Epithelial Membrane Antigen Immunohistochemistry Research Articles

English

BACKGROUND Inconclusive cytomorphology often results due to failure to distinguish between adenocarcinoma cells from benign, reactive, atypical mesothelial cells in effusion specimens. To resolve such dilemmas, auxiliary techniques like immunohistochemistry were utilised to reach a definitive diagnosis for better treatment and management of patients. We wanted to compare cytodiagnosis achieved on cell block preparations with the cytodiagnosis on conventional smear and perform immunohistochemistry for epithelial membrane antigen (EMA), calretinin, desmin, vimentin and E-cadherin on cell block preparation of the fluids in cases of indistinguishable cytomorphology of adenocarcinoma and reactive, atypical, and benign mesothelial hyperplasia. METHODS The immunohistochemical markers namely EMA, calretinin, vimentin, desmin and Ecadherin were applied on cell blocks employing streptavidin-biotin method. Immunohistochemistry was interpreted by giving scores to the percentage of stained cells. RESULTS EMA and E-cadherin had 100 % sensitivity in diagnosing adenocarcinoma whereas calretinin, vimentin and desmin were 100 % sensitive in diagnosing reactive, atypical mesothelial carcinoma on the cell block preparations. CONCLUSIONS Immunocytochemistry of fluid should be carried out on the cell block preparation where cytological diagnosis on conventional smear and cell block fails to detect malignant cells in the preparation. KEY WORDS Cell Block, Adenocarcinoma, Mesothelial Cells, Immunohistochemistry, EMA, Calretinin, Vimentin, Desmin, E-Cadherin

Perineurioma versus meningioma. A multi-institutional immunohistochemical and ultrastructural studY

ABSTRACT The differential diagnosis between perineurioma (PN) and meningioma (MEN) can be difficult by histology and immunohistochemistry (IHC) because the perineurium and arachnoid have the same embryological origin. However, there are no comparative studies determining conclusive parameters for the differential diagnosis. The aim of this study is to compare IHC of PN and MEN and their ultra-structural characteristics to elucidate which are the useful data that allow differentiate both entities. Thirty-five MEN were analyzed, and 15 PN, (11 skin and soft tissues and four oral cavity). IHC for epithelial membrane antigen (EMA), Claudin-1, GLUT-1, somatostatin-2 receptor (SSTR-2), and progesterone receptor (RP) was performed. Ultrastructural studies were performed on 8 MEN and 15 PN. Only in PN Claudin-1 was positive in 9/11 (90%) cases and GLUT-1 in 7/11 (63%) cases. In MEN, the progesterone receptor was expressed in 21/35 (60%) cases and no case expressed Claudin-1 and GLUT-1; EMA was expressed in all MEN cases and 93% of PN. SSTR-2 was expressed weakly in six cases of MEN (17%), and it was not considered useful for differential diagnosis. On ultrastructure, PN showed thin and parallel processes, some caveolae, and lacked cell junctions. The cellular processes were surrounded by a collagenous stroma in 94% of the cases. MEN were characterized by curved cytoplasmic cell processes showing desmosomes in 75% of cases. Ultrastructural findings aid in the differential diagnosis between PN and MEN, especially if molecular studies are not available.

DNA Flow cytometry and immunohistochemical evaluation of effusion specimens

DNA Flow cytometry and immunohistochemical evaluation of effusion specimens - JDPO- Print ISSN No: - 2581-3714 Online ISSN No:- 2581-3706 Article DOI No:- 10.18231/2581-3706.2019.0004, IP Journal of Diagnostic Pathology and Oncology-IP J Diagn Pathol Oncol

Detection of micrometastasis in axillary lymph nodes of breast carcinoma patients and its association with clinical outcome.

There is heterogeneity in the clinical behavior of breast carcinoma patients with node negativity. Studies have analyzed different factors influencing the outcome in such patients. It is suggested that the presence of nodal micrometastasis can act as a tool in predicting the aggressiveness of these tumors. The aim of this study is to assess the yield of micrometastasis/isolated tumor cell (ITC) by ultrastaging the morphologically negative axillary nodes and staining them with immunohistochemistry for epithelial membrane antigen. The association of such metastasis with the clinical outcome is determined. This was a retrospective analytical study. One hundred cases of node-negative breast carcinoma patients who underwent surgery along with axillary lymph node dissection were selected. The largest node from the axillary dissection was selected and subjected to ultrastaging and immunohistochemical staining (as sentinel node dissection was not a routine practice at that time), to look for occult metastasis in the form of micrometastasis or ITCs. Occurrence of events in the form of recurrence or death was noted. Association of the parameters was analyzed using Fisher's exact test. Among the 100 cases, 79 patients were followed up for a minimum period of 5 years. Two cases had micromets in one node each. These two patients were among the eight, who developed events subsequently (25%). Hence, a statistically significant association was found between the presence of micromets with events. There is a statistically significant association between the presence of micromets and disease recurrence. Hence, we suggest that ultrastaging of the negative axillary node (now sentinel node, as it is being routinely done) might prove effective in predicting the events/prognosis in clinically and morphologically node-negative breast carcinoma patients.

