Immunohistochemical characterisation of pulmonary hyaline membrane in various types of interstitial pneumonia

AIMS: Hyaline membrane (HM) in diffuse alveolar damage (DAD) pattern is frequently detected in the acute stage of interstitial pneumonia (IP). To determine the exact nature of HM, we investigated immunohistochemically 25 cases of HM-containing IP. METHODS: The cases examined using various kinds of antibodies were four cases associated with rheumatoid arthritis, five with usual interstitial pneumonia, two with dermatomyositis, five with viral infection, one case with progressive systemic sclerosis and eight cases caused by other agents. RESULTS: HM mostly reacted with antibodies to PE10 (SP-A), Factor VIII, KL-6 and EMA and, interestingly, stained for AE1/AE3, CK19, and Hup-1 in some cases, but was negative for PTAH staining. However, the immunoreactivities of HM varied even within the same disease or section. CONCLUSIONS: The immunohistochemical heterogeneity of HM suggests that HM may be formed by different mechanisms in various types of IP. Our findings also suggest that the main components of HM are derived from alveolar epithelial cells and their proteins, some including cytoplasmic element of CK19, and also from serum factors, but not fibrin. The immunohistochemical characteristics of HM in DAD pattern will aid understanding of the significance of HM formation in IP.     

Extracellular collagenous spherules in salivary gland tumors. Immunohistochemical analysis of laminin and various types of collagen.

Collagenous spherulosis is a benign breast lesion involving lobular acini and ductules and containing eosinophilic spherules measuring up to 100 microns in diameter. We present an immunohistochemical analysis of similar collagen-rich spherules that are also found in salivary gland tumors. These collagenous spherules contain varying amounts of acidic mucins, elastin, basement membrane proteins including type IV collagen and laminin, and considerable amounts of interstitial collagen types I and III. Types II and VI collagen were not detected in collagenous spherules of salivary gland tumors. The cells surrounding these collagenous spherules expressed muscle actin, S100 protein, vimentin, and cytokeratins 8, 18, and 19, indicating that these cells have myoepithelial characteristics.     

Immunohistochemical study of fiber types in human extraocular muscles

Fiber types in human extraocular muscle (h-EOM) were examined immunohistochemically with antibodies against slow tonic (anti-ALD) and slow twitch (anti-SOL) myosins. Four types of muscle fiber in h-EOM were distinguishable according to their reactivities with these antibodies. Groups 1 and 2 fibers reacted with both antibodies, group 1 fibers showing stronger reactivity than group 2 fibers with anti-ALD. Group 3 fibers reacted only with anti-SOL. Group 4 fibers did not react with either antibody. The latter were the most common, and were the main fibers in both the peripheral (outer orbital) and central zones of h-EOM. The next most common were group 1 fibers, which were located mainly in the peripheral layer. Group 2 fibers were less common, but were the second most common type in the central layer. Group 3 fibers were only minor constituents. Multiple innervations were observed in some fibers of groups 1 and 2, and group 1 fibers were suggested to be slow tonic myofibers in h-EOM. These specific immunohistochemical and physiological features of h-EOM seem to be the basis of the low morbidity seen in the usual types of muscular dystrophy.     

Immunohistochemical study of rat soleus muscle in various modes of denervation

Immunohistochemical study with monoclonal antibodies against fast myosin heavy chains showed that excision of a fragment of sciatic nerve does not change, while compression of the nerve decreased the relative content of fast muscle fibers in rat soleus muscle. The content of fast fibers in the muscles of contralateral limbs decreased in both models of denervation. Hence, contralateral limbs cannot be used as the control in experiments with disturbed neurotrophic function. Possible mechanisms underlying changes in the immunohistochemical profiles of experimental and contralateral limbs are discussed.     

Immunohistochemical study of arginase 1 and 2 in various tissues of rats

Arginase 1 and arginase 2 catalyze the hydrolysis of arginine to ornithine and urea. The localization of these enzymes was studied in various tissues in Sprague-Dawley rats by immunohistochemistry and Western blotting. Western blot analysis showed that both arginase 1 and 2 were differentially expressed in the various organs examined. Arginase 1 was expressed at high levels in the liver, at moderate levels in the pancreas, and at low levels in the cerebrum, cerebellum, spinal cord, stomach, small and large intestines, kidneys, lungs, and spleen. The levels of arginase 2 immunoreactivity were high in the kidneys and pancreas, and moderate in the cerebrum, spinal cord, stomach, small intestine, large intestine, and lungs; the levels were very low in the liver and spleen compared with that in the cerebellum. Immunohistochemical analysis largely confirmed the results of the Western blot analysis. These findings indicate that the levels of arginase 1 and 2 varied among organs, suggesting that the arginase isoforms may play organ-specific roles in the urea cycle.     

