibrosis by in situ hybridization

Localization of type III procollagen mRNA in human liver was studied by in situ hybridization using human alpha 1(III) procollagen cDNA. Frozen and paraformaldehyde-fixed sections of biopsied human liver from patients with chronic hepatitis and cirrhosis were examined using the digoxigenin-labeled cDNA probe. Localization of the type III procollagen mRNA was demonstrated not only in the cytoplasm of mesenchymal cells but also in a large number of hepatocytes, in proportion to the extent of fibrosis. These results suggest that hepatocytes play an important role in fibrogenesis in the liver.     

Increased urine level of amino-terminal peptide derivatives of type III procollagen in patients with liver diseases

The amino-terminal peptides of type III procollagen (PIIIP) in the urine of 40 patients with various liver diseases were determined with a commercial radioimmunoassay kit. The level of urinary PIIIP (uPIIIP) was correlated well with serum PIIIP (sPIIIP) in 9 patients, the coefficient of correlation being r = 0.836 (p less than 0.01) and the regression line being y = 1.42x + 24. Urinary PIIIP consisted of at least 4 different molecular species with molecular weights of 49 k, 18 k, 10 k and 4.6 k as estimated by column chromatography on Sephadex G-100. Furthermore. uPIIIP was found to be significantly elevated in acute hepatitis, chronic hepatitis, liver cirrhosis, hepatocellular carcinoma and other liver diseases, in which the elevation of sPIIIP has been reported by others. The mean values +/- standard deviations of uPIIIP were 44.0 +/- 32.0, 60.4 +/- 32.0, 62.0 +/- 46.5, 53.0 +/- 27.1 and 48.1 +/- 22.8 ng/ml for the respective liver diseases, and 13.2 +/- 4.5 for the non-hepatic disease group.     

Serum asymmetric dimethylarginine levels are independently associated with procollagen III N-terminal peptide in nonalcoholic fatty liver disease patients

Although impaired synthesis and/or bioavailability of nitric oxide are considered to contribute to insulin resistance and the progression of liver disease in nonalcoholic fatty liver disease, role of asymmetric dimethylarginine, an endogenous inhibitor of nitric oxide synthase, has not been examined. We examined retrospectively which anthropometric and metabolic parameters were independently associated with serum levels of asymmetric dimethylarginine in nonalcoholic fatty liver disease. A total of 194 consecutive biopsy-proven nonalcoholic fatty liver disease patients with or without type 2 diabetes were enrolled. Serum asymmetric dimethylarginine levels in nonalcoholic fatty liver disease patients were significantly higher, irrespective of the presence or absence of diabetes, than those in healthy control. Multiple stepwise regression analysis showed that decreased total protein and procollagen N-terminal peptide levels, markers of advanced liver disease and hepatic fibrosis, respectively, were independently associated with asymmetric dimethylarginine levels in nonalcoholic fatty liver disease subjects without diabetes, whereas soluble form of receptor for advanced glycation end products and density ratio of liver to spleen in computed tomography were independent correlates of asymmetric dimethylarginine in diabetic patients. The present study suggests that asymmetric dimethylarginine may be associated with nonalcoholic fatty liver disease, especially subjects without diabetes.     

Serum enzymes of collagen synthesis and type III procollagen aminopropeptide in Nigerian patients with sickle cell disease

Serum immunoreactive prolyl hydroxylase protein (S-IRPH), galactosylhydroxylysyl glucosyltransferase activity (S-GGT), and the aminoterminal propeptide of type III procollagen (S-Pro(III)-N-P) were measured in 20 patients with sickle cell disease and the values were compared with those in 20 apparently healthy Nigerians. The means for the two enzymes and S-Pro(III)-N-P were significantly elevated in the sickle cell disease patients. Significant correlations were found between the values of the two enzymes and the protein (S-Pro(III)-N-P) within the sickle cell disease patients. The data confirm that collagen formation is found in the bone, liver, or other organs of patients with this disease. The measurement of S-GGT and S-Pro(III)-N-P in prospective studies might be helpful in predicting general and hepatic fibrogenesis in sickle cell disease.     