Choroid Plexus Cyst in a Neonatal Burmeister's Porpoise (Phocoena spinipinnis)

Neuroectodermal developmental anomalies are reported rarely in cetaceans and central nervous system cysts are not described. We describe the gross, microscopical, histochemical and immunohistochemical features of a neuraxial myelencephalic cyst in a stranded neonatal Burmeister's porpoise (Phocoena spinipinnis). Grossly, a subdural, extra-axial, well-demarcated, yellow fluid-filled cystic structure (1.9×1.6×1cm) expanded the left foramen of Luschka, the left caudolateral cerebellar recess and the left cranioventral myelencephalon. The cyst displaced the ipsilateral ventral paraflocculus and distended the underlying cranial nerves IX, X, XI and XII. Microscopically, the cystic structure was lined by a monolayer of low cuboidal to flattened epithelium supported by a thin fibrovascular matrix. Immunohistochemistry (IHC) revealed strong and diffuse expression of AE1/AE3 and focal positivity for vimentin. IHC for epithelial membrane antigen, glial fibrillary acid protein, synaptophysin and S100 was negative. Based on these findings, an extra-axial cyst of the choroid plexus of the fourth ventricle (CCPFV) was diagnosed. The pathological relevance of the CCPFV in this case is uncertain. The cause of death involved severe perinatal interspecific (shark) trauma. The present case provides the first evidence of a neuroepithelial cyst in cetacean species.

Identification of Bile Duct Paucity in Alagille Syndrome: Using CK7 and EMA Immunohistochemistry as a Reliable Panel for Accurate Diagnosis.

Bile duct paucity is the absence or marked reduction in the number of interlobular bile ducts (ILBD) within portal tracts. Its syndromic variant, Alagille syndrome (ALGS), is a multisystem disorder with effects on the liver, cardiovascular system, skeleton, face, and eyes. It is inherited as an autosomal dominant trait due to defects in NOTCH signaling pathway. ALGS is characterized by vanishing ILBD with subsequent chronic obstructive cholestasis in approximately 89% of cases. Cholestasis stimulates formation of new bile ductules through a process of neoductular reaction, making it difficult to evaluate the presence or absence of ILBD. Therefore, finding a method to differentiate clearly between ILBD and the ductular proliferation is essential for accurate diagnosis. A database search identified 28 patients with confirmed diagnosis of ALGS between 1992 and 2014. Additionally, 7 controls were used. A panel of two immunostains, cytokeratin 7 (CK7) and epithelial membrane antigen (EMA), was performed. CK7 highlighted the bile duct epithelium of ILBD and ductular proliferation, while EMA stained only the brush border of ILBD. In our ALGS group, the ratio of EMA-positive ILBD to identified portal tracts was 12.6% (range, 0%-41%). However, this same ratio was 95.0% (range, 90%-100%) among control cases (P < 0.001). We propose a panel of two immunostains, CK7 and EMA, to differentiate ILBD from ductular proliferation in patients with cholestasis. With this panel, identification of bile duct paucity can be achieved. Additional studies, including molecular confirmation and clinical correlation, would provide a definitive diagnosis of ALGS.

Immunohistochemical detection of axillary lymph node micrometastases in node negative breast cancer patients using cytokeratin and epithelial membrane antigen.

Background and Objective:The study was conducted to detect occult metastases in lymph node negative breast cancer patients using cytokeratin (CK) and epithelial membrane antigen (EMA) immunohistochemistry (IHC) and correlate this with primary tumor size and grade.Materials and Methods:A total of 32 cases including 12 prospective and 20 retrospective cases of axillary lymph node negative breast cancer were studied. CK and EMA IHC were performed to detect micrometastases.Results:Axillary lymph node metastases were detected in 18.75% of previously node negative cases using CK and EMA IHC. CK was found to be more sensitive for detection of metastases compared to EMA. A highly significant correlation was observed between tumor grade and axillary lymph node metastases detected by CK and EMA. However, no significant correlation was found between tumor size and axillary lymph node metastases detected by IHC.Conclusion:In the present study, there was an increase of 18.75% in the occult metastases detection rate using CK and EMA. To conclude, IHC detection of occult metastases should be done using CK in all axillary node negative cases, especially in T1 and T2 stage tumors.