Immunohistochemical profile of ephrin A4 expression in human osteosarcoma

Ephrin receptors and ephrin ligands constitute one of the largest groups of tyrosine kinases. The division of ephrin receptors into type A or type B is determined by their ligand‐binding specificities. Ephrin A4 as a ligand has a broad capacity to bind and stimulate different subtypes of ephrin A receptors. Little is known about the role of ephrins generally and ephrin A4 particularly in osteosarcoma. Ephrin A4 was immunohistochemically assessed on archival material from 46 primary osteosarcoma cases, 10 metastatic pulmonary lesions and 20 normal control bone specimens. Ephrin A4 was expressed in 100% of normal bone specimens, in 84.4% of primary osteosarcoma cases and in all metastatic pulmonary lesions. Cytoplasmic and nucleocytoplasmic patterns of ephrin A4 immunoreactivity were observed, with the predominance of the latter pattern in normal bone (100%), and in 43.5% of primary osteosarcoma cases, which showed a higher intensity of expression compared with normal bone (p<0.05). The cytoplasmic pattern is the only staining pattern seen in metastatic cases, which may suggest its role in enhancement of invasion and metastasis. The differences in the distribution of the two patterns of ephrin A4 may indicate a different biological activity of this molecule depending on its localization. The nuclear localization of ephrin A4 requires further investigation to clarify the mechanism and the significance of the nuclear trafficking of ephrin A4.     

Skeletal muscle in polymyositis. Immunohistochemical study.

Thirty-two patients with adult-onset polymyositis uncomplicated by cancer or systemic connective tissue disease were studied. Muscle biopsy specimens were examined with direct immunofluorescence microscopy and results were compared with those in 94 control subjects. Sarcolemmal and sarcoplasmic staining were observed in both groups and considered to be nonspecific. Immune deposits in the muscle microvasculature were present in some cases of systemic lupus erythematosus and dermatomyositis but were not present in polymyositis. Our data suggest that the finding of vascular immunofluorescence excludes the diagnosis of adult polymyositis and implies that the pathogenesis of this disease and other idiopathic inflammatory myopathies may differ.     

Immunohistochemical study of lichen planus.

The distribution of IgM and IgG in 20 cases of lichen planus and five cases of non-specific inflammation were studied by the unlabelled antibody-enzyme (PAP) technique. Routine paraffin sections were used. In lichen planus deposits of immunoglobulins were seen in and around epithelial cells, in colloid bodies, at epidermo-dermal junctions, and in some inflammatory cells. All cases examined for IgM and eight out of 13 cases examined for IgG were positive. The peripheral migrating epidermal cells were mostly negative. The application of the PAP technique to skin biopsy and the significance of the findings in lichen planus are discussed.     

Immunohistochemical analysis of p27 (Kip1) in human pituitary glands and in various types of pituitary adenomas

p27 (Kip1) plays regulatory roles in the cell cycle by inhibiting the activity of cyclin dependent kinases (CDKs). This immunohistochemical study is aimed at elucidating the expression of p27 in human pituitary and in various types of pituitary adenomas in order to clarify its role in the regulation of proliferation. Sixteen normal pituitary glands and 179 human pituitary adenomas were used for immunohistochemical studies. The tissues were fixed in 10% formalin and embedded in paraffin. Indirect peroxidase method was performed after heat-induced antigen retrieval using a monoclonal antibody against p27 protein. p27 protein was expressed in the nuclei of all 16 normal human pituitary glands. p27 protein was also expressed in 128 of 179 cases of pituitary adenomas (71.5%). A marked decrease of p27 expression was noted in ACTH-secreting adenomas, 8/20 (40.0%), compared with other types of pituitary adenomas—GH-secreting adenomas, 35/46 (76.1%); PRL-secreting adenomas, 22/33 (66.7%); TSH-secreting adenomas, 8/11 (72.7%); and nonfunctioning adenomas, 55/69 (79.7%). These results suggest that p27 may play some role in the regulation of proliferation in all types of pituitary adenomas. The lower levels of p27 in ACTH-secreting adenoma is of particular interest with respect to the intermediate lobe-derived pituitary tumor developed in p27 knockout mice.     

