Effect of an immunotoxin to folate receptor beta on bleomycin-induced experimental pulmonary fibrosis.

It has been suggested that alveolar and interstitial macrophages play a key role in the pathogenesis of idiopathic pulmonary fibrosis (IPF) by producing proinflammatory and/or fibrogenic cytokines. We showed that inflammatory macrophages expressed folate receptor beta (FRbeta) while resident macrophages in normal tissues expressed no or low levels of FRbeta. In the present study, we examined the distribution of FRbeta-expressing macrophages in the lungs of patients with usual idiopathic pulmonary fibrosis (UIP) and mice with bleomycin-induced pulmonary fibrosis (PF) and tested whether the depletion of FRbeta-expressing macrophages could suppress bleomycin-induced PF in mice. Immunostaining with anti-human or -mouse FRbeta monoclonal antibody (mAb) revealed that FRbeta-expressing macrophages were present predominantly in fibrotic areas of the lungs of patients with UIP and mice with bleomycin-induced PF. Intranasal administration of a recombinant immunotoxin, consisting of immunoglobulin heavy and light chain Fv portions of an anti-mouse FRbeta mAb and truncated Pseudomonas exotoxin A, increased survival significantly and reduced levels of total hydroxyproline and fibrosis in bleomycin-induced PF. In immunohistochemical analysis, decreased numbers of tumour necrosis factor-alpha-, chemokines CCL2- and CCL12-producing cells were observed in the immunotoxin-treated group. These findings suggest a pathogenic role of FRbeta-expressing macrophages in IPF. Thus, targeting FRbeta-expressing macrophages may be a promising treatment of IPF.

Expansion of a unique macrophage subset in rheumatoid arthritis synovial lining layer.

The Z39Ig protein (complement receptor for C3b and iC3b) is expressed on resident tissue macrophages in various tissues. This study was undertaken to examine the distribution of Z39Ig+cells and their phenotypic features in rheumatoid arthritis (RA) synovium, in comparison with those of osteoarthritis (OA) and psoriatic arthritis (PsA) synovium. Monoclonal anti-Z39Ig antibody was produced by immunizing Z39Ig transfected murine pre B cells and used for the identification of Z39Ig+cells. Z39Ig+cells were further stained with antibodies to macrophages, fibroblast-like synoviocytes, complement receptors and dendritic cells by using the double immunostaining method in normal, RA, OA and PsA synovium. RA synovial mononuclear cells were double-stained using anti-Z39Ig and anti-CD11c antibodies and sorted into Z39Ig+CD11c+cells and Z39Ig+CD11c-cells. These cell populations were then analysed by electron microscopy. The expression of the Z39Ig protein was limited to intimal macrophages in normal, RA, OA and PsA synovium. The numbers of Z39Ig+CD11c+cells and the ratios of Z39Ig+CD11c+cells to Z39Ig+cells were increased in the synovial lining layer of RA as compared with those of OA and PsA. The ultrastructural analysis of Z39Ig+CD11c+cells showed the character of macrophages with many secondary lysosomes and swelling of mitochondria. Z39Ig+ cells appeared to be useful for identification of resident tissue macrophages in normal synovium and the corresponding macrophages in the synovial lining layer of inflammatory arthritis. Expansion of Z39Ig+CD11c+cells was characteristic of RA synovial lining layer.

Idiopathic severe pulmonary hypertension in an infant with pulmonary infection.

We report a 5-month-old infant who showed typical echocardiographic findings of primary pulmonary hypertension without the typical histopathological findings and who recovered from severe pulmonary hypertension. Histopathological findings revealed mild thickening of small pulmonary arteries and activated macrophages in the lung. Some cases with idiopathic severe pulmonary hypertension in infants are associated with pulmonary infection.

Comparative study for histology, proliferative activity, glycoproteins, and p53 protein between old and recent colorectal adenomas in Japan.

