Sources/Clones
Axcel/Accurate, Dako and Diagnostic Bioscience.
Fixation/Preparation
Ber-EP4 can be used on formalin-fixed, paraffin-embedded tissue sections. Prolonged formalin fixation can be deleterious to immunoreactivity, which is enhanced by HIER or by enzymatic predigestion with proteolytic enzymes such as trypsin and pronase. Ber-EP4 may also be used to label acetone-fixed cryostat sections and fixed-cell smears. A major advantage of this antibody is the high sensitivity which allows it to be used at high dilutions.
Background
Ber-EP4 was raised against MCF-7 cells and is directed against two glycoproteins of 34 and 49 kD present on the surface and in the cytoplasm of all epithelial cells with the exception of the superficial layers of squamous epithelia, hepatocytes and parietal cells (Latza et al, 1990). Although it is not yet clear what antigen is recognized by the antibody, an absence of reactivity to keratins was found in immunoblotting experiments. A positive reaction is seen in epithelial cells known to contain large amounts of the Ber-EP4 antigen, e.g. epithelial cells in the bile ducts and ducts of the epididymis.
Applications
Ber-EP4 shows a broad pattern of reactivity with human epithelial tissues from simple epithelia to basal layers of stratified non-keratinized squamous epithelium and epidermis (Appendix 1.20). In addition' most cases of carcinoma demonstrated immunoreaction with this antibody (Latza et al, 1990). However, two cases of malignant mesothelioma studied reacted negatively. In a separate study (Sheibani et al, 1992), 87% of 83 adenocarcinomas were found to express Ber-EP4. The only adenocarcinomas that failed to react were of breast origin (eight of 25 cases non-reactive) and kidney (all three cases non-reactive). In contrast, only one of 115 mesotheliomas studied showed positivity.
Focal expression of Ber-EP4 in the mesothelium of the peritoneum and the ovarian surface epithelium adjacent to endometriotic lesions suggests that the mesothelium possibly acquires characteristics of epithelial nature, supporting a metaplastic process of the peritoneal mesothelium in the pathogenesis of endometriosis (Nakayama et al, 1994).
Comments
Any attempt to use Ber-EP4 to help distinguish epithelial mesothelioma from adenocarcinoma should be accompanied by a panel of antibodies (Appendix 1.16) including CEA, Leu-M1, B72.3 (all three antibodies in combination were reported to distinguish over 90% pulmonary adenocarcinomas from pleural mesotheliomas; Sheibani et al, 1991) and calretinin. In addition, anti-EMA has been shown to produce a distinctive pattern of membrane staining corresponding to the circumferential long microvilli which are pathognomonic of malignant mesothelial cells (Leong et al, 1990).
References
•Latza U, Niedobitek G, Schwarting R et al 1990. Ber-EP4: new monoclonal antibody which distinguishes epithelia from mesothelia. Journal of Clinical Pathology 43: 213-219.
•Leong AS-Y, Parkinson R, Milios J 1990. "Thick" cell membranes revealed by immunocytochemical staining: a clue to the diagnosis of staining: a clue to the diagnosis of mesothelioma. Diagnostic Cytopathology 6: 9-13.
•Nakayama K, Masuzawa H, Shuan-Fang L et al 1994. Immunohistochemical analysis of the peritoneum adjacent to endometriotic lesions using antibodies for Ber-EP4 antigen, estrogen receptors and progesterone receptors: implication of peritoneal metaplasia in the pathogenesis of endometriosis. International Journal of Gynecologic Pathology 13: 348-358.
•Sheibani K, Shin S, Kezirian J et al 1991. Ber-EP4 antibody as a discriminant in the differential diagnosis of malignant mesothelioma versus adenocarcinomas. American Journal of Surgical Pathology 15: 779-784.
•Sheibani K, Esteban JM, Bailey A, Battifora H, Weiss L 1992. Immunologic and molecular studies as an aid to the diagnosis of malignant mesothelioma. Human Pathology 23: 107-116.
Bibliografia
Manual of diagnostic antibodies for immunohistology / Anthony S.-Y. Leong, Kumarasen Cooper, F. Joel W.-M. Leong.
