Sources/Clones
Accurate (E13, CCH2), American Research Products (1692-18), Axcel (CCH2), Biodesign (084, BM204, BM219, polyclonal), Biogenesis (BM204, polyclonal), Biogenex (BM204, polyclonal), Chemicon, Dako (AAC10, CCH2), EY Labs, Fitzgerald (M2103126, M210312), International Enzymes (polyclonal), Seralab (E13) and Zymed (DDG9/CCH2).
Fixation/Preparation
These antibodies are suitable for immunohistochemical staining of paraffin-embedded tissue sections. Enzymatic predigestion with trypsin or pepsin is required for clone CCH2. These antibodies may also be used to detect CMV early nuclear proteins in infected human embryonic fibroblasts 24 h following inoculation of clinical specimens on cell culture.
Background
The CCH2 clone recognizes a 43 kD protein, whilst the DDG9 clone recognizes a 76kD protein, both having been demonstrated in glycine-extracted CMV antigen. These proteins are expressed in the immediate early and early stage of CMV replication in infected cells (Zweygberg et al, 1986). Early viral proteins are expressed in the nucleus of infected cells within 6-24 h of infection and prior to viral DNA replication. Several late viral proteins may be demonstrated in the nucleus and the cytoplasm of infected cells. The different viral proteins can be demonstrated in infected cell cultures as well as in infected tissue (Swenson & Kaplan, 1985). These antibodies do not crossreact with adenoviruses or other herpesviruses.
Applications
These antibodies to CMV demonstrate the virus in infected cells, producing a nuclear immunopositive reaction. However, at a later stage, both a nuclear and cytoplasmic immunoreaction with the early CMV antigen is produced, especially with the Zymed product. Antibodies to CMV have a wide application to diagnostic surgical pathology, especially when characteristic CMV inclusions are not clearly evident. CMV infection (latent or active) may be seen in salivary glands, lungs, kidneys, GIT and lymph nodes. Awareness of CMV as an opportunistic infection in the context of immunosuppression suggests the use of CMV immunohistochemistry for definitive diagnosis (Schwartz & Wilcox, 1992). Recently, CMV esophagitis has been observed as a florid aggregate of macrophages without typical inclusions (Greenson, 1997). Small biopsy specimens with such a morphological picture warrant further immunohistochemical study to identify CMV. Conversely, chemotherapy toxicity may mimic CMV gastritis, necessitating CMV immunohistochemistry to exclude false positives (Canioni et al, 1995). Antibodies to CMV may also be applied for the identification of atypical CMV inclusions in gastrointestinal mucosal biopsy specimens, where classic inclusions are rarely found (Schwartz & Wilcox, 1992). The proper recognition of CMV-infected cells in the context of immunosuppression is critical, so that effective therapy is not delayed, preventing further viral dissemination.
Comments
It has been shown that immunohistochemistry with CCH2 detects a higher number of CMV-infected cells than in situ hybridization (Niedobitek et al, 1988). Hence, for routine diagnostic purposes at least, CMV immunohistochemistry would appear to be the method of choice for a rapid, sensitive and specific method of CMV detection.
References
•Canioni D, Vassal G, Donadieu J, Hubert PH, Brousse N 1995. Toxicity induced by chemotherapy mimicking cytomegalovirus gastritis. Histopathology 26: 473-475.
•Greenson JK 1997. Macrophage aggregates in cytomegalovirus esophagitis. Human Pathology 28: 375-378.
•Niedobitek G, Finn T, Herbst H et al 1988. Detection of cytomegalovirus by in situ hybridisation and immunohistochemistry using new monoclonal antibody CCH2: a comparison of methods. Journal of Clinical Pathology 41: 1005-1009.
•Schwartz DA, Wilcox CM 1992. Atypical cytomegalovirus inclusions in gastrointestinal biopsy specimens from patients with the acquired immunodeficiency syndrome: diagnostic role of in situ nucleic acid hybridization. Human Pathology 23: 1019-1026.
•Swenson PD, Kaplan MH 1985. Rapid detection of cytomegalovirus in cell culture by indirect immunoperoxidase staining with monoclonal antibody to an early nuclear antigen. Journal of Clinical Microbiology 21:669-673.
•Zweygberg WB, Wirgart B, Grillner L 1986. Early detection of cytomegalovirus in cell culture by a monoclonal antibody. Journal of Virological Methods 14: 65-69.
Bibliografía
Manual of diagnostic antibodies for immunohistology / Anthony S.-Y. Leong, Kumarasen Cooper, F. Joel W.-M. Leong.