The incidence of colorectal carcinoma is increasing in Japan. Malignant transformation in colorectal neoplasia is usually considered to be owing to adenoma-carcinoma sequence. Elucidation of the recent alteration in the biological properties of colorectal adenoma is sure to be useful to understand the recent increase of the colorectal carcinoma in Japan. We compared the histopathological feature, mitotic index, proliferative activity (Ki-67 labeling index), expression of glycoproteins such as MUC2 mucin, sialyl Lewis A (SLe(a)) and sialyl dimeric Lewis X (SLe(x)), and p53 protein overexpression, between 108 adenomas in the old period (Group A, from 1969 to 1985) and 140 adenomas in the recent period (Group B, from 1995 to 1998). The histological dysplasia, mitotic index and Ki-67 labeling index of the adenomas were significantly higher in Group B than in Group A. In contrast, the expression of MUC2 mucin, which is considered to be a differentiation factor of intestinal mucosal epithelium, was significantly reduced in Group B than in Group A. The SLe(a) and SLe(x) expressions showed no significant difference between them. The p53 expression showed no significant difference between them, except for the moderate dysplasia. These findings indicate that recent colorectal adenomas show more advanced degrees of histological dysplasia, more rapid growth, and reduced differentiation than colorectal adenomas, which developed at earlier times, and may be related with the recent high incidence of colorectal carcinoma in Japan.

Expression of epithelial growth factor receptor and its two ligands, transforming growth factor-alpha and epithelial growth factor, in normal and neoplastic squamous cells in the vulva: an immunohistochemical study.

Epithelial growth factor receptor (EGFR) sends signals to the proliferation signal transduction system, receiving two ligands: epithelial growth factor (EGF) and transforming growth factor-alpha (TGF-alpha). This immunohistochemical study examined the roles of EGFR and its ligands in the proliferation of normal and neoplastic vulvar squamous cells in 25 patients with vulvar squamous cell carcinoma (VSCC), 10 patients with vulvar condyloma acuminata (VCA), 15 patients with vulvar intra-epithelial neoplasm I-II or III (VIN I-II or III), and 5 subjects with vulvar normal squamous cells (VNSC). EGFR was detected in a few basal cells in 40% of the VNSC, in highly dysplastic cells in 40% of the VIN III, in many neoplastic cells in 80% of the VCA, and in some malignant cells in 64% of the VSCC. EGF was seen in the cytoplasm in 20% of the VIN I-II, 100% of the VIN III, 100% of the VCA, and 100% of the VSCC. Diffuse TGF-alpha was weakly expressed in the cytoplasm in 100% of the VNSC, more intensely in 100% of the VIN and 100% of the VCA, and intensely in 100% of the VSCC. These findings led to the suggestion that the TGF-alpha-EGFR system maintains the growth of normal squamous cells and, in part, maintains the growth of dysplastic and neoplastic squamous cells in the vulva. EGF expression was an early sign of neoplasia. The expression of EGFR with overexpression of its two ligands contributed to the proliferation of dysplastic and neoplastic squamous cells in VIN III and VCA. EGFR expression appeared to contribute to essential neoplastic abnormalities in 64% of the VSCC.

Immunohistochemical estimation of in-situ cell cycle time in neoplastic epithelial cells in human large intestine: a new cell proliferation index.

In-situ cell cycle time (in-situ Tc) of epithelial cells could be estimated by using a formula; in-situ Tc = cell proliferation rate divided by mitosis rate, on a scale of Tm (cell cycle time in M phase) arbitrary unit (AU), In order to see the nature of in-situ Tc in the adenoma-carcinoma sequence in the human large intestine, the in-situ Tc in 27 cases of adenoma and 71 cases of adenocarcinoma with adenoma components in the human large intestine was estimated by using this formula, counting proliferating cells and mitotic cells in the immunohistochemistry of Ki-67 antigen. C12 antigen was examined as an oncogenic progression indicator in the adenoma-carcinoma sequence. The in-situ Tc tended to shorten in adenoma in accordance with the histological grading of atypia but not in adenoma component. No significant differences in the in-situ Tc was recognized as a whole among adenomas, adenoma components and adenocarcinomas in the mucosa, whereas the in-situ Tc of adenoma components with moderate to severe atypia was significantly longer than that of adenocarcinomas in the mucosa (p = 0.05). The in-situ Tc lengthened in adenocarcinomas invading the submucosa and shortened in adenocarcinomas invading the proper muscular layer. The cases expressing the C12 antigen increased in order of adenoma, adenoma component and adenocarcinoma. The cases expressing the C12 antigen indicated short in situ Tc in the adenomas and adenocarcinomas but not in the adenoma components. Thus, the estimated in-situ Tc is a useful index of the oncogenetic progression, which is different from that detected by the C12 antigen.

