Sources/Clones
Accurate (CS1-4), Biodesign (polyclonal), Dako (CS1-4), EY Labs, Novocastra (polyclonal) and Research Diagnostics (4C5).
Fixation/Preparation
Applicable to formalin-fixed, paraffin-embedded tissue sections. Enzymatic digestion (e.g. trypsin) is essential to enhance immunopositivity. The application of microwave irradiation for antigen retrieval has also been used to good effect (Kaczorowski et al, 1994). This antibody may also be used for labeling acetone-fixed cryostat sections or fixed-cell smears.
Background
The antibody (isotype: IgG1, k) has been raised against recombinant fusion protein containing sequences of bacterialb -galactosidase and the EBV-encoded latent membrane protein (LMP-1). LMP is one of the few viral proteins that are expressed in a latent infection. The antibody reacts with a 60 kD latent membrane protein encoded by the BNLF gene of the Epstein-Barr virus. Being a cocktail of clones CS1, CS2, CS3 and CS4, all four anti-LMP antibodies recognize distinct epitopes on the hydrophilic carboxyl region of LMP (Rowe et al, 1987). These four epitopes are present on the internal aspect of the membrane-associated viral LMP. Therefore the antibody does not react with viable cells, but with fixed cells in paraffin sections, cytological preparations and cryostat sections and in immunoblotting.
Applications
The antibody is characterized by its strong positivity with EBV+ lymphoblastoid cell lines and EBV-infected B-cell immunoblasts in infectious mononucleosis. Although EBV is consistently present in nasopharyngeal undifferentiated carcinoma among Oriental patients, LMP-1 antibody is only positive in about 60% of cases (Hording et al, 1993; Lopategui et al, 1994). LMP protein expression is especially useful in identifying these cancers in cervical lymph node metastases. This antibody may also be useful in the diagnosis of lymphoepithelioma-like carcinoma of the lung, mediastinum, stomach and paranasal sinuses (Dimery et al, 1988; Weiss et al, 1989; Shibata et al, 1991).
Posttransplantation lymphoproliferative disorders arising in patients treated with a variety of immunosuppressive regimens after organ transplantation usually show a type III latency pattern with LMP-1 expression (Delecluse et al, 1995). The EBV+ AIDS-associated B-cell lymphomas usually demonstrate a latency type III in the large cell lymphomas, permitting the use of antibody to LMP-1 (Hamilton-Du Toit et al, 1993).
Nasal T/NK-cell lymphoma is strongly associated with EBV (Kanavaros et al, 1993). However, LMP-1 protein expression has been inconsistent on paraffin sections, although one study consistently demonstrated LMP-1 protein in frozen sections, suggesting a low level of protein expression. LMP-1 immunohistochemistry is positive in 17% of adult T-cell leukemia/lymphoma (Tokunaga et al, 1993). LMP-1 expression has also been associated with an aggressive clinical course and hepatosplenomegaly in nodal T-cell lymphomas (Bruin, 1993). About 20-30% of CD 30 (Ki-1)+ anaplastic large cell lymphoma show LMP-1 immunoreaction (Herbst et al, 1991).
Approximately 50% of Hodgkin's disease cases are associated with EBV. In almost all of these positive cases, nearly all the Reed-Sternberg cells are positive for EBV. Using modern epitope retrieval techniques, an almost 1:1 correlation between the results of LMP-1 paraffin-based immunohistochemistry studies and EBER in situ hybridization studies has been demonstrated in Reed-Sternberg cells and Hodgkin's cells of EBV-associated Hodgkin's disease (Delsol et al, 1992; Pinkus et al, 1994; Oudejans et al 1997). With respect to antibodies against LMP, a note of caution is advised: strong staining of normal early myeloid and erythroid precursors may be seen despite a total absence of evidence of EBV by PCR (Hammer et al, 1996).
Comments
As a research tool, EBV immunohistochemical investigation is superior to PCR, in that the latter does not exclude background/resident lymphocytes harboring EBV.
References
•Bruin PCD 1993 Detection of Epstein-Barr virus nucleic acid sequences and protein in nodal T-cell lymphomas: relation between latent membrane protein-1 positivity and clinical course. Histopathology 23: 509-518.
