Hisayuki Inoue

Morphological and immunocytochemical identification of periacinar fibroblast-like cells derived from human pancreatic acini.

Fibroblast-like cells in the periacinar region may play an important role in periacinar fibrosis. In the present study, we isolated and cultured periacinar fibroblast-like cells (PFCs) derived from human pancreatic acini and examined the characteristics of human PFCs morphologically and immunocytochemically. Immunocytochemical study of human PFCs showed that they were positively stained with antibodies against type I collagen/procollagen, type III collagen/procollagen, fibronectin, prolyl hydroxylase β subunit, type IV collagen, laminin, α-smooth muscle actin, vimentin, and nonmuscle myosin. Electron microscopic study showed that human PFCs contained a number of microfilaments, forming dense bodies in the cytoplasm. These results indicated that human PFCs possess characteristics of myofibroblasts. Expression of α-smooth muscle actin, a marker of the myofibroblast-like phenotype, was increased with time in culture and was enhanced by treatment with transforming growth factor (TGF)-β1. Collagen synthesis in human PFCs was stimulated by TGF-β1 and the proliferation of human PFCs was stimulated by plateletderived growth factor. These findings suggest that PFCs from human pancreas seem to be involved in periacinar fibrosis.

Ulcerative colitis complicated by gastroduodenal lesions

A case of ulcerative colitis complicated with gastric and duodenal lesions is reported. The patient was a 17-year-old male who was admitted with bloody diarrhea and abdominal pain. Based on the endoscopic and histological findings of the colon, a diagnosis of ulcerative colitis was made. Upper gastrointestinal endoscopy showed multiple erosions and granular changes in the antral greater curvature of the stomach and descending portion of the duodenum. Histological examination of the stomach and duodenum revealed marked inflammatory cell infiltration and crypt abscesses. Clinically, the gastric and duodenal lesions did not respond to antiulcer drugs, but were alleviated by steroid. It was concluded that the pathogenesis of the gastric and duodenal lesions in this patient was similar to that of the colonic lesions of ulcerative colitis.

Effectiveness of combined anticoagulant therapy for extending portal vein thrombosis in Crohn's disease

PURPOSE: Portal vein thrombosis is a rare complication of Crohns disease, and its precise cause and appropriate treatment are not known. We describe a patient with extending portal vein thrombosis in Crohns disease who was successfully treated with combined anticoagulant therapy. METHOD: Urokinase and tissue plasminogen activator were administered from a catheter inserted into the superior mesenteric artery, and heparin and a serine protease inhibitor also were given intravenously. RESULTS: On admission, thromboembolic occlusion was observed throughout the entire portal venous system in association with massive ascites and remarkable intestinal edema. After administration of combined anticoagulant therapy, thrombus rapidly decreased in size, and color Doppler ultrasonography showed a gradual increase in portal venous flow. The patient had no recurrence of symptoms while receiving warfarin after resolution of thrombus. CONCLUSION: This case report suggests that combined anticoagulant therapy is effective for patients with severe portal vein thrombosis in Crohns disease and that color Doppler ultrasonography is useful for evaluation of portal venous flow.

Morphological identification of and collagen synthesis by periacinar fibroblastoid cells cultured from isolated rat pancreatic acini

Rat pancreatic periacinar fibroblastoid cells (PFCs) appear to be involved in intralobular fibrosis and acinar cell regeneration. We isolated pancreatic acini of the rat, cultured the fibroblastoid cells, and characterized the cells morphologically and immunohistochemically. Isolated acini were seeded on culture dishes, and spindle-shaped cells migrated and proliferated. On Electronmicroscopic examination, microfilament bundles were seen, and the intracellular localization of vimentin, α-smooth muscle actin, and non-muscle myosin was identified immunohistochemically. These findings strongly suggest that the cells were myofibroblast-like. The PFCs were also demonstrated, immunohistochemically, to contain prolyl hydroxylase, type-I procollagen, type-III procollagen, type-IV collagen, fibronectin, and laminin. Stimulation by transforming growth factor β1 (TGF β1) increased intracellular immunoreactive prolyl hydroxylase and collagen synthesis in the PFCs. These findings indicate that PFCs proliferate in culture as myofibroblast-like cells and synthesize extracellular matrix components. It is possible that PFCs are involved in intralobular fibrosis in response to stimulation with TGF β1.

Effects of serine protease inhibitors on accumulation of polymorphonuclear leukocytes in the lung induced by acute pancreatitis in rats

The administration of a high-dose of a serine protease inhibitor is recommended in patients complicated by multiple organ failure (MOF), including adult respiratory distress syndrome (ARDS), induced by acute pancreatitis. The accumulation of polymorphonuclear leukocytes (PMN) in affected organs is considered to be one of the causative factors of MOF. Adhesion to endothelial cells (EC), via adhesion molecules, and the transendothelial migration of PMN is closely associated with the accumulation of PMN. We examined the effects of two serine protease inhibitors, ulinastatin (UT) and gabexate mesilate (GM), on EC-PMN adhesion and transendothelial migration in human umbilical vein EC and51Cr-labeled PMN in vitro. EC-PMN adhesion, and the expression of intercellular adhesion molecule-1 (ICAM-1) and endothelial cell adhesion molecule-1 (ELAM-1) on EC induced by IL-1β and TNFα, were reduced by the pretreatment of EC with these inhibitors. The transendothelial migration of PMN stimulated by IL-8 was also inhibited by pretreating PMN with UT or GM. We also examined whether these inhibitors reduced PMN accumulation in the lung in rats with acute pancreatitis induced by a closed duodenal loop. The myeloperoxidase activity in and histological findings of the lung suggested that UT and GM reduced PMN accumulation. In conclusion, serine protease inhibitors may inhibit PMN accumulation in ARDS due to acute pancreatitis.