Axcel/Accurate, Dako and Diagnostic Bioscience.
Fixation/Preparation
Ber-EP4 can be used on formalin-fixed, paraffin-embedded tissue sections. Prolonged formalin fixation can be deleterious to immunoreactivity, which is enhanced by HIER or by enzymatic predigestion with proteolytic enzymes such as trypsin and pronase. Ber-EP4 may also be used to label acetone-fixed cryostat sections and fixed-cell smears. A major advantage of this antibody is the high sensitivity which allows it to be used at high dilutions.
Background
Ber-EP4 was raised against MCF-7 cells and is directed against two glycoproteins of 34 and 49 kD present on the surface and in the cytoplasm of all epithelial cells with the exception of the superficial layers of squamous epithelia, hepatocytes and parietal cells (Latza et al, 1990). Although it is not yet clear what antigen is recognized by the antibody, an absence of reactivity to keratins was found in immunoblotting experiments. A positive reaction is seen in epithelial cells known to contain large amounts of the Ber-EP4 antigen, e.g. epithelial cells in the bile ducts and ducts of the epididymis.
Applications
Ber-EP4 shows a broad pattern of reactivity with human epithelial tissues from simple epithelia to basal layers of stratified non-keratinized squamous epithelium and epidermis (Appendix 1.20). In addition' most cases of carcinoma demonstrated immunoreaction with this antibody (Latza et al, 1990). However, two cases of malignant mesothelioma studied reacted negatively. In a separate study (Sheibani et al, 1992), 87% of 83 adenocarcinomas were found to express Ber-EP4. The only adenocarcinomas that failed to react were of breast origin (eight of 25 cases non-reactive) and kidney (all three cases non-reactive). In contrast, only one of 115 mesotheliomas studied showed positivity.
Focal expression of Ber-EP4 in the mesothelium of the peritoneum and the ovarian surface epithelium adjacent to endometriotic lesions suggests that the mesothelium possibly acquires characteristics of epithelial nature, supporting a metaplastic process of the peritoneal mesothelium in the pathogenesis of endometriosis (Nakayama et al, 1994).
Comments
Any attempt to use Ber-EP4 to help distinguish epithelial mesothelioma from adenocarcinoma should be accompanied by a panel of antibodies (Appendix 1.16) including CEA, Leu-M1, B72.3 (all three antibodies in combination were reported to distinguish over 90% pulmonary adenocarcinomas from pleural mesotheliomas; Sheibani et al, 1991) and calretinin. In addition, anti-EMA has been shown to produce a distinctive pattern of membrane staining corresponding to the circumferential long microvilli which are pathognomonic of malignant mesothelial cells (Leong et al, 1990).
References
•Latza U, Niedobitek G, Schwarting R et al 1990. Ber-EP4: new monoclonal antibody which distinguishes epithelia from mesothelia. Journal of Clinical Pathology 43: 213-219.
•Leong AS-Y, Parkinson R, Milios J 1990. "Thick" cell membranes revealed by immunocytochemical staining: a clue to the diagnosis of staining: a clue to the diagnosis of mesothelioma. Diagnostic Cytopathology 6: 9-13.
•Nakayama K, Masuzawa H, Shuan-Fang L et al 1994. Immunohistochemical analysis of the peritoneum adjacent to endometriotic lesions using antibodies for Ber-EP4 antigen, estrogen receptors and progesterone receptors: implication of peritoneal metaplasia in the pathogenesis of endometriosis. International Journal of Gynecologic Pathology 13: 348-358.
•Sheibani K, Shin S, Kezirian J et al 1991. Ber-EP4 antibody as a discriminant in the differential diagnosis of malignant mesothelioma versus adenocarcinomas. American Journal of Surgical Pathology 15: 779-784.
•Sheibani K, Esteban JM, Bailey A, Battifora H, Weiss L 1992. Immunologic and molecular studies as an aid to the diagnosis of malignant mesothelioma. Human Pathology 23: 107-116.
Bibliografia
Manual of diagnostic antibodies for immunohistology / Anthony S.-Y. Leong, Kumarasen Cooper, F. Joel W.-M. Leong.