Immunohistochemical analysis of pericryptal fibroblast sheath and proliferating epithelial cells in human colorectal adenomas and carcinomas with adenoma components.

In order to examine stromal-epithelial interaction during the oncogenic progression of large bowel tumors, the association between pericryptal fibroblast sheath (PCFS) and expression of Ki-67 antigen was evaluated in 87 cases of colorectal adenoma and 95 cases of carcinoma with an adenoma component (CWA). For the immunohistochemistry, anti-alpha-smooth muscle actin antibody (alpha-SMA) and anti-Ki-67 antigen antibody (MIB-1) were used. In adenomas and adenoma components of CWA, the quantity of neoplastic glands with PCFS was reduced relative to the progression of histological atypia. Pericryptal fibroblast sheath was virtually absent from invasive carcinoma areas of CWA. Increased expression of Ki-67 correlated with the degree of histological atypia of adenomas. A significant reverse correlation was also seen between Ki-67 expression and PCFS-positive glands in adenoma components of CWA. These findings suggest that the prevalence of PCFS and Ki-67 expression are important indicators of colorectal neoplasia progression. The significant reduction of PCFS in colorectal epithelial neoplasms reflects progression in the adenoma-carcinoma sequence.

Molecular pathologic analysis of the tonsil in HTLV-I-infected individuals.

Little is known about the role of the tonsils in HTLV-I infection. We performed molecular pathologic studies of tonsils in individuals positive or negative for anti-HTLV-I antibodies (HTLV-I-Ab) to clarify histologic characteristics of tonsils in HTLV-I infection. We collected tonsils and peripheral blood samples from patients who underwent tonsillectomy in a prospective manner. HTLV-I-Ab in serum was examined and presence of HTLV-I provirus was detected by polymerase chain reaction (PCR) in extracted DNA of both peripheral blood and tonsils. Histopathologic and immunohistochemical evaluations of tonsils were performed. HTLV-I seropositivity and PCR detection of HTLV-I provirus matched perfectly. Tonsil samples from seropositive individuals showed atrophy of the mantle zone and high numbers of T cells in the marginal zone compared with findings in HTLV-I-negative samples. HTLV-I provirus could be detected only from extracted DNA of extrafollicular areas. PCR in situ hybridization also showed positive signals in some mononuclear cells located in the marginal zone. There was a significant correlation between HTLV-I proviral load in tonsils and in peripheral blood. These results suggest the presence of characteristic histologic changes and deviated localization of HTLV-I-infected cells in the tonsils of individuals positive for HTLV-I.

Protein-losing enteropathy due to thrombophlebitis of the mesenteric vein: report of a case.

We report herein the case of a 70-year-old woman with enteropathy accompanied by protein loss, the cause of which was found to be thrombophlebitis of the mesenteric vein. The patient was admitted to our hospital for investigations to determine the cause of hypoproteinemia. She had suffered an episode of left abdominal pain with high fever and vomiting lasting 10 days, 8 months prior to her admission. She also had a 6-year history of uncontrolled diabetes. The alpha1-antitrypsin clearance was 85.7 ml/day, suggesting protein-losing enteropathy. A scintigraphy with 99m-technetium-human serum albumin disclosed protein leakage into the intestine. X-Rays and computed tomography showed a stenotic and thickened area of small intestine 50 cm in length. Thus, a laparotomy was performed to resect this part of the intestine which was found to have undergone past thrombophlebitic changes. Following the operation, the alpha1-antitrypsin clearance decreased to within the normal range and the patient gained 5 kg in weight.

Protein kinase C has two different major roles in lattice compaction enhanced by cerebrospinal fluid from patients with subarachnoid hemorrhage.