•Delecluse H-J, Kremmer E, Rouault J-P et al 1995 The expression of Epstein-Barr virus latent proteins is related to the pathological feature of post-transplant lymphoproliferative disorders. American Journal of Pathology 146: 1113-1120.
•Delsol G, Brousset P, Chittal S, Rigal HF 1992. Correlation of the expression of Epstein-Barr virus latent membrane protein and in situ hybridization with biotinylated BamH1-W probes in Hodgkin's disease. American Journal of Pathology 140:247-253.
•Dimery IW, Lee JS, Blick M et al 1988. Association of the Epstein-Barr virus with lymphoepithelioma of the thymus. Cancer 61: 2475-2480.
•Hamilton-Dutoit SJ, Rea D, Raphael M et al 1993. Epstein-Barr virus-latent gene expression and tumor cell phenotype in acquired immunodeficiency syndrome-related non-Hodgkin's lymphoma: correlation of lymphoma phenotype with three distinct patterns of viral latency. American Journal of Pathology 143: 1072-1090.
•Hammer RD, Scott M, Shahab I et al 1996. Latent membrane protein antibody reacts with normal haematopoietic precursor cells and leukaemic blasts in tissues lacking EBV by PCR. American Journal of Clinical Pathology 106: 469-474.
•Herbst H, Dallenbach F, Hummel M, et al 1991. Epstein-Barr virus DNA and latent gene products in Ki-1 (CD30)-positive anaplastic large cell lymphomas. Blood 78: 2663-1673.
•Hording U, Nielsen HW, Albeck H, Daugaard S 1993 Nasopharyngeal carcinoma: histopathological types and association with Epstein-Barr virus. European Journal of Cancer and Clinical Oncology 29B: 137-139.
•Kaczorowski S, Kaczorowska M, Christenson B 1994 Expression of EBV encoded latent membrane protein 1 and bcl-2 protein in childhood and adult Hodgkin's disease: application of microwave irradiation for antigen retrieval. Leukemia and Lymphoma 13: 273-283.
•Kanavaros P, Lecsc M-C, Briere J et al 1993. Nasal T-cell lymphoma: a clinicopathologic entity associated with peculiar phenotype and with Epstein-Barr virus. Blood 81: 2688-2695.
•Lopategui JR, Gaffey MJ, Frierson HF et al 1994. Detection of Epstein-Barr viral RNA in sinonasal undifferentiated carcinoma from Western and Asian patients. American Journal of Surgical Pathology 18:391-398.
•Oudejans JJ, Jiwa NM, Meijer CJLM 1997 Epstein-Barr virus in Hodgkin's disease: more than just an innocent bystander. Journal of Pathology 181: 353-356
•Pinkus GS, Lones M, Shinataku IP, Said JW 1994. Immunohistochemical detection of Epstein-Barr virus-encoded latent membrane protein in Reed-Sternberg cells and variants of Hodgkin's disease. Modern Pathology 7: 454-461.
•Rowe M, Evans HS, Young LS et al 1989. Monoclonal antibodies to the latent membrane protein of Epstein-Barr virus reveal heterogeneity of the protein and inducible expression in virus transformed cells. Journal of General Virology 68: 1575-1586.
•Shibata D, Tokunaga M, Uemura Y et al 1991. Association of EBV with undifferentiated gastric carcinomas with intense lymphoid infiltration. Americal Journal of Pathology 139: 469-474.
•Tokunaga M, Imai S, Utemura Y, Tokudome T, Osato T, Sato E 1993 Epstein-Barr virus in adult T-cell leukaemia/lymphoma. American Journal of Pathology: 1263-1269.
•Weiss LM, Movahed LA, Butler AE et al 1989. Analysis of lymphoepithelioma and lymphoepithelioma-like carcinoma for Epstein-Barr viral genomes by in situ hybridization. American Journal of Surgical Pathology 13: 625-631.
Bibliografía
Manual of diagnostic antibodies for immunohistology / Anthony S.-Y. Leong, Kumarasen Cooper, F. Joel W.-M. Leong.