Study of serum procollagen type III peptide in patients with hepatic cirrhosis from a clinical point of view

Summary N-terminal procollagen-III peptide (P-III-P) has been considered a marker of fibrogenesis and inflammatory activity of the liver. We measured the P-III-P serum levels in 83 cirrhotic patients fully characterized from a clinical and laboratory point of view. Cirrhotic patients had significantly higher P-III-P serum levels than controls (P<0.0001). Of the cirrhotic patients 73.5% had increased P-III-P. A significant negative correlation was found between P-III-P and transaminases, and patients with normal values of alanine aminotransferase had higher P-III-P serum levels than those with increased values (P = 0.03). On the other hand, no significant association was found with portal hypertension, Child classes, or alcoholic liver disease. No one independent factor appears to be responsible for the increase in P-III-P. The measurement of serum P-III-P is of little if any use in the evaluation of cirrhotic patients.Abbreviations P-III-P procollagen-III peptide - ALT alanine aminotransferase     

Ultrastructural localization of type III procollagen in baboon liver.

The localization of the extension aminopropeptides of Type III procollagen was studied in baboon liver by the immunoperoxidase method. The aminopropeptides were localized along the collagen fibrils around terminal hepatic venules, in the space of Disse, and in portal tracts of control baboons as well as of alcohol-fed animals. The labeling showed a characteristic periodicity of 60-70 nm, corresponding to the D (67 nm) stagger of collagen fibrils. The data indicate that some extension aminopropeptides of Type III procollagen are not cleaved from procollagen prior to its incorporation into collagen fibrils but that they exist as constituents of mature fibrils.     

Differential patterns of PMN-elastase and type III procollagen peptide in knee joint effusions due to acute and chronic sports injuries

Summary In 38 traumatic knee joint effusions the proteolytic enzyme PMN-elastase (PMN-E) and the repair marker procollagen III aminoterminal peptide (PIIINP) were determined. According to the period between trauma and first aspiration of the effusion, the patients were divided into 3 groups. Group I (17 patients; period between trauma and first aspiration not longer than 72 hours) showed high concentrations of PMN-E (up to 5400 ng/ml) and low concentrations of PIIINP (<13 U/ml). Group II (11 patients; aspiration within 4 to 14 days) had mean PMN-E and PIIINP concentrations of 125.6 ng/ml and 52.1 U/ ml, respectively. In group III (10 patients, aspiration after 14 days) mean PMN-E concentration was 123.8 ng/ml and mean PIIINP concentration was 63.4 U/ml. Graphic depiction of PMN-E and PIIINP levels in each individual sample as a function of time between trauma and fluid collection revealed highly increasing PMN-E levels during the first 24 posttraumatic hours, followed by rapidly decreasing levels within 72 hours post trauma, and no change after the 4th posttraumatic day. In contrast, PIIINP increased continuously up to the first posttraumatic week and stayed at high levels up to 90 days (end of the observation period). The differential patterns of PMN-E and PIIINP concentration in knee joint effusions may be useful in estimating the period between trauma and first treatment (aspiration of effusion) and should, therefore, be helpful in detecting degenerative lesions, which seem to be characterized by low PMN-E concomitantly with high PIIINP levels.Abbreviations PMN-E elastase from polymorphonuclear granulocytes - PIIINP procollagen III aminoterminal peptide - 1PI 1-proteinase inhibitor     

Ehlers-Danlos syndrome IV due to a novel defect in type III procollagen

Ehlers-Danlos syndrome type IV (EDS IV) is characterized by variable changes in the skin, arterial fragility, bowel perforation, minimal joint involvement, and either autosomal recessive or autosomal dominant inheritance. The unifying biochemical abnormality is a deficiency of type III collagen; all patients studied thus far have shown a defect in either synthesis or in secretion of type III procollagen. We report on an adolescent boy who inherited EDS IV from his father and who developed a spontaneous subclavian artery aneurysm. In vitro studies of collagen production in dermal fibroblasts showed normal amounts of pro α1 (III) messenger RNA and synthesis and secretion of nearly equal amounts of normal and of structurally abnormal pro α1(III) monomers. This patient is biochemically distinct from previous cases of EDS IV and is likely heterozygous for a mutation that results in an aberrant type III procollagen that is particularly susceptible to protease degradation.     