BACKGROUND AND PURPOSE: Compaction of extracellular matrix (ECM) lattices by cultured fibroblasts was accelerated by cerebrospinal fluid (CSF) from patients with subarachnoid hemorrhage (SAH). The rate of acceleration was significantly related to the clinical grade of vasospasm. However, the mechanism remains unclear. Evidence exists for an important role in cerebral vasospasm for protein kinase C (PKC). The purpose of this study was to help clarify whether PKC has a role in contraction of the ECM. METHODS: We studied the effects of a myristoylated PKC peptide inhibitor (Myr-Arg-Phe-Ala-Arg-Lys-Gly-Ala-Leu-Arg-Gln-Lys-Asn-Val) (PKC peptide inhibitor), (5-isoquinolinesulfonyl)-homopiperazine (HA-1077) (inhibitor of protein kinase A, myosin light-chain kinase, and protein kinase G), 7-deacetyl-6-(N-ace-tylglycyl)-forskolin (forskolin) (adenyl cyclase activator), and diacylglycerol-lactone (DAG-lactone) (PKC activator) on fibroblast-populated collagen lattice compaction with or without CSF from SAH patients. Four sets of fibroblasts were used: three explanted from skin and one from cerebral artery. RESULTS: Moderate and high concentrations of PKC peptide inhibitor inhibited lattice compaction with or without acceleration by CSF. Low concentration of PKC peptide inhibitor enhanced acceleration by CSF but had no effects without CSF. HA-1077 could not inhibit lattice compaction. Forskolin inhibited compaction. DAG-lactone accelerated compaction in early phases. CONCLUSIONS: In the mechanism of acceleration of contraction of ECM under the influence of CSF, PKC seems to have two different roles. Protein kinase A and myosin light-chain kinase apparently play more minor roles than PKC in the mechanism, but no evidence was found of a role for protein kinase G activation in matrix compaction.

Effects of NG-nitro-L-arginine on isolated rabbit afferent arterioles.

We examined the effects of NG-nitro-L-arginine (L-NNA) on isolated rabbit afferent arterioles to confirm that nitric oxide is released at the resistance vessel level in the kidney. We microdissected the superficial afferent arterioles from the kidneys of New Zealand White rabbits. Each afferent arteriole was cannulated with a micropipette system, and the intraluminal pressure was set at 80 mmHg. By our methods, we found that norepinephrine (NE) decreased the lumen diameter of the afferent arterioles in a dose-dependent manner, and acetylcholine increased the lumen diameter of NE-constricted afferent arterioles. L-NNA (10(-4) M) gradually decreased the lumen diameter of afferent arterioles from 21.5 +/- 0.9 to 18.6 +/- 0.9 microns in 20 min, but NG-nitro-D-arginine (10(-4) M) did not affect them (from 21.8 +/- 1.3 to 21.8 +/- 1.5 microns). L-Arginine (10(-2) M) restored the lumen diameter of L-NNA-contracted afferent arterioles to the control levels. These findings indicate that the isolated afferent arteriole has the ability to release or to synthesize and release nitric oxide under basal conditions and that this basal release of nitric oxide plays an important role in the basal tone of the afferent arteriole.

Inflammatory pseudotumor of the spleen: report of a case.

A case of an inflammatory pseudotumor arising in the spleen of a 60-year-old Japanese male is described herein. This benign lesion is extremely rare, with only 12 cases, including our own, having been reported in the world literature. We preoperatively diagnosed the splenic tumor as a metastasis, due to the coexistance of advanced stage carcinoma in the sigmoid colon. However, after splenectomy, histopathological examination of the mass revealed an inflammatory process. Inflammatory pseudotumors often pose diagnostic difficulties because the clinical and radiologic findings are suggestive of malignancy. The clinical and pathological features of cases previously reported are reviewed following the presentation of this case.

Acute abdomen due to adenomyosis of the uterus: a case report.

A Japanese woman with severe abdominal pain underwent an emergent exploratory laparotomy. Intraperitoneal bleeding from the uterine serosal surface was observed, and a pathologic examination showed that the bleeding was caused by an exudative hemorrhage from the endometrial tissues in the myometrium close to the uterine serosal surface.

Is there any effect of volcanic eruptions of Mount Sakurajima on canine lungs exposed naturally?--Morphometric analysis of intrapulmonary particulate deposit amount and histopathological investigations.