The histopathological spectrum of cutaneous meningeal heterotopias: clues and pitfalls

To describe the histopathological features of heterotopic cutaneous meningeal tissue. Nineteen cases were collected between 1993 and 2010. Immunohistochemistry for epithelial membrane antigen (EMA), neuron specific enolase (NSE), S100, glial fibrillary acid protein (GFAP), progesterone receptor (PR), CD31, glucose transporter-1 (Glut-1), podoplanin and NKI-C3 was performed. Lesions were congenital (100%) and presented as aplasia cutis with alopecia (63%) or lumps (37%), on the scalp (18 of 19) and sacral region. Resonance magnetic imaging revealed four underlying anomalies of the neuraxis. Histopathological analysis showed meningeal tissue arranged in four variably associated architectural patterns: fibrous (100%), pseudovascular (100%), cellular (68%) and pseudomyxoid (32%). Other features included collagen bodies (58%), calcifications (26%) and dermal melanocytes (32%). Heterotopic brain tissue or heterotopic ependymal cyst was associated in two cases. Arachnoidal cells expressed EMA and NSE, but not S100 protein, CD31 or GFAP. They expressed podoplanin (93%), especially in pseudovascular areas, NKI-C3 (79%), and less frequently Glut-1 (46%) and PR (30%). Histopathological features of cutaneous meningeal heterotopias are various and sometimes misleading. Fibrous lesions should not be misdiagnosed as aplasia cutis. Podoplanin-positive pseudovascular spaces represent the main pitfall and should not be diagnosed as lymphangioma. Correct diagnosis is confirmed by EMA and NSE coexpression within the lesion.

Intracranial clear cell meningioma: a clinicopathologic study of 15 cases

Clear cell meningioma (CCM) is a rare histological variant of meningioma. CCM has a high recurrence rate and aggressiveness. In this study, we reviewed our experience in the treatment of the lesion. Here we present a series of 15 patients with intracranial CCM. The clinical data were retrieved from the records of our Neurosurgery Department and the patients' prognoses were attained by clinic service and telephone. Immunohistochemistry for epithelial membrane antigen (EMA), vimentin, glial fibrillary acidic protein (GFAP), CD10, and S-100 was done, and the MIB-1 labeling index was calculated in all cases. The 15 patients included eight males and seven females; the mean age was 34.8 years. The most frequent initial symptoms were headache and hearing loss. The most common location was the cerebellopontine angle (CPA) zone. Eleven patients had total removal and four patients underwent subtotal removal. Histological features of atypia were present in different proportions, from 6.7% to 100%, and six cases accorded with atypia. Three tumors showed brain invasion. EMA and vimentin were 100% positive, and CD10 was 100% negative. GFAP was 87% negative and S-100 was 93% negative. The mean follow-up period was 36.7 months. Three patients with brain invasion all recurred and five cases with atypia recurred. In 11 patients with total removal, six patients recurred. In four patients with subtotal removal, three patients recurred. Kaplan-Maier analysis showed that incomplete surgical resection was significantly associated with recurrence (p = 0.001). The MIB-1 labeling index for recurrence was 5.7 ± 2.7% versus 2.8 ± 1.5% for no recurrence (p = 0.036). CCM is a rare subtype of meningioma, with a tendency to present in younger patients and a propensity to recur. Immunohistochemistry plays a vital role in differentiating CCM from other tumors. Brain invasion, atypia and MIB-1 labeling index are likely to predict the recurrence. The extent of resection might be connected with the prognosis.

Comparison of Immunohistochemistry with Conventional Histopathology for Evaluation of Sentinel Lymph Node in Breast Cancer

The best method of pathological evaluation of sentinel lymph node in breast cancer has not been agreed upon. Immunohistochemical (IHC) techniques have shown a greater sensitivity over conventional histology for the detection of micrometastais. The aim of the study was to determine whether IHC for Epithelial Membrane Antigen (EMA) on the sentinel node could be more sensitive than conventional histology for diagnosing micrometastasis in sentinel lymph nodes. Eighty-four clinically node negative breast cancer patients underwent sentinel node biopsy at time of surgery for breast cancer. The node was subjected to conventional histopathology as well as IHC for EMA. The sensitivity of histology viz a viz IHC for EMA for detection of sentinel node metastasis was 88% and the specitficity was 96%. The overall diagnostic accuray of histology viz a viz IHC was 93%. There were 4 patients with micrometastasis (<2.0mm), which were positive on IHC but negative on histology. Two patients with poorly differentiated breast cancer had a false negative IHC for EMA result as compared to histology. Immunohistochemistry for Epithelial Membrane Antigen can increase the detection rate of micrometastasis in sentinel lymph node. This can have important bearing on deciding the need of adjuvant systemic therapy. A false negative result for EMA may be seen in patients with poorly differential cancer. Therefore the best policy seems to employ both histopathology and IHC for EMA for the comprehensive evaluation of sentinel lymph node in breast cancer.

