Immunohistochemical characterization of undifferentiated carcinomas of the ovary

Immunohistochemical characteristics of undifferentiated carcinomas of the ovary were examined using formalin-fixed, paraffin-embedded tissues with an avidinbiotin staining approach. Eight cases were collected from the pathology files of our Institute from a total of 214 recorded malignant ovarian tumors. For immunostaining, antibodies reacting with epithelial membrane antigen (EMA), pankeratin, vimentin, CA 125, CA 19-9, carcinoembryonic antigen (CEA), -fetoprotein (AFP), -l-antitrypsin (AT), epidermal growth factor receptor (EGFR), c-erbB-2,bcl-2 and p53 proteins were used. All the cases examined were positive for EMA and pankeratin, specific markers for epithelial tumors, negative for the non-epithelial tumor marker, vimentin, and also positive for EGFR. Interestingly, only one case was positive for CA 125, despite it being one of the commonest reported indicators of ovarian cancer. CA 19-9 was positive in 7 cases, CEA in 5, AFP in 2, AT in 6 and c-erbB-2 protein in 4. Two cases were positive for p53 protein, and in 1 of these positive staining forbcl-2 was also observed. These results indicate that the epithelial nature is well preserved in undifferentiated ovarian carcinomas, although consistently positive reactions were not observed within the cases for some antigens. They further celarly show that a negative signal for CA 125 can not be considered to exclude the possibility of a primary ovarian tumor.Abbreviations EMA epithelial membrane antigen - EGFR epidermal growth factor receptor     

Osteoclastic giant cell tumour of the pancreas

Osteoclast giant cell tumours are bone tumours that occur in adults, and that are considered benign by WHO but locally aggressive. Strictly identical tumours are described in the pancreas, without simultaneous bone localization. We report the case of a 62-year woman with an osteoclast giant cell tumour of the distal pancreas, without any epithelial component, which was diagnosed after pancreatic resection and with no signs of recurrence after a 24-month follow-up. These pancreatic tumours are rare, with a very poor prognosis, an unclear histogenesis; they are often confused with pleomorphic or undifferentiated pancreatic carcinomas including a component of osteoclast giant cell. These osteoclast giant cell tumours of the pancreas usually present as large cystic tumours. In certain cases, complete resection can result in long-term survival.     

Inflammatory fibroid polyp: an immunohistochemical study

BACKGROUND: Inflammatory fibroid polyp is a localized lesion, which arises in the submucosa of the gastrointestinal tract, most often in the stomach.Although it is generally believed to represent a reactive, nonneoplastic condition, its histogenesis remains controversial. AIM: To study inflammatory fibroid polyp by immunohistochemistry in an attempt to further clarify their histogenesis. MATERIAL AND METHODS: Nine cases were studied by immunohistochemistry using a panel of antibodies against smooth-muscle actin, vimentin, S-100 protein, factor VIII- R.Ag and macrophage (HAM-56). RESULTS: There was a strong diffuse positive staining pattern in the spindle cells with vimentin antibody. A patchy staining for smooth-muscle actin was observed in these cells. Immunophenotyping revealed a heterogeneous reaction with HAM-56. In edematous areas, HAM-56-positive cells show voluminous cytoplasm and reniform nuclei. In cell-rich areas, the HAM-56-positive cells had fusiform cytoplasm. Stains for S-100 and factor VIII RAg were negative in the proliferating elements. CONCLUSIONS: The present immunohistochemical study refutes the suggested neural or vascular nature of the lesion. The strong positivity for vimentin in all cases suggests a major component of spindle cells best recognizable as fibroblasts. These results would favor the existence of a span of morphological and immunohistochemical patterns possibly indicating evolutive phases of an inflammatory reaction.     

Undifferentiated carcinoma with osteoclast-like giant cells of the pancreas]

Undifferentiated carcinoma with osteoclast-like giant cells is a rare neoplasm of exocrine pancreas. Till recently, some cases have been reported, however histogenesis of the tumors are controversial and their characteristic findings have not been described yet. Thirty five-year-old men and 75-year-old men were presented with upper abdominal pain and a palpable mass. On computed tomography, one case showed a well enhancing solid tumor with low density and the other was showed a mainly cystic tumor with peripheral enhancement in the body and tail of the pancreas. One case accompanied multiple metastatic liver masses with subhepatic lymph node enlargement. Tumor staining was seen on angiography. Biopsy and pancreatectomy were performed. Pathological findings revealed tumors composed of neoplastic spindle shaped or pleomorphic large cells with scattered non-neoplastic osteoclast-like giant cells. In one case, there were small foci of adenocarcinoma components in the periphery of the tumor. On immunohistochemical stain, neoplastic cells showed focal positivity for epithelial membrane antigen and vimentin. Tumors were diagnosed as undifferentiated carcinoma with osteoclast-like giant cells. We report these rare cases with a review of literature.     

