Radioimmune precipitation study comparing the epstein-barr virus membrane antigens expressed on P3HR-1 virus-superinfected Raji cells to those expressed on cells in a B-95 virus-transformed producer culture activated with tumor-promoting agent (TPA)

Qualtiere, Louis F.; Pearson, Gary R.

doi:10.1016/0042-6822(80)90103-8

Cited by 45 publications

(37 citation statements)

References 21 publications

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“…For that reason the resulting mRNAs from the CHO clones are about 130 nt larger in size. The distribution of the two transcripts reflects the situation found on the protein level: the CHO clones produce almost the same ratio, B95-8 cells mainly the gp350 variant and p3HRl cells only the gp250 (Qualtiere and Pearson, 1980). The amplification of the inserted sequences in the CHO clones with methotrexate and the purification of the gp250/350 are in progress.…”

Section: Kb Kbmentioning

confidence: 65%

Expression of the Epstein-Barr virus major membrane proteins in Chinese hamster ovary cells

Motz¹,

Deby²,

Jilg³

et al. 1986

Gene

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SUMMARYThe gene of the major membrane antigen (gp250/350) of the Epstein-Barr virus (EBV) was isolated and inserted under the control of the SV40 early promoter into a eukaryotic expression vector, which allows selection