A comparative evaluation of immunohistochemical markers for the differential diagnosis between malignant mesothelioma, non-small cell carcinoma involving the pleura, and benign reactive mesothelial cell proliferation
ABSTRACT Histopathological diagnosis of malignant mesothelioma (MM) and differentiating it from tumors infiltrating the pleura is very difficult. Distinguishing benign reactive mesothelial cell proliferation from MM also presents problems. The objective of this study was to evaluate the significance of selected immunohistochemical stains in differentiating MM from non-small cell lung cancers infiltrating the pleura and from benign reactive mesothelial cell proliferation.
The material encompassed 86 cases of MM, 54 cases of NSCLC infiltrating the pleura, and 43 cases of benign reactive mesothelial cell proliferation. The MM cases were reclassified according to the WHO criteria (2004): epithelioid, 61 cases (71%), including well-differentiated papillomatous, 3 cases; sarcomatous, 6 cases (6.8%); fibrous, 4 cases (4.7%); biphasic, 15 cases (17.5%). A panel of immunohistochemical stains was used in this study. It included broad-spectrum antibodies to cytokeratins (CKAE1/AE3, CKMNF116), vimentin, epithelial membrane antigen (EMA), mesothelial cells (HBME1, CK5/6, calretinin), adenocarcinoma cells (BerEp4, B72.3, CEA, TTF1), antibodies enabling the assessment of proliferation (Mib1) and cell-cycle regulating proteins (p53).
Coexpression of cytokeratins and vimentin was found in 63.9% of MM cases and cell-membrane reactions with EMA were seen in 58.9%. Positive staining for HBME1, CK5/6, calretinin, BerEp4, B72.3, CEA and p53 was obtained in 76.7%, 51.2%, 66.7%, 1.2%, 6.2%, 1.2% and 51% of the cases, respectively. None of the MM cases stained for TTF1. MM by WHO subgroups: Coexpression of cytokeratins and vimentin occurred in 55.7% cases of epithelioid MM, 93.3% of biphasic MM, 66.6% of sarcomatous MM, and in 100% of fibrous MM cases. Positive staining for HBME1, CK5/6, and calretinin was seen only in the epithelioid and mixed subtypes of MM; the respective percentages of positive reactions were: HBME1, 90.2% and 73.3%; CK5/6 58.2% and 53.3%; calretinin, 72% and 75%. Non-small cell lung cancers infiltrating the pleura: Coexpression of cytokeratin and vimentin was found in 17.6% of the cases, positive staining of membranes for EMA, in 13% cases. Positive staining for HBME1 was observed in 22.6% of the cases, for CK5/6, in 9.3%, for calretinin, in 2%, for BerEp4, in 72.2%, for B72.3, in 64.1%, for CEA, in 58.5%, and for TTF1, in 43.8%. Benign reactive mesothelial cell proliferation: Protein p53 was present in 9.3% of cases, whereas no positive staining for EMA was found. Differentiation of MM from non-small cell carcinomas: Among the antibodies used in the study, anti-HBME1 had the highest sensitivity (76.7%) but lowest specificity (77.4%). Staining for calretinin showed high specificity (99.8%), as did CEA and TTF1 (98.8% and 100%), with moderate sensitivity (66.7%, 58.5% and 43.8%, respectively). BerEp4 showed the highest sensitivity (72.2%) and specificity (98.8%).
In diagnosing mesothelioma it is necessary to use a panel of immunohistochemical stains, which should contain antibodies to markers for adenocarcinoma and mesothelioma. Due to the high costs of such a study, a two-stage method is advantageous. The best combination of sensitivity and specificity was found for BerEp4, CEA, and TTF1 and for calretinin and HBME1. In the diagnosis of spindle-cell pleural tumors and the fibrous form of MM and benign reactive mesothelial cell proliferation , markers of mesothelial cells are noncontributory. Immunohistochemical staining fails to identify a reactive process, but a diffuse, positive stain for EMA and the presence of protein p53 support the diagnosis of MM.