Accurate (CS1-4), Biodesign (polyclonal), Dako (CS1-4), EY Labs, Novocastra (polyclonal) and Research Diagnostics (4C5).
Fixation/Preparation
Applicable to formalin-fixed, paraffin-embedded tissue sections. Enzymatic digestion (e.g. trypsin) is essential to enhance immunopositivity. The application of microwave irradiation for antigen retrieval has also been used to good effect (Kaczorowski et al, 1994). This antibody may also be used for labeling acetone-fixed cryostat sections or fixed-cell smears.
Background
The antibody (isotype: IgG1, k) has been raised against recombinant fusion protein containing sequences of bacterialb -galactosidase and the EBV-encoded latent membrane protein (LMP-1). LMP is one of the few viral proteins that are expressed in a latent infection. The antibody reacts with a 60 kD latent membrane protein encoded by the BNLF gene of the Epstein-Barr virus. Being a cocktail of clones CS1, CS2, CS3 and CS4, all four anti-LMP antibodies recognize distinct epitopes on the hydrophilic carboxyl region of LMP (Rowe et al, 1987). These four epitopes are present on the internal aspect of the membrane-associated viral LMP. Therefore the antibody does not react with viable cells, but with fixed cells in paraffin sections, cytological preparations and cryostat sections and in immunoblotting.
Applications
The antibody is characterized by its strong positivity with EBV+ lymphoblastoid cell lines and EBV-infected B-cell immunoblasts in infectious mononucleosis. Although EBV is consistently present in nasopharyngeal undifferentiated carcinoma among Oriental patients, LMP-1 antibody is only positive in about 60% of cases (Hording et al, 1993; Lopategui et al, 1994). LMP protein expression is especially useful in identifying these cancers in cervical lymph node metastases. This antibody may also be useful in the diagnosis of lymphoepithelioma-like carcinoma of the lung, mediastinum, stomach and paranasal sinuses (Dimery et al, 1988; Weiss et al, 1989; Shibata et al, 1991).
Posttransplantation lymphoproliferative disorders arising in patients treated with a variety of immunosuppressive regimens after organ transplantation usually show a type III latency pattern with LMP-1 expression (Delecluse et al, 1995). The EBV+ AIDS-associated B-cell lymphomas usually demonstrate a latency type III in the large cell lymphomas, permitting the use of antibody to LMP-1 (Hamilton-Du Toit et al, 1993).
Nasal T/NK-cell lymphoma is strongly associated with EBV (Kanavaros et al, 1993). However, LMP-1 protein expression has been inconsistent on paraffin sections, although one study consistently demonstrated LMP-1 protein in frozen sections, suggesting a low level of protein expression. LMP-1 immunohistochemistry is positive in 17% of adult T-cell leukemia/lymphoma (Tokunaga et al, 1993). LMP-1 expression has also been associated with an aggressive clinical course and hepatosplenomegaly in nodal T-cell lymphomas (Bruin, 1993). About 20-30% of CD 30 (Ki-1)+ anaplastic large cell lymphoma show LMP-1 immunoreaction (Herbst et al, 1991).
Approximately 50% of Hodgkin's disease cases are associated with EBV. In almost all of these positive cases, nearly all the Reed-Sternberg cells are positive for EBV. Using modern epitope retrieval techniques, an almost 1:1 correlation between the results of LMP-1 paraffin-based immunohistochemistry studies and EBER in situ hybridization studies has been demonstrated in Reed-Sternberg cells and Hodgkin's cells of EBV-associated Hodgkin's disease (Delsol et al, 1992; Pinkus et al, 1994; Oudejans et al 1997). With respect to antibodies against LMP, a note of caution is advised: strong staining of normal early myeloid and erythroid precursors may be seen despite a total absence of evidence of EBV by PCR (Hammer et al, 1996).
Comments
As a research tool, EBV immunohistochemical investigation is superior to PCR, in that the latter does not exclude background/resident lymphocytes harboring EBV.
References
•Bruin PCD 1993 Detection of Epstein-Barr virus nucleic acid sequences and protein in nodal T-cell lymphomas: relation between latent membrane protein-1 positivity and clinical course. Histopathology 23: 509-518.