In order to see whether any effect of inhalation of volcanic ash and gases from Mt. Sakurajima on canine lungs is observed or not, we examined the amount of intrapulmonary particulate deposits (IPD) and histopathological changes. Twenty-five abandoned or stray dogs (group A) in the areas affected enormously by volcanic ash and gases were examined in comparison with 13 abandoned or stray dogs (group B) in the area scarcely influenced. The amount of IPD was measured by using an image analyzer combined with a microscope. Age-associated increase of IPD values was noted, but mean IPD values were not different between groups A and B. Incidence of goblet cell hyperplasia was not different between the two groups. In none of the cases examined, squamous metaplasia of respiratory epithelia, pulmonary fibrosis, silicotic nodules, emphysematous change, or histopathological findings, which are indicative of bronchial asthma, were observed. In conclusion, obvious effect of volcanic eruption on canine lungs was not observed through both the measurement of IPD value and the histopathological evaluation.

Immunohistological quantitative analysis of S100 protein-positive cells in T-cell malignant lymphomas, especially in adult T-cell leukemia/lymphomas.

S100 protein-positive cells (S100+ cells) in 36 cases of T-cell lymphoma (T-ML) in the lymph node and 15 cases of T-ML in the skin were analyzed immunohistologically in order to study their quantitative features in adult T-cell leukemia/lymphoma (ATLL). The T-MLs were categorized according to the updated Kiel classification, and the T-cell pleomorphic type (Pleo) was subcategorized into 3 subtypes: Pleo-ATLL, Pleo-clear and Pleo-others. The population of S100+ cells and the first to fifth minimal distances of every S100+ cell were measured on micrographs of paraffin sections that had reacted to anti-S100 protein antibody according to the ABC method. Lymphoblastic and chronic lymphocytic leukemia types showed low populations of S100+ cells and long values of the first minimal distance. T-zone lymphoma without follicles and angioimmunoblastic lymphadenopathy with dysproteinemia-type T-ML had high populations and low values of the first minimal distance. Among the three subtypes of Pleo in the lymph node, Pleo-ATLL gave the highest population and the shortest value of the first minimal distance of S100+ cells, but this trend was not found in the skin. Clusters of more than five S100+ cells were more common in the Pleo-ATLL subtype than in the other two subtypes. The increase and clustering of S100+ cells in Pleo-ATLL suggests that the lymphoma cells act on S100+ cells as a helper.(ABSTRACT TRUNCATED AT 250 WORDS)

Chromaticity analysis of immunostained tumor specimens.

In order to evaluate the correlation between immunohistochemical and morphometric data on the same histological sections, we have developed a flexible color image analyzer (Microcomputer-Assisted Picture Processing System type II, MAPPS-II), and established an effective method to analyze the immunostained colorectal neoplasms based on the color recognition theory of human visual system. Colorectal adenomas and adenocarcinomas were stained with a monoclonal antibody C 12, which recognizes abnormal H antigen, using Avidin-Biotin method and diaminobenzidine (DAB, brown dye). Nuclei were stained with Hematoxylin (blue dye). Density and colorimetric analyses revealed two results: (A) Separation of immunostained brown area from blue nuclei was best performed by plotting the representative sample areas on a standard chromaticity diagram, which displays the hue and saturation of colors simulating color of human visual system. (B) After separation of immunostained areas, usual density analysis was useful for the assay of nuclear morphometric information. Using these programs, normal mucosa was negative for C 12, and showed low nuclear/cytoplasmic ratio (NCR). Adenoma was occasionally focally positive for C 12, and showed medium NCR. Carcinomas were C 12 positive, and showed high NCR. Our method permits nuclear counterstaining by hematoxylin instead of low contrast methyl green, which will widen the field of combined immunohistochemical and morphometric study.

Immunoblastic lymphadenopathy (IBL)-like T-cell lymphoma in a child.