Dural based primary osteosarcoma in right fronto-temporal region:Case report with review of literature

We report a case of primary dural based osteosarcoma in the right fronto-temporal convexity in a 43-year-old female who presented with a short history of seizure and headache. Radiologic evaluation revealed a well defined brightly enhancing extra-axial lesion in the right fronto-temporal region with a dural tail around the sylvian fissure. The overlying bone was uninvolved. Paraffin section of the tumor showed plump cells with moderate nuclear and cellular pleomorphism with eosinophilic extracellular material (osteiod) between the cells. At a few places, lace like osteiod was seen encasing individual cells signifying osteiod being formed by tumor cells. Immunohistochemistry for EMA was focally positive and negative for S-100 protein and GFAP. A final histopathological diagnosis of dural based primary osteosarcoma of the right fronto-temporal region was rendered. To the best of our knowledge this will be the eighth such case in literature.

A report of supratentorial leptomeningeal hemangioblastoma and a literature review

Hemangioblastomas of the CNS are solid or cystic vascular-rich tumors, most common in the cerebellum, less frequent in the brainstem or spinal cord, and rare in supratentorial locations with meningeal involvement. We document a case in a 58-year-old man who presented with about 2 months history of motor weakness and speech dysfunction. The tumor was a heterogeneously enhanced dural-based tumor with high vascularity and perifocal edema in the left frontal lobe. The tumor was completely removed followed by embolization and preoperative radiotherapy. Histologic examination revealed a hemangioblastoma with features resembling angiomatous meningioma. Immunohistochemistry for epithelial membrane antigen (EMA) and S100 may be helpful to make differential diagnosis. Electron microscopic investigation is essential to differentiate between meningiomas and other leptomeningeal tumors.

Usefulness of epithelial membrane antigen (EMA) to discriminate between perineural invasion and perineural indentation in prostatic carcinoma

Perineural invasion (PNI) is one of the few unequivocal criteria to diagnose adenocarcinoma of the prostate. Distinguishing PNI from perineural indentation (PNIn), however, is sometimes difficult. The aim of this study was to discriminate between PNI and PNIn using EMA immunohistochemistry. We selected representative sections from 87 prostatectomies with prostatic adenocarcinoma. Normal peripheral nerves were continuously encircled with perineurium, which was immunoreactive for EMA. We identified 1319 PNI by carcinomas, 368 PNIn by carcinomas, and 303 PNIn by benign glands. We categorized the EMA immunoreactivity patterns into three classes: samples that displayed discontinuity or complete loss of the perineurium (Type A), samples where there were carcinomas or benign glands in the perineural space or peripheral nerves (Type B) and samples that showed no changes in the perineurium (Type C). For PNI we observed Type A, Type B, and Type C patterns in 55.3%, 24.8% and 19.9% of carcinomas, respectively. The incidence of each of those patterns in PNIn by carcinomas was 32.1%, 14.9% and 53.0%, respectively. Cases of PNIn by benign glands showed Type A or Type C patterns. They did not, however, exhibit Type B patterns. EMA immunostaining will aid the diagnosis of prostatic adenocarcinoma.

Canine biphasic synovial sarcoma: case report and immunohistochemical characterization

The clinical, radiological and pathologic features of a biphasic synovial sarcoma in the left elbow joint of a two-year-old male Rottweiler are presented. The tumor showed positive immunoreactivity for vimentin, Epithelial Membrane Antigen (EMA), p53 and PCNA, while it was negative for the cytokeratin used, S-100, Rb and p21. Immunohistochemistry for EMA allowed the identification of epithelioid components of synovial sarcoma, and may, therefore, contribute in establishing a diagnosis of biphasic synovial sarcoma. Intratumoral variation in PCNA immunoreactivity was minimal, indicating that the various tumor components proliferate at more or less similar rates. Overall, the characterized immunohistochemical profile for canine synovial sarcoma, not defined previously, may provide clues to the histogenesis of the phenotypically mesenchymal and epithelial elements of the tumor, and may be of value in the differential diagnosis of challenging cases, decreasing the risk of under- and mis-diagnosis. Although more cases need to be studied to determine whether there is a consistent pattern of immunostaining in canine synovial sarcoma, its potential significance is discussed in relation to the histogenesis, molecular pathology and differential diagnosis of canine synovial sarcoma.