Expression of osteopontin and osteonectin in breast cancer.

Osteopontin (OP) and osteonectin (ON) are bone matrix proteins produced by mammary and other cancers. These proteins may play a role in tumor invasion and metastasis through integrin-mediated signal transduction. We evaluated expressions of OP and ON in 253 resected infiltrating ductal carcinoma of the breast, using immunohistochemical staining and follow-up data. OP and ON were detected 87.4% and 54.2% of 253 cases, respectively. The OP and ON positive staining were localized in the cytoplasm of carcinoma cells. OP and ON did not correlate with various clinicopathological parameters, such as age, lymph node involvement, tumor size, histologic grade, expression of p53 and estrogen receptor (ER). In the multivariate model, lymph node involvement and histologic grade were statistically significant prognostic factors. Assessed by a log rank test, the 5-year-survival rates of OP and ON positive groups and their negative groups were not statistically different. In conclusion, OP and ON immunopositivity of infiltrating ductal carcinomas of the breast provide no additional prognostic information in this study.     

骨肉瘤的病理类型及其病理特点

骨肉瘤的病理类型及其病理特点丘钜世朱全胜王连唐骨肉瘤是发生于间叶组织的具有直接形成骨质或骨样组织的恶性肿瘤，是恶性骨肿瘤中最常见者。骨肉瘤多为原发性，但也可在原先某种骨疾患（如骨母细胞瘤、骨纤维结构不良、骨软骨瘤、骨巨细胞瘤及骨的Ｐａｇｅｔ病等）的基...     

Immunohistochemical characterization of ovarian borderline tumors of intestinal and mullerian types.

A panel of monoclonal antibodies (MAbs) including MOv2, MOv8, anti-CA 19-9, and anti-carcinoembryonic antigen (CEA), and a polyclonal antibody against CEA, was applied to three types of ovarian borderline tumors. The tumors included 19 intestinal-type mucinous borderline tumors (IMBTs), 22 endocervical-like mucinous borderline tumors (EMBTs), and 23 mixed-epithelial borderline tumors (MEBTs); the latter two tumors are of mullerian type. Statistically significant differences in the percentage of positive IMBTs compared to the mullerian tumors were seen for anti-CEA and MOv8; strikingly different staining patterns were also seen. IMBTs were more often and more diffusely CEA-positive than were the mullerian tumors; within the mullerian tumors, only one type of cell, the indifferent eosinophilic cell, was consistently CEA positive. The MAbs MOv2, MOv8, and anti-CA 19-9 showed more extensive positivity in the mullerian tumors than in the IMBTs. This study highlights the differences in cell types between IMBTs and these two types of mullerian borderline tumors.     

Immunohistochemical study of 158 lung carcinomas.

Lung carcinomas were studied immunohistochemically and the results were related to type of tissue sample (bronchoscopic biopsies, surgical specimens, autopsies). All cytokeratins (CAM 5.2, PKK-1, AE1/AE3) reacted with virtually all adenocarcinomas, most squamous, and 65% of the large cell carcinomas, while CAM 5.2 was most efficient with the small cell carcinomas. CEA stained 33% and 60% of the small and large cell carcinomas, respectively, most adenocarcinomas, and 84% of the squamous cell carcinomas, among which staining decreased with dedifferentiation and was often focal. EMA reacted with 90%, and NSE with 20% of all histological types. There was no staining for NF. All antibodies, except EMA, were more efficient with surgical specimens. Our study implies that the cytokeratins we used work better with surgical material, but are generally comparable to monospecific cytokeratin antibodies. Also, EMA is a reliable marker for epithelial differentiation with all types of tissue samples. Moreover, CEA negativity in several poorly differentiated lung carcinomas might have implications in the differential diagnosis against pleural mesothelioma.     