Acute lupus peritonitis successfully treated with steroid pulse therapy.

A 21-year-old man with systemic lupus erythematosus (SLE) who developed acute lupus peritonitis is described. Acute lupus peritonitis appeared during generalized lupus flare, with nausea, vomiting, frequent diarrhea, and abdominal tenderness with rebound and guarding. The patient was afebrile and had decreased bowel sounds. Abdominal ultrasonography and computed tomography revealed marked thickening of the gastric, duodenal, and jejunal walls, massive intraluminal fluid collection, and increasing ascites. Gastrointestinal endoscopy showed edematous mucosa with multiple erosions of the stomach and duodenum. The ascitic fluid was remarkable for low complement levels and elevated anti-DNA antibody. These manifestations of acute lupus peritonitis resolved after steroid pulse therapy with methylprednisolone. We should consider acute lupus peritonitis in a patient with SLE when abdominal symptoms are severe. Experience with our patient indicates that steroid pulse therapy is effective for this rare but severe manifestation of SLE.

Serum levels of interleukin-1β and interleukin-6 in patients with chronic pancreatitis

To investigate the role played by cytokines in chronic pancreatitis, we examined serum levels of interleukin-1β (IL-1β) and interleukin-6 (IL-6) by radioimmunoassay (RIA) and enzyme-linked immunosorbent assay (ELISA) in 33 patients with definitively diagnosed chronic pancreatitis. All the patients, who had received either no treatment or only digestive enzyme products for their chronic pancreatitis, had significantly elevated serum IL-1β levels (38.5±28.8pg/ml, mean ± SD), compared to normal controls (16.0±6.7pg/ml;P<0.01); however they showed no changes in serum IL-6 levels. Changes in IL-1β and IL-6 serum levels were not correlated with the etiological features of pancreatitis or with complications due to liver diseases. Serum IL-1β and IL-6 levels were also not correlated with the activity of any pancreatic enzymes in blood or urine. However, in the patients with chronic pancreatitis, serum IL-6 levels were correlated with C-reactive protein (CRP), whereas serum IL-1β levels were not correlated with CRP or with erythrocyte sedimentation rate. These results suggest that serum IL-1β is involved in the progression and reduction of chronic inflammation of the pancreas, and that the serum IL-1β level may be useful as a marker for chronic pancreastitis.

Macrolipasemia in Crohn's disease

A 38-year-old male patient who had been treated for Crohns disease was found to have serum lipase activity that was persistently increased - 10-fold above the normal upper limit. He was diagnosed with chronic pancreatitis based on slightly elevated elastase-1 level and retrograde pancreatography showing slight dilatation of the main pancreatic duct. Therefore, the hyperlipasemia was thought to be due to pancreatitis. However, the serum amylase and trypsin was not increased at any time, and no serious findings suggestive of pancreatitis were detected on morphologic examination. Thus, there were discrepancies between the serum lipase activity and other laboratory and clinical findings. Exclusion chromatography of the patients serum suggested macromolecular lipase, and further immunologic testing including affinity chromatography, enzyme-linked immunosorbent assay, and immunoprecipitation assay showed that serum lipase was bound to immunoglobulin GK. Therefore, the hyperlipasemia was caused by immunoglobulin-linked lipase, termed “macrolipasemia”. Macrolipasemia has rarely been reported, and this is the first reported case of macrolipasemia accompanied by Crohns disease.

Aggressive jejunal γδT-cell lymphoma derived from intraepithelial lymphocytes: An autopsy case report

Abstract: A 69-year-old man with massive ascites was referred to our hospital. Paracentesis revealed exudative ascites with many abnormal lymphocytes, which expressed T-natural killer (T-NK) cell surface markers and γδT-cell receptor (TCR). Although the ascites resolved for a short time with chemotherapy, gastrointestinal bleeding occurred and acute retention of ascites was observed. The patient died of panperitonitis. At autopsy, part of the jejunum revealed ulceration and marked mucosal thickening, and was perforated at the site of the severe ulceration. Histological examination showed massive infiltration of abnormal lymphocytes that were positive for CD45RO. Therefore, the cells responsible for the jejunal lymphoma and ascites were thought to be derived from γδIEL.

Effects of transforming growth factor α1, interleukin-1β, tumor necrosis factor α and platelet-derived growth factor on the collagen synthesis and the proliferation of periacinal fibroblastoid cells isolated and cultured from rat pancreatic acini

Abstract In vivo studies suggest that pancreatic periacinal fibroblasts participate in the development of pancreatic fibrosis, but regulatory mechanisms of their functions and proliferation have not been clarified. In this study, we examined the effects of humoral factors; transforming growth factor β1 (TGFβ1), interleukin-1β (IL-1β), tumor necrosis factor α (TNFα) and platelet-derived growth factor (PDGF), on the intracellular immunoreactive prolyl hydroxylase (IRPH) production, collagen synthesis and cell proliferation of periacinal fibroblastoid cells (PFC) isolated from rat pancreas. TGFβ1, IL-1β and TNFα induced a dose-dependent increase in IRPH production by PFCs. The most striking increase of intracellular IRPH was observed when PFCs were treated with 1.0 ng/ml of TGFβ1. TGFβ1 also significantly increased collagen synthesis at 1.0 ng/ml. Although, PDGF affected neither IRPH production nor collagen synthesis, it enhanced the cell proliferation of PFCs, dose-dependently. IL-1β and TNFα had no effect and TFβ1 rather inhibit the proliferation of PFCs. These findings suggest that both TGFβ1 and PDGF, synthesized locally from the infiltrated inflammatory cells, play important roles in the development of pancreatic fibrosis by increasing the collagen synthesis and enhancing the proliferation of PFCs, respectively, in vivo.