Expression of KIT (CD117) in Biphasic Pulmonary Blastoma. Novel Data on Histogenesis

Biphasic pulmonary blastoma (BPB) is a rare primary neoplasm of the lung and its histogenesis is still uncertain. It has been proposed that BPB is derived from mesoderm or endoderm. Others suggested an origin from a single pluripotential cell. We present a case of a BPB with emphasis on expression of the stem cell factor receptor KIT (CD117). We describe a 61-year-old male patient with a BPB of the upper right lobe. Immunohistochemical analysis was performed using a panel of several antibodies including anti-CD117. Strong cytoplasmic expression of CD117 was found in the epithelium (cytokeratin-positive) as well as in the spindle cells (cytokeratin-negative). Expression of CD117 in both mesenchymal and epithelial cells suggests a single origin and supports the idea that BPB arises from a pluripotential cell that can differentiate into both stromal and epithelial morphologies. The role of CD117 in the pathogenesis of BPB and its possible therapeutic relevance require further investigation.     

Concurrent occurrence of gastric adenocarcinoma and duodenal neuroendocrine cell carcinoma: a composite tumour or collision tumours ?

BACKGROUND: Neuroendocrine cell (NEC) carcinoma is occasionally accompanied by adenocarcinoma but the relationship between these two morphologically distinct tumours is unclear. Two hypotheses have arisen regarding the mechanism for the association of adenocarcinoma and NEC carcinoma. One is that both are derived from a common multipotential epithelial stem cell. The second hypothesis is that adenocarcinoma and NEC carcinoma arise from a multipotential epithelial stem cell and a primitive NEC, respectively. AIMS: To elucidate the relationship between the two histologically distinct tumours, adenocarcinoma of the stomach and NEC carcinoma of the duodenum. PATIENT/METHODS: We present a case in which the tumour extended across the pyloric ring, the gastric portion of which revealed adenocarcinoma while the duodenal portion showed argyrophil NEC carcinoma. The two histologically distinct lesions of the tumour were examined by immunohistochemistry and genetic analysis of p53. RESULTS: The gastric region was negative for chromogranin A staining but positive for carcinoembryonic antigen (CEA) staining. In contrast, the duodenal region was positive for chromogranin A but negative for CEA. All tumour regions showed a point mutation in p53 gene at exon 7 (GGC (glycine)-->GTC (valine) at codon 245). The distal portion of the duodenal tumour showed an additional point mutation in p53 gene at exon 5 (GCC (alanine)-->GTC (valine) at codon 129). CONCLUSIONS: The two histologically distinct tumours, adenocarcinoma of the stomach and NEC carcinoma of the duodenum, appear to be derived from a common epithelial cell.     

Tissue polypeptide antigen expression in human prostate tumors

Summary Thirty-seven specimens of benign and malignant prostatic tumors were studied for the localization of tissue polypeptide antigen (TPA) by an avidinbiotin-peroxidase complex technique. In addition, 23 metastases of prostatic carcinoma in other organs and 12 nonepithelial tumors of prostate also were studied. All benign and malignant tumors of epithelial origin, including their metastasis, stained positively. Non-epithelial tumors were uniformly negative. In the metastatic lesions, small foci of tumor cells and even single tumor cells could be identified by TPA staining. Immunohistochemical localization of TPA appeared to be a useful tool for assessing the micrometastases of prostatic carcinoma in other organs, especially lymph nodes, or elucidating the epithelial origin of an otherwise undifferentiated prostatic cancer.This work was supported in part by the Adult Disease Memorial Institute, Tokyo, Japan     

Surface marker analysis in acute myeloid leukaemia and correlation with FAB classification