Immunohistochemical and immunoelectron microscopical localization of procollagen III peptide in papillary carcinoma of thyroid

The immunohistochemical localization of procollagen III peptide, a precursor of type III collagen, was examined in 38 papillary carcinomas and 25 follicular neoplasms using an immu-noperoxidase technique. Although localization of procollagen ill peptide was not demonstrated in normal follicular cells, distinct cytoplasmic immunostaining of neoplastic cells was frequently found in the tissues examined, as were stromal fibroblasts. Such cytoplasmic immunostaining was observed in 92.1 % of 38 papillary carcinomas and in 36.0% of 25 follicular neoplasms. Cytoplasmic immunoreactivity in papillary carcinomas was more intense at the peripheral zone of the tumor and correlated with the degree of invasiveness. The controls, in which the primary antibody was preabsorbed with procollagen ill peptide or replaced with normal rabbit serum, showed only faint background staining in all specimens. Immunoelectron microscopical examination revealed that electron-dense deposits, indicating procollagen III peptide, were located in the perinuclear space, Golgi apparatus, and rough endoplasmic reticulum of papillary carcinoma cells. These results suggest that neoplastic cells, especially papillary carcinoma cells, could play an important role in type III collagen production, possibly in connection with the creation of an environment that is conducive to their progression.     

Determination of the aminoterminal peptide of procollagen type III and laminin P1 in serum of patients with chronic liver disease

Serum concentration of the aminoterminal peptide of procollagen type III (P III P) and that of the high-molecular-weight glycoprotein laminin P1 (LP1) were determined by a specific radioimmunoassay (RIA) in patients with different chronic liver diseases. Besides the routine laboratory tests, histological verification of the liver samples obtained by needle biopsy and a complex hepatitis B virus marker analysis by RIA (Biomedica-Sorin), or ELISA (Behringwerke, Marburg, FRG) kits were carried out in order to set up the correct clinical diagnosis. In normal controls, the P III P and LP1 concentrations were 7.8 +/- 1.1 ng/ml (n = 10) and 0.08 +/- 0.1 units/ml (n = 7), respectively. Patients with fatty liver (n = 25) showed a significant elevation in P III P concentration (18.6 +/- 2.7 ng/ml). Such an elevation was not unequivocally demonstrated before. In this group of patients LP1 level was also increased (1.4 +/- 0.2 units/ml, n = 10). In liver cirrhosis (n = 51) both P III P and LP1 concentrations were found to be consistently elevated.     

Localization of type III procollagen aminopeptide antigenicity in hepatocytes from cirrhotic human liver

Summary Immunolocalization of type III procollagen (pro III) in normal and cirrhotic human liver was studied using rabbit antiserum specific for bovine type III procollagen aminopeptide. The material examined was deparaffinized, trypsin-treated hepatic tissue sections from 28 autopsy cases, including 19 cirrhotic and 9 normal liver donors. Immunostaining, performed by the unlabeled peroxidase-antiperoxidase antibody technique demonstrated that extracellular matrices corresponding to perisinusoidal reticulin, collagen in periportal areas, and blood vessel walls were the common sites of pro III antigenicity in both normal and cirrhotic liver. Moreover, in the cirrhotic liver, the fibrous septa of pseudolobules, and cytoplasm of hepatocytes and sinusoidal cells were positive when stained for pro III peptide. The differential counts of pro III positive cells in cirrhotic liver, however, revealed that the average ratio of these hepatocytes to sinusoidal cells was 25 to 1, indicating complete dominance of hepatocytes with respect to stainability for pro III peptide compared to sinusoidal cells. In hepatocellular carcinomas co-existing with cirrhosis, neoplastic cells also displayed pro III antigenicity.These data suggest that hepatocytes of cirrhotic liver and hepatocellular carcinoma cells play a significant role in type III collagen synthesis in vivo.The paper on this problem was read at the International Symposium on Pathobiology of Hepatic Fibrosis held in Matsue, Japan, on June 16, 1985     

Immunolocalization of type III collagen and procollagen in cirrhotic human liver using monoclonal antibodies

Summary Immunolocalization of type III collagen and procollagen in cirrhotic human liver was studied using monoclonal antibody specific for the helical determinant of type III collagen extracted from human placenta. Deparaffinized, trypsin-treated cirrhotic liver sections from 8 autopsy cases were examined by the unlabeled peroxidase-antiperoxidase and immunofluorescence techniques. These techniques revealed the localization of this epitope shared by type III collagen and procollagen not only in the extracellular matrix of hepatocytes and sinusoidal cells but also in the cytoplasm. In hepatocellular carcinoma concurrent with cirrhosis, neoplastic cells were shown to react with this antibody as well.These results are consistent with data obtained using antiserum specific for bovine type III procollagen aminopeptide which appeared in our previous report.     