Cytokeratin 7 and 20 expression in epithelioid sarcoma.

Epithelioid sarcoma (ES) is a rare malignant soft tissue tumor of uncertain histogenesis that arises predominantly in the extremities of young adults. Immunohistochemically, the neoplastic cells are typically positive for vimentin, low molecular weight cytokeratin (CAM5.2) and epithelial membrane antigen (EMA). We examined eight cases of ES from seven different patients. All cases were studied with immunohistochemistry for EMA, CAM5.2 (keratin 8 and 18), 34BE12 (keratins 1, 5, 10 and 14/15), cytokeratins 7 and 20 (CK7, CK20), and CD34. The average patient age was 53 (range 43-76) and the male:female ratio was 5:2. The location was the upper extremity in five tumors, the lower extremity, the perineum, and the paraspinal soft tissue in one tumor each. All cases contained predominantly epithelioid cells, but spindle cells were also present in three cases. All cases contained areas of geographic necrosis. CAM5.2 was strongly positive in seven tumors and focally positive in one (8/8). EMA was diffusely positive in two cases and focally positive in five cases (7/8). CD34 was diffusely positive in 3/8 cases. 34BE12 was diffusely positive in one case and focally positive in two others (3/8). CK7 was diffusely positive in one case and focally positive in another (2/8). CK20 was negative in all cases (0/8). All cases tested were positive for vimentin (6/6), 2 cases were focally positive for HHF35 (2/5), and all cases tested were negative for S-100 protein (0/7). In addition to the known immunoreactivity for CAM5.2 and EMA, there is positivity for CK7 and 34BE12 in a small proportion of cases. None of the cases expressed CK20. This immunophenotypic profile suggests that ES is more similar to carcinoma and synovial sarcoma than to other soft tissue tumors, and may be of diagnostic utility.

Cell renewal and functional morphology of human lactating breast: presentation of a new mode of cell death ('magentosis') characterized by nuclear periodic acid-Schiff reactivity.

Cell kinetics and functional morphology of the human lactating breast were analyzed using 21 paraffin-embedded specimens of the lactating breast obtained by biopsy or surgery and two of the 'resting' breast. Four types (I-IV) of lactating lobules were categorized histologically, and were well correlated with their functional status demonstrated by immunohistochemistry for epithelial membrane antigen, beta-casein, lipase, lactoferrin, secretory component and IgA. Type I corresponded to a pre-lactating state, type II to an actively lactating state, type III to an early stage of regression, and type IV to an advanced stage of regression. Cell proliferation monitored by Ki-67 (MIB-1) immunostaining was at the highest level in type I lobules followed by type IV, while the labeling indices were low in types II and III. Apoptosis demonstrated by terminal deoxynucleotidyl transferase (TdT)-mediated deoxyuridine triphosphate (dUTP)-biotin nick end-labeling (TUNEL) method showed labeling profiles comparable with the proliferative activity. Holocrine-type desquamative cells with intact-appearing nuclei were frequently recognized in type II lobules. In type III, a peculiar form of cell death, designated 'magentosis', was demonstrated. These degenerative cells occasionally shedding into the acinar lumen possessed homogeneous pale, TUNEL-negative nuclei with diastase-resistant periodic acid-Schiff (PAS) reactivity. 'Magentosis' specifically identified in an early phase of involution of the human lactating breast might represent a unique mode of cell death distinguishable from apoptosis and necrosis.

Alcian blue and epithelial membrane antigen are useful markers in differentiating benign from malignant papillae in thyroid lesions

Immunohistochemistry for epithelial membrane antigen (EMA) and histochemistry for alcianophilic substances were performed in 17 cases of papillary thyroid carcinoma (PTC) and 11 cases of benign thyroid lesions showing papillary changes (7 diffuse hyperplastic goitres-Graves' disease; 4 colloid cystic goitres). In all PTCs the glycocalix of the cells lining the papillary structures was strongly positive with anti-EMA antiserum. Alcian blue pH 2.5 stain (AB 2.5) was also positive in 15 of these cases. In contrast, no cases of benign thyroid lesions showed AB 2.5 positivity in the cells lining the papillary structures and the positivity with anti-EMA antiserum, present in only 5 out the 11 cases, was focal and very weak. These results indicate that the presence and distribution of EMA and alcianophilic substances may be useful in distinguishing benign from malignant thyroid lesions containing papillae.