The immunological phenotype of blast cells in 102 patients with acute myeloid leukaemia (AML) was analysed with a panel of 20 monoclonal antibodies and the enzyme terminal transferase, and correlated with the FAB classification. Although a partial correlation between these two approaches could be observed, almost every morphological group contained patients from more than one immunological phenotype. The M1 and M5a leukaemias showed the most undifferentiated phenotype, often lacking in specific myelomonocytic antigens. The M3 formed a uniform group defined as My7+, Ia-, FMC8+, a phenotype which was also observed in two cases of the microgranular variant. The granulocytic (CDw15) and monocytic (CDw14) antibodies crossreacted with some M5b and M2 leukaemias, respectively. Compared with M5a, the M5b leukaemias showed a large increase in the expression of CDw14 antigen, confirming the validity of the morphological differentiation. Glycophorin-A was present in four out of five M6 leukaemias. TdT activity was demonstrated in 10% of AML cases, with a higher incidence among the monocytic variants: M4 and M5-. Eleven AML were considered as unclassifiable according to the FAB criteria and in seven of them a megakaryoblastic cell population (GP IIb/IIIa+, GPIb+) was demonstrated; this confirms the need to include the subgroup of megakaryoblastic leukaemias within the AML. Finally, a possible immunological classification for AML is proposed.     

Immunoscintigraphy with emission tomography in cancers of the thyroid

Immunoscintigraphy (IS) consists of in vivo body structure imaging using a specific labelled antibody to an antigen concentrated in the structure under study. Technically, the image contrast is better when IS is performed with a computerized emission tomography system. High concentrations of carcinoembryonic antigen (CEA) have been reported in medullary thyroid carcinoma and thyroglobulin (Tg) is a marker for differentiated thyroid carcinoma. We used injections of 131-I labelled monoclonal antibodies to CEA and Tg to detect thyroid tumours. A feasibility trial using anti-CEA antibodies gave very encouraging results. However, only tumours larger than 10 cm3 could be detected. Contradictory results were obtained using anti-Tg antibodies but this data must be considered as preliminary. Various means of improving the method and the concept of accessibility of the antigen to the antibody in vivo are discussed. This study shows IS to be a promising experimental technique. Further studies are required to define its clinical indications before it can be advocated for routine use.     

Localization of factor VIII-procoagulant antigen: an immunohistological survey of the human body using monoclonal antibodies

Various organs, including liver, spleen, heart, lung, kidney, intestines, lymph nodes, pancreas, bone marrow, and thymus, were investigated for the presence of factor VIII-procoagulant antigen (VIIICAg) and factor VIII-related antigen (VIIIRAg), using a panel of monoclonal antibodies directed to factor VIII-von Willebrand factor in combination with a sensitive immunoperoxidase staining technique. In addition to hepatic sinusoidal endothelial cells, the presence of VIIICAg was demonstrated in mononuclear cells sporadically present in lymph nodes, in the alveolar septa of lung, and in the red pulp of spleen. The identity of these mononuclear cells could not be unequivocally determined. Based on morphological criteria, however, it is tentatively concluded that these cells are nonlymphoid and belong to the mononuclear phagocyte system. The presence of VIII-RAg was confined to vascular endothelial cells, hepatic sinusoidal endothelial cells, cells lining the venous sinuses of the red pulp of the spleen, cells lining renal glomeruli and lung capillaries, platelets, and megakaryocytes.     

Immunohistochemical study of carbohydrate antigen expression in gastric carcinoma

The expression of carbohydrate antigens in malignant and non-malignant gastric mucosa was studied immunohistochemically using the monoclonal antibody AH6 directed to Ley antigen, FH2 directed to Lex antigen, and FH6 directed to sialyl-Lex antigen. Formalin-fixed gastric tissue resected from 54 patients with gastric cancer and 20 patients with gastric ulcer were tested. The incidence of positive cases in gastric cancer patients with each antibody was as follows: AH6;85%, FH2;74%, FH6;74%. The Lex antigen was expressed in 81.5% of cases histologically classified as undifferentiated type, and 66.7% of cases classified as differentiated type. It was expressed in a higher incidence in early stage cancer (93.3%) than in advanced stage cancer (66.7%). Sialyl-Lex antigen was detected in more cases of differentiated type (88.9%) than in those of undifferentiated type (59.3%), whereas none of 8 early cancers of undifferentiated type expressed the antigen. The incidence of the expression of Ley antigen did not differ in relation to histological type or invasiveness. Lex and Ley antigens were detected in noncancerous gastric epithelium. Sialyl-Lex antigen was not detected in the normal fundic gland region. These results demonstrate that Lex antigen may be a differentiation-associated antigen, and sialyl-Lex antigen might be useful as a marker of differentiated cancer and an indication for invasion of undifferentiated cancer.     

