- Characteristics of the Immortalized Human B-cells by Epstein-Barr Virus.
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Ho Jong Jeon, Bong Nam Choi, Yoon Kyeong Oh
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Korean J Pathol. 1997;31(9):832-846.
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 Abstract
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- Human lymphoblastoid B-cell lines immortalized by Epstein-Barr virus (EBV) were established from peripheral blood of patients with acute myeloblastic and chronic lymphocytic leukemia and chronic fatigue syndrome. The sera of patients with acute myeloblastic and chronic lymphocytic leukemia did not show antibodies to Epstein-Barr viral capsid antigen (VCA), but serum of a patient with chronic fatigue syndrome disclosed antibodies to VCA (IgG, IgM), and EBNA was demonstrated in peripheral blood mononuclear cells by polymerase chain reaction. The established cell lines were mature B-cell phenotypes with polyclonal proliferation in early passage and no evidence for commitment to other lineages. The immortalized cells by EBV were designated as CSUP-1 and CSUP-2 (from acute myeloblastic leukemia, FAB classification M2 and M1), CSUP-3 (from chronic lymphocytic leukemia) and CSUP-4 (from a patient with chronic fatigue syndrome). The CSUP-1, 2, 3, and 4 grew in suspension forming clumps with a doubling time of 38 to 49 hours.
Colony formation was not recognized in plate. By light and electron microscopic examination, the immortalized cells showed features of lymphoblastoid to plasmacytoid lymphocytes, and multinucleated giant cells. The lymphoblastoid cells showed scanty cytoplasm with poorly developed organelles. Immunophenotypic analyses of CUSP-1, 2, 3, and 4 with monoclonal antibodies by flow cytometry showed B-cell phenotype with polyclonal proliferation in early passage. Epstein-Barr virus nuclear antigen was confirmed in the extracted DNAs from immortalized cells by polymerase chain reaction. DNA analysis showed a normodiploid stemline with a DNA index of 1.12. The established cells were strongly reactive for CD10, CD30 (Ki-1) in early passage, and bcl-2 and c-myc onco-protein in early and late passage. Karyotypic analysis of CSUP-1, 2, 3 and 4 showed 46, XY or 46, XX. No tumorigenesis in heterotransplanted SCID mouse was recognized. This immortalized cells by EBV should be a valuable cell lines to study the pathogenesis of EBV-related malignant lymphoma.
- An Immunohistochemical Study on the Expression of the Bauhinia Purpurea in the Reed-Sternberg Cells.
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Yun Sin Kim, Mi Sook Lee, Ho Jong Jeon, Bong Nam Choi, Jong Hoon Jung, Choon Hae Chung, Chul Woo Kim
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Korean J Pathol. 1995;29(4):459-468.
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Abstract
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- The diagnosis of Hodgkin's disease is based on the morphologic identification of Reed-Sternberg (RS) cells and its variants in paraffin-embedded sections. The origin of RS cells remains a subject of controversy, and cells resembling RS cells are observed in some non-Hodgkin's lymphoma of T-cell lineage. In this study, eighteen cases of Hodgkin's disease (3 nodular sclerosis, 6 diffuse lymphocyte predominance, and 9 mixed cellularity) were studied with peanut agglutinin(PNA), anti-Leu-M1(CD15), LN2(CD74), Ber-H2(CD30) and bauhinia purpurea (BPA) by the avidin-biotin-peroxidase complex(ABC) method in paraffin-embedded sections. RS cells and their variants revealed positive reactions with one or more of the reagents in all examined cases. BPA staining was positive in 17 of 18 cases (94.4%), PNA staining was positive in 9 of 18 cases (50.0%), Leu MI was positive in 7 of 18 cases(38.9%), Ber-H2 was positive in 11 of 18 cases (61.1%), and LN2 was positive in 8 of 18 cases(44.4%). The staining properties of examined markers were recognized as paranuclear, diffuse cytoplasmic and cellular membranous patterns, but LN2 disclosed diffuse cytoplasmic staining in the positive cells. BPA also showed dense cytoplasmic staining reaction with macrophage-histiocytes. BPA reactivity was not affected by fortnalin fixation or paraffm embedding. Thirty six cases of non-Hodgkin's lymphomas(IO T-cell and 26 B-cell type) were also examined. The neoplastic cells of those cases did not stain positive with BPA, PNA, and Leu-Mi, but stained positively with LN2 in 3 cases of T-cell lymphomas and 14 cases of B-cell lymphomas, and BeT-H2 in T-cell lymphomas.
In conclusion, to facilitate the detection of RS cells and related variants in paraffm sectionse of Hodgkin's disease, BPA can be used as a useful marker because of its high-detection rate, reproducible staining pattem, and resistance to fixative.
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