- SourceAvailable from: Rafał Sapierzyński
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- "Despite undeniable usefulness of the routine cytopathological examination in the majority of cases, cytopathological diagnosis cannot always be completed , especially in some specific types of lesions. Differentiation between some neoplastic and non-neoplastic hyperplastic processes (epithelial dysplasia , connective tissue hyperplasia, postinflammatory epithelial changes) can be difficult or even impossible based on cytopathology only (Stepien et al. 2000, Geninet et al. 2003, Sato et al. 2005, Ordonez 2006, Szczepulska-Wójcik et al. 2007, Bertazzolo et al. 2012). Some additional diagnostic methods, such as the surgical biopsies of lesions or specialized immunohistochemical (IHC) or immunocytochemical (ICC) staining, that are required in differential diagnosis of tumors within serosal cavities and are considered to be a " gold standard " , are not readily available in veterinary medicine at present (Bertazzolo et al. 2012). "
ABSTRACT: The presence of tumor within the serosal cavities, often connected with accumulation of serosal effusion, is a quite common problem in the small animal veterinary medicine. The first step in diagnosis of such cases is cytopathological examination. The aim of the present study was to evaluate the usefulness of cytology and immunocytochemistry, using commercially available antibodies (anti-cytokeratin, anti-vimentin, and anti-desmin), in differential diagnosis of malignant tumors located within serosal cavities in dogs. The final cytological diagnosis of carcinoma/adenocarcinoma, sarcoma, and mesothelioma was obtained on the basis of routine cytopathology and immunocytochemistry, and then confirmed by histopathology and immunohistochemistry. Cytoplasmic immunoreactivitiy of normal mesothelid cells and cytoplasmic immunoreactivity of hyperplastic mesothelial cells revealed constant and strong expression of all examined intermediate filaments: cytokeratin, vimentin and desmin. Application of routine cytopathology and immunocytochemistry allowed 32 neoplastic tumors to be detected: 19 cases of carcinomas/adenocarcinomas, 6 cases of sarcomas, 7 cases of mesotheliomas. Immunostaining of cytopathological samples with chosen set of antibodies: anti-cytokeratin, anti-vimentin, anti-desmin is a useful, and low invasive test for differentiation between mesotheliomas and carcinomas/adenocarcinomas in dogs.Polish journal of veterinary sciences 04/2014; 17(1):149-59. DOI:10.2478/pjvs-2014-0020 · 0.60 Impact Factor
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ABSTRACT: Differentiating reactive mesothelial cells (RMs) from metastatic adenocarcinoma cells (MAC) in serous fluids based on cytomorphologic features alone can be very challenging. Various immunocytochemical (ICC) markers have been used to maximize the diagnostic accuracy, however, cytopathologists still encounter difficulties in effusion cytologic diagnosis. The aim of this study was to evaluate previous and recent ICC stains to identify the most sensitive and specific markers and the best panel for differentiating RM from MAC. Cell block sections from 41 MAC and 43 RM effusions cases were subjected to ICC staining for MOC-31, BerEp4, carcinoembryonic antigen (CEA), calretinin, HBME-1, CK5/6, and D2-40. For the MAC cases, the sensitivity of BerEp4, MOC-31, and CEA was 82.9, 92.6, and 17%, respectively, and the specificity was 95.3, 93, and 100%, respectively. For the RM cases, the sensitivity of calretinin, CK5/6, D2-40, and HBME-1 was 95.3, 27.9, 58.1, and 93%, respectively, and the specificity was 70.7, 73.1, 75.6, and 82.9%, respectively. The results show that BerEp4 and MOC-31 are highly sensitive and specific for detecting MAC, whereas calretinin and HBME1 are highly sensitive but only modestly specific for detecting RM cases (P < 0.05). Forced entry logistic regression revealed that using MOC-31, BerEp4, HBME-1, and calretinin, is an excellent panel for making correct diagnosis with 97.6% sensitivity in detecting MAC and 90.7% specificity in detecting RM. We conclude that adding a panel of MOC-31, BerEp4, calretinin, and HBME-1 immunostains to routine cytomorphologic features can greatly enhance the diagnostic accuracy of serous effusions.Diagnostic Cytopathology 05/2009; 37(5):324-32. DOI:10.1002/dc.21006 · 1.12 Impact Factor
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ABSTRACT: Both pathologists and clinicians are challenged by the diagnosis of a particular variant of the peripheral adenocarcinoma with involvement of the pleura parietalis, the so-called pseudo-mesotheliomatous adenocarcinoma of the lung (PMAC), which is hard to differentiate from epithelioid mesothelioma on imaging and cytology, macroscopically as well as histologically. However, the exact diagnosis is not only crucial for the patient's therapy but also for insurance matters. Immunohistochemical evaluation represents a quick and a relatively cheap tool for which a few antibody panels have been proposed in recent years as being suitable to distinguish between these two entities. One of the positive markers for epithelioid mesothelioma most often suggested seems to be calretinin. We would like to report on a case of PMAC with the special feature of positive calretinin immunohistochemical staining. Using histochemistry and a few additional antibodies we were able to reliably characterize the tumor and provide the patient with appropriate therapy. This article gives a short overview of the possibilities available for distinguishing between these two entities in the context of a case report.Der Pathologe 10/2009; 31(4):283-9. · 0.39 Impact Factor