Expression of Ley antigen as a phenotypic marker of apoptosis in alcoholic hepatitis

Apoptosis is a type of cell death which is clearly distinguishable from necrosis in its morphological and biochemical features. To clarify the role of apoptosis in alcoholic liver injury, we investigated the expression of apoptosis-related Le^y antigen by immunohistochemistry in liver samples from patients who suffered from alcoholic liver diseases. Liver biopsy samples were taken from 20 patients who drank more than 80 g ethanol per day on average. Indirect immunohistochemical staining was carried out using anti-cytokeratin (CAM 5.2) and anti-Le^y (BM-1/JIMRO) antibodies. To examine the relationship between Mallory bodies and apoptosis, double staining was performed using both antibodies. In alcoholic hepatitis, many Mallory bodies were stained with anti-cytokeratin antibody in hepatocytes of the centrilobular area. Le^y antigen was also detected in hepatocytes in the same area. Immunohistochemical double staining showed that some of the hepatocytes containing Mallory bodies were stained with anti-Le^y antibody. Few hepatocytes expressing Le^y antigens, however, were observed in other types of alcoholic liver diseases including steatosis, fibrosis and cirrhosis. From these results, it is concluded that apoptosis is also involved in alcoholic hepatitis and that hepatocytes containing Mallory bodies may be eliminated by apoptosis.     

The value of bone marrow examination for tumor staging in breast cancer

Summary The purpose of our study was to investigate the value of cytokeratin antibodies for identifying bone marrow involvement in breast cancer patients who showed no evidence of distant metastases using noninvasive tumor staging procedures. Bone marrow for histological (biopsy) and immunocytochemical (aspiration) evaluation was obtained from the anterior iliac crest from 50 unselected consecutive women during surgical treatment of the primary tumor. The histological examination was done on nondecalcifed bone sections. The immunocytochemical studies were carried out on interface smears of the bone marrow aspirates. For staining, cytokeratin antibodies (PKK 1) and the immune alkaline phosphatase method was used. Cytokeratin-positive cells were found in 4 of the 50 cases (8%). Of those 4 patients, however, 2 also showed evidence of neoplastic bone marrow infiltration histologically. We thus were able to prove that immunocytochemistry on aspirates is superior to conventional histology in identifying tumor in bone marrow. Nonetheless, our results clearly fell below the rate found in previous studies where epithelial membrane antigen antibodies were used.     

A diagnostic approach to primary malignant lymphoma of the intestine

Thirty-one cases of primary malignant lymphoma of the intestine diagnosed since 1980 were studied. Being reviewed the clinical data all cases were reclassified histologically according to Isaacson's new classification in 1988. The immunohistochemical staining were performed with epithelial, B-cell, T-cell and histiocytic markers for early diagnosis of the disease. The results showed male, youth and laborer predominance with a short courses and poor prognosis, Colonoscopy and X-Ray found "malignant lesions" in characteristics. Histologically, High-grade and B-cell lymphoma were predominance. MALT lymphoma and the combination of low to high-grade lesions were found. Immunohistochemical staining is quite helpful in the differentiation of carcinoma, malignant histiocytosis. Finally, the procedure of early diagnosis of the disease was proposed.     

Cell surface antigens of human ovarian and endometrial carcinoma defined by mouse monoclonal antibodies.

Mouse monoclonal antibodies to several cell surface antigens of human ovarian and endometrial carcinomas have been produced. The distribution of the antigens was determined by mixed hemagglutination assays on 153 normal and malignant cell cultures of various types and by immuno-peroxidase staining of frozen sections of 27 normal adult and 24 fetal tissues. Five distinct antigens were characterized. MD144 antigen was detected on only a single ovarian carcinoma cell line and has the biochemical properties of a lipid. MH55 antigen is weakly expressed on ovarian and uterine cancer cell lines but not on other cells and tissues tested. MF61 antigen was detected on an ovarian carcinoma and some renal carcinoma cell lines but not on other cell lines tested. It was also detected by immunoperoxidase staining in the noncellular follicles of the thyroid and in uterine glandular epithelial cells. This antigen also has the properties of a lipid. MF116 antigen was detected on a proportion of ovarian, uterine, renal, and bladder carcinoma and neuroblastoma cell lines and on normal kidney epithelial cell cultures but not on other cell lines tested. It was not detected in sections of any normal tissue tested using the immunoperoxidase method. MF116 was readily detected in the spent culture medium but not in detergent-solubilized extracts of metabolically radiolabeled cells. This shed antigen is a glycoprotein of Mr 105,000 and isoelectric point lower than pH 4.0. MH94 antigen was detected on a proportion of ovarian, uterine, colon, breast, lung, cervical, and pancreatic carcinoma cell lines. In tissue sections it was detected in many but not all epithelia, predominantly in secretory epithelial cells. Antibody MH94 did not immunoprecipitate a detectable antigen.     