•Delecluse H-J, Kremmer E, Rouault J-P et al 1995 The expression of Epstein-Barr virus latent proteins is related to the pathological feature of post-transplant lymphoproliferative disorders. American Journal of Pathology 146: 1113-1120.
•Delsol G, Brousset P, Chittal S, Rigal HF 1992. Correlation of the expression of Epstein-Barr virus latent membrane protein and in situ hybridization with biotinylated BamH1-W probes in Hodgkin's disease. American Journal of Pathology 140:247-253.
•Dimery IW, Lee JS, Blick M et al 1988. Association of the Epstein-Barr virus with lymphoepithelioma of the thymus. Cancer 61: 2475-2480.
•Hamilton-Dutoit SJ, Rea D, Raphael M et al 1993. Epstein-Barr virus-latent gene expression and tumor cell phenotype in acquired immunodeficiency syndrome-related non-Hodgkin's lymphoma: correlation of lymphoma phenotype with three distinct patterns of viral latency. American Journal of Pathology 143: 1072-1090.
•Hammer RD, Scott M, Shahab I et al 1996. Latent membrane protein antibody reacts with normal haematopoietic precursor cells and leukaemic blasts in tissues lacking EBV by PCR. American Journal of Clinical Pathology 106: 469-474.
•Herbst H, Dallenbach F, Hummel M, et al 1991. Epstein-Barr virus DNA and latent gene products in Ki-1 (CD30)-positive anaplastic large cell lymphomas. Blood 78: 2663-1673.
•Hording U, Nielsen HW, Albeck H, Daugaard S 1993 Nasopharyngeal carcinoma: histopathological types and association with Epstein-Barr virus. European Journal of Cancer and Clinical Oncology 29B: 137-139.
•Kaczorowski S, Kaczorowska M, Christenson B 1994 Expression of EBV encoded latent membrane protein 1 and bcl-2 protein in childhood and adult Hodgkin's disease: application of microwave irradiation for antigen retrieval. Leukemia and Lymphoma 13: 273-283.
•Kanavaros P, Lecsc M-C, Briere J et al 1993. Nasal T-cell lymphoma: a clinicopathologic entity associated with peculiar phenotype and with Epstein-Barr virus. Blood 81: 2688-2695.
•Lopategui JR, Gaffey MJ, Frierson HF et al 1994. Detection of Epstein-Barr viral RNA in sinonasal undifferentiated carcinoma from Western and Asian patients. American Journal of Surgical Pathology 18:391-398.
•Oudejans JJ, Jiwa NM, Meijer CJLM 1997 Epstein-Barr virus in Hodgkin's disease: more than just an innocent bystander. Journal of Pathology 181: 353-356
•Pinkus GS, Lones M, Shinataku IP, Said JW 1994. Immunohistochemical detection of Epstein-Barr virus-encoded latent membrane protein in Reed-Sternberg cells and variants of Hodgkin's disease. Modern Pathology 7: 454-461.
•Rowe M, Evans HS, Young LS et al 1989. Monoclonal antibodies to the latent membrane protein of Epstein-Barr virus reveal heterogeneity of the protein and inducible expression in virus transformed cells. Journal of General Virology 68: 1575-1586.
•Shibata D, Tokunaga M, Uemura Y et al 1991. Association of EBV with undifferentiated gastric carcinomas with intense lymphoid infiltration. Americal Journal of Pathology 139: 469-474.
•Tokunaga M, Imai S, Utemura Y, Tokudome T, Osato T, Sato E 1993 Epstein-Barr virus in adult T-cell leukaemia/lymphoma. American Journal of Pathology: 1263-1269.
•Weiss LM, Movahed LA, Butler AE et al 1989. Analysis of lymphoepithelioma and lymphoepithelioma-like carcinoma for Epstein-Barr viral genomes by in situ hybridization. American Journal of Surgical Pathology 13: 625-631.
Bibliografía
Manual of diagnostic antibodies for immunohistology / Anthony S.-Y. Leong, Kumarasen Cooper, F. Joel W.-M. Leong.