Accurate (E13, CCH2), American Research Products (1692-18), Axcel (CCH2), Biodesign (084, BM204, BM219, polyclonal), Biogenesis (BM204, polyclonal), Biogenex (BM204, polyclonal), Chemicon, Dako (AAC10, CCH2), EY Labs, Fitzgerald (M2103126, M210312), International Enzymes (polyclonal), Seralab (E13) and Zymed (DDG9/CCH2).
Fixation/Preparation
These antibodies are suitable for immunohistochemical staining of paraffin-embedded tissue sections. Enzymatic predigestion with trypsin or pepsin is required for clone CCH2. These antibodies may also be used to detect CMV early nuclear proteins in infected human embryonic fibroblasts 24 h following inoculation of clinical specimens on cell culture.
Background
The CCH2 clone recognizes a 43 kD protein, whilst the DDG9 clone recognizes a 76kD protein, both having been demonstrated in glycine-extracted CMV antigen. These proteins are expressed in the immediate early and early stage of CMV replication in infected cells (Zweygberg et al, 1986). Early viral proteins are expressed in the nucleus of infected cells within 6-24 h of infection and prior to viral DNA replication. Several late viral proteins may be demonstrated in the nucleus and the cytoplasm of infected cells. The different viral proteins can be demonstrated in infected cell cultures as well as in infected tissue (Swenson & Kaplan, 1985). These antibodies do not crossreact with adenoviruses or other herpesviruses.
Applications
These antibodies to CMV demonstrate the virus in infected cells, producing a nuclear immunopositive reaction. However, at a later stage, both a nuclear and cytoplasmic immunoreaction with the early CMV antigen is produced, especially with the Zymed product. Antibodies to CMV have a wide application to diagnostic surgical pathology, especially when characteristic CMV inclusions are not clearly evident. CMV infection (latent or active) may be seen in salivary glands, lungs, kidneys, GIT and lymph nodes. Awareness of CMV as an opportunistic infection in the context of immunosuppression suggests the use of CMV immunohistochemistry for definitive diagnosis (Schwartz & Wilcox, 1992). Recently, CMV esophagitis has been observed as a florid aggregate of macrophages without typical inclusions (Greenson, 1997). Small biopsy specimens with such a morphological picture warrant further immunohistochemical study to identify CMV. Conversely, chemotherapy toxicity may mimic CMV gastritis, necessitating CMV immunohistochemistry to exclude false positives (Canioni et al, 1995). Antibodies to CMV may also be applied for the identification of atypical CMV inclusions in gastrointestinal mucosal biopsy specimens, where classic inclusions are rarely found (Schwartz & Wilcox, 1992). The proper recognition of CMV-infected cells in the context of immunosuppression is critical, so that effective therapy is not delayed, preventing further viral dissemination.
Comments
It has been shown that immunohistochemistry with CCH2 detects a higher number of CMV-infected cells than in situ hybridization (Niedobitek et al, 1988). Hence, for routine diagnostic purposes at least, CMV immunohistochemistry would appear to be the method of choice for a rapid, sensitive and specific method of CMV detection.
References
•Canioni D, Vassal G, Donadieu J, Hubert PH, Brousse N 1995. Toxicity induced by chemotherapy mimicking cytomegalovirus gastritis. Histopathology 26: 473-475.
•Greenson JK 1997. Macrophage aggregates in cytomegalovirus esophagitis. Human Pathology 28: 375-378.
•Niedobitek G, Finn T, Herbst H et al 1988. Detection of cytomegalovirus by in situ hybridisation and immunohistochemistry using new monoclonal antibody CCH2: a comparison of methods. Journal of Clinical Pathology 41: 1005-1009.
•Schwartz DA, Wilcox CM 1992. Atypical cytomegalovirus inclusions in gastrointestinal biopsy specimens from patients with the acquired immunodeficiency syndrome: diagnostic role of in situ nucleic acid hybridization. Human Pathology 23: 1019-1026.
•Swenson PD, Kaplan MH 1985. Rapid detection of cytomegalovirus in cell culture by indirect immunoperoxidase staining with monoclonal antibody to an early nuclear antigen. Journal of Clinical Microbiology 21:669-673.
•Zweygberg WB, Wirgart B, Grillner L 1986. Early detection of cytomegalovirus in cell culture by a monoclonal antibody. Journal of Virological Methods 14: 65-69.
Bibliografía
Manual of diagnostic antibodies for immunohistology / Anthony S.-Y. Leong, Kumarasen Cooper, F. Joel W.-M. Leong.