Medullary thyroid carcinomas express chromogranin A and a novel neuroendocrine protein recognized by monoclonal antibody HISL-19

PURPOSE AND METHODS: A number of endocrine peptides and proteins are expressed by medullary thyroid carcinoma (MTC). The expression of two newly appreciated neuroendocrine tumor markers, chromogranin A (CgA) and the endocrine antigen defined by monoclonal antibody HISL-19, was determined in 14 MTCs by immunohistology to evaluate the clinical utility of these markers in the diagnosis of MTC. Papillary, follicular, and undifferentiated thyroid tumors were also evaluated along with an MTC cell line. The same tissues were evaluated with antibodies to human calcitonin. RESULTS: All human calcitonin antibodies were found to react with the MTCs. In addition, all MTCs were reactive for CgA and the antigen detected by antibody HISL-19. CgA was generally present in the human calcitonin-containing cells, whereas the HISL-19 antigen had a more distinctive distribution. The other thyroid tumors failed to show reactivity with any of the three antibodies. CONCLUSION: Our results demonstrate that, in addition to human calcitonin, MTCs commonly express CgA and the antigen defined by antibody HISL-19. Our observations thus add to the repertoire of endocrine substances produced by MTC. These studies also demonstrate the clinical value of immunohistologic procedures for two novel antigens in distinguishing MTCs from other thyroid tumors.     

Surface markers in acute non-lymphoid leukemia: Analysis with a panel of 36 monoclonal antibodies

Summary The reactivity of a panel of monoclonal antibodies was studied in fifty-four cases of acute myeloid (AML) or undifferentiated (AUL) leukemias. Thirty-six antibodies from the Myeloid section of the Second Workshop on Human Leukocyte Differentiation Antigens were used in an indirect immunofluorescence assay. The antibodies could be classified into three groups recognizing respectively granulocytic, monocytic or granulomonocytic leukemias. Most antibodies stained erythroblastic and megakaryoblastic leukemias. In each group, it was possible to define antibodies staining either the less differentiated forms (FAB M 1 and M 5 a) or the more differentiated forms (M 2, M 3, M 4 and M 5 b). Six out of eight AUL were stained by some of the antibodies (mainly from the monocytic group). However, a heterogeneity of stainings in a same blast population was observed.     

Biphasic sarcomatoid carcinoma of the thyroid: an exceptional localization of a rare tumor

Thyroid sarcomatoid carcinoma is a rare and aggressive neoplasm composed of a follicular carcinoma which is contiguous or admixed with a pleomorphic spindle cell component. We report the case of a thyroid tumor reputed to have a poor outcome, in a 62-year-old woman. The radical thyroidectomy specimen was totally invaded. Results of immunoperoxidase staining for thyroglobulin and epithelial markers were positive in the areas of follicular carcinoma and negative in the sarcomatous component. The patient died a few days later due to septic shock. The epithelial and mesenchymal components of thyroid carcinosarcoma were both part of the neoplastic parenchyma and evolved from a single common stem cell, in agreement with the hypothesis that the tumors are of monoclonal origin. The definition of this tumor as its histogenesis and prognostic are discussed.     

The production and characterization of monoclonal antibodies to the human thyroid microsome

By suitable immunization of mice and fusion of their spleen cells with a non-secretor mouse myeloma line, monoclonal antibodies have been produced which react with the human thyroid microsomal (M) antigen. These monoclonal antibodies showed no reactivity by enzyme-linked immunoassay with liver microsomes or thyroglobulin and their specificity was confirmed by immunolocalization studies, in which they showed the staining characteristics of human M antibodies. All four monoclonal antibodies tested were immunoglobulin M; three were cytotoxic to thyroid cell monolayers. The lack of cytotoxicity with the fourth monoclonal supports the concept that certain epitopes of the M antigen may be partially or completely absent at the thyroid cell surface. These monoclonal antibodies should permit further characterization of the thyroid M antigen in view of their absence of cross-reactivity with thyroglobulin.