Involvement of collagen-binding heat shock protein 47 and procollagen type I synthesis in idiopathic pulmonary fibrosis: contribution of type II pneumocytes to fibrosis

The most common pathologic form of idiopathic pulmonary fibrosis is usual interstitial pneumonia, which is characterized by patchy fibrotic areas, marked increase in the number of fibroblasts and type II pneumocytes, and excessive deposition of extracellular matrix proteins, especially collagen. Heat shock protein 47 is a collagen-binding stress protein and has a specific role in intracellular processing of procollagen molecules as a collagen-specific molecular chaperone. However, its role in the causation of fibrosis in usual interstitial pneumonia is unknown. In this study, we examined the expression of heat shock protein 47 and type I procollagen in 12 patients with usual interstitial pneumonia by immunohistochemistry on sequential sections. Heat shock protein 47 was localized predominantly in alpha-smooth muscle actin-positive myofibroblasts and surfactant protein-A-positive type II pneumocytes in active fibrotic areas of usual interstitial pneumonia. Type I procollagen was also expressed in those cells. In contrast, heat shock protein 47 and type I procollagen were weakly or not at all expressed in myofibroblasts and type II pneumocytes in bronchiolitis obliterans organizing pneumonia and normal lung tissue samples obtained from excised lung cancer tissues. The numbers of heat shock protein 47- and type I procollagen-positive cells to type II pneumocytes or myofibroblasts were significantly higher in usual interstitial pneumonia than in bronchiolitis obliterans organizing pneumonia and normal lung tissue specimens. Our results suggest that myofibroblasts and type II pneumocytes play an important role in the progression of fibrosis through the induction of heat shock protein 47, which regulates the synthesis/assembly of type I procollagen in usual interstitial pneumonia. HUM PATHOL 31:1498-1505.     

实验性肝纤维化中I,III型胶原的形态学观察

Rats receiving intravenous injection of human albumin (4 mg/rat) once biweekly developed liver fibrosis. The lesion seemed to have little relation to hepatocellular injury. The incidence of liver fibrosis increased with the length of immunization, ranging 80%-86% after 30-60 days. The whole process of experimental liver fibrosis may be divided into three phases. The first is the initial stage of fibroplasia (from the first day to the time of 15 days after start of immunization). In this phase, Ito cells were activated and collagen type I and type III began to increase. The second is the activated stage of fibroplasia (from the 15th to the 60th day after the beginning of immunization), in which collagen type I and type III reached the maximum and myofibroblasts as well as 'transitional cells' proliferated with deposition of collagen fibers. The third phase is the stage of post-fibroplasia (the period after elimination of immunization). Collagen type III diminished gradually while collagen type I remained increasing. Our findings indicated that fibroplasia occurs at the very beginning of liver injury. This suggest that treatment of liver fibrosis must be considered simultaneously with treatment of acute hepatitis.     

Altered secretion of type III procollagen in a form of type IV Ehlers-Danlos syndrome. Biochemical studies in cultured fibroblasts

Cultured dermal fibroblasts from a woman with one variety of type IV Ehlers-Danlos syndrome synthesized type III procollagen but fail to secrete the bulk of the protein. Although total collagen production is similar to that of controls, the affected cells retain almost twice as much collagen as controls. The additional retained protein is a disulfide-boned collagenous trimer that remains disulfide-linked after limited proteolysis with pepsin and, after pepsin treatment, migrates with type III collagen on polyacrylamide gel electrophoresis. Affected cells have markedly increased staining with antibodies directed against type III procollagen. These studies indicate that decreased secretion of type III procollagen that is synthesized can result in the clinical syndrome of type IV Ehlers-Danlos syndrome.