A case study of Woringer-Kolopp disease. Immunohistochemical study

Woringer-Kolopp disease is a localized epidermotropic T cell lymphoma with good prognosis. We describe a 79-year-old man with an erythematous scaly plaque of the foot. Clinical diagnosis of psoriasis, parapsoriasis and fungal infection were proposed. Histopathological and immunohistochemical findings were characteristic of Woringer-Kolopp disease. We observed an immunohistochemical positivity of tumour cells for the anti-CD103 antibody (alphaEB7 integrin) according to the epidermotropism of this localized cutaneous T cell lymphoma.     

Immunohistochemical evaluation of keratin 20 expression in intestinal metaplasia types I to III.

AIMS: To investigate differences in expression of keratin 20, a cytoskeletal protein in gastrointestinal epithelial cells, in completely differentiated intestinal metaplasia (type I) and incomplete metaplasia (types II and III). METHODS: Gastric biopsy specimens from 66 patients with intestinal metaplasia were analysed immunohistochemically. Expression of keratin 20 was quantified as the percentage of immunoreactive cells on the tips, the upper, and deep foveolae. RESULTS: In all specimens keratin 20 was found on the tips and in the upper foveolae of intestinal metaplasia. Keratin 20 was not observed in the deep foveolae. No differences were seen between the antrum and the body. Expression patterns were comparable between types I and III. In type II, however, lower immunoreactivity was found. Both the differences between types I and II as well as between types II and III were significant (p < 0.05). CONCLUSIONS: Keratin 20 is expressed in metaplastic mucosa as a result of intestinal differentiation. Positive staining found exclusively in juxtaluminal cells occurs only in mature cells containing keratin 20. Lowered immunoreactivity in type II compared with types I and III indicates the different nature of type II intestinal metaplasia. Further studies are needed to shed light on the basic fundamental mechanism responsible for this.     

Immunohistochemical study of fibronectin in experimental myocardial infarction.

Light microscopic immunohistochemical studies were performed to evaluate the distribution of fibronectin in paraffin sections of p-formaldehyde-fixed normal rat hearts and the hearts of rats that had undergone ligation of the left coronary artery. A peroxidase-labeled antibody technique was used, together with appropriate immunohistochemical control procedures, for the localization of fibronectin in normal hearts and in the hearts of sham-operated animals. Fibronectin was localized in the interstitial space between myocytes, and beneath arterial, venous, and capillary endothelium. At 4 hours after coronary ligation, fibronectin was localized in a patchy fashion in the cytoplasm and interstitial space of some of the myocytes in the area supplied by the ligated vessel. At 24 hours, there was more intense, homogeneous staining in necrotic myocytes in the infarcted area and in the capillary endothelium in the border zone. At 48 hours, the intensity of staining for fibronectin was maximal in and between the necrotic myocytes in the center of the infarct and in proliferating and migrating capillaries and fibroblasts in the border zone. Similar patterns of localization were observed at 3 and 7 days after coronary ligation, but with progressive decreases in the intensity of staining. Two sources of fibronectin appeared to have contributed to these changes: plasma fibronectin diffusing through damaged blood vessels would account for the early staining observed in necrotic myocytes in the center of the infarct, whereas de novo synthesis of fibronectin by connective tissue cells and endothelial cells in sprouting capillaries would be responsible for the subsequent staining observed in viable capillaries in the border zone of the infarct. Known properties of fibronectin in vitro, combined with these in vivo observations, indicate that fibronectin may influence the thrombotic, inflammatory, angiogenic, and fibrotic processes involved in infarct healing.     

Immunohistochemical study of intimal microvessels in coronary atherosclerosis.

Two hundred ninety-nine human coronary artery paraffin-embedded tissue blocks were examined for intimal microvessel invasion by probing for factor VIII-associated antigen with indirect immunofluorescence and high resolution confocal microscopy. The results obtained confirm that intimal microvessels originate in the adventitia and show that the richness of intimal microvessels is strongly positively correlated with intimal thickness and negatively correlated with relative lumen size. A number of plasma constituents were examined in serial sections. Comparison of immunofluorescence distribution patterns of these components with intimal microvessel distribution patterns reveals that intimal microvessels leak plasma albumin into artery walls, exude fibrinogen, and are associated with the build-up of plasma cells within atherosclerotic lesions. Therefore, intimal microvessels are demonstrated to play important roles in the development of atherosclerosis.