We report the case of a 14-year-old Japanese boy with peripheral T-cell malignant lymphoma, showing progression from immunoblastic lymphadenopathy (IBL) to overt malignant lymphoma. He suffered recurrent fever, generalized lymphadenopathy, hepatosplenomegaly and maculopapular exanthema. Leukocytosis with eosinophilia and polyclonal hypergammaglobulinemia were observed during the aggressive course of the disease. In the early phase, human immunoglobulin and steroids improved the symptoms but did not induce complete remission, and the patient died one year after the onset of the illness. Four biopsies of lymph nodes revealed progression from IBL to CD4 positive T-cell lymphoma through IBL-like T-cell lymphoma. Though IBL-like T-cell lymphoma is defined as IBL with neoplastic features and overt T-cell malignant lymphoma progressed from IBL-like T-cell lymphoma is excluded from the definition, it may be preferable that such malignant lymphoma as our case should also be included in IBL-like T-cell lymphoma.

Paraffin-immunohistochemical analysis of 226 non-Hodgkin's malignant lymphomas in the endemic area of human T-cell leukemia virus type 1.

A study was conducted to evaluate the usefulness of paraffin-immunohistochemistry for histopathological classification of non-Hodgkin's malignant lymphomas (NHML). the phenotypes of lymphoma cells and other cells were examined using 11 monoclonal and 3 polyclonal antibodies by the ABC method on paraffin-embedded tissue sections of 226 cases of NHML, comprising 94 B-cell lymphomas (B-ML) and 132 T-cell lymphomas (T-ML). In 219 NHML cases (96.8%), lymphoma cells reacted with more than one of these antibodies. A set of MB-1, Mx-pan B, L26, LN-1, LN-2 and anti-immunoglobulin light chain antibodies characterized each subtype of B-MLs, categorized according to the Kiel classification. Mantle-zone lymphoma (MzML) was added as one subtype. L26 stained the largest number of B-MLs (82.8%). B-cell chronic lymphocytic leukemia (B-CLL) was labeled most frequently by MB-1. MzML was characterized by reactivity of lymphoma cells with LN-2 and by the appearance of monoclonal immunoglobulin light chain along the cell membrane. Follicle center cell lymphomas were stained by LN-1 and LN-2, although a small number of proliferating cells were labeled by LN-1 in B-CLL, MzML and the immunocytoma lymphoplasmacytic/cytoid variant. MT-1 and/or UCHL-1 showed various degrees of reactivity with the cell membranes of lymphoma cells in 94.8% of T-MLs. Among the T-cell pleomorphic lymphomas of Suchi and Lennert, the adult T-cell leukemia/lymphoma type, defined by stippled heterochromatin distribution and peculiar huge cells, reacted selectively (p less than 0.05) with anti-phosphokinase C antibody. Anaplastic large cell T-ML reacted with a set of Ber H2, LN-2 and Leu M1. In T-zone lymphomas without hyperplastic follicles, angioimmunoblastic lymphadenopathy with dysproteinemia-type T-ML, lymphoepithelioid cell lymphomas and some pleomorphic lymphomas comprising clear large lymphoma cells, there were many intermingling B cells, and their constitution varied. In some lymphoblastic lymphomas of both the T cell and B-cell type, phenotypes of T cells and B cells were expressed. Consequently, it was shown that paraffin immunohistochemistry was useful for the practical histopathological diagnosis of NHML even in the area where human T-cell leukemia virus type 1 is endemic.

Effects of prolonged treatment with beta-adrenoceptor antagonist, carteolol on systemic and regional hemodynamics in stroke-prone spontaneously hypertensive rats.

The present study was designed to determine the regional hemodynamic effects of prolonged beta-adrenergic receptor inhibition in conscious stroke-prone spontaneously hypertensive rats (SHRSP) using a radioactive microsphere method. When the regional blood flow was compared between 10 and 30 weeks of age, the age-related changes in organ blood flow were observed in several organs, i.e., the reduction of flow rate in kidney, adrenal gland and intestines. The reduction of flow rate in these organs contributes strongly to the age-related rise of total peripheral resistance. Carteolol, a beta-adrenoceptor antagonist, was given at a dose of 10 mg/kg/d from 10 to 30 weeks of age. These animals gained more weight than the untreated control SHRSPs, and heart rate was reduced significantly. Blood pressure was not affected. However, the prolonged treatment with carteolol prevented the age-related reduction of the blood flow rate in the kidney, adrenal gland and intestines. Thus, our findings indicate that carteolol had appreciable and beneficial effects on the maintenance of flow rates in the above organs of SHRSP without any change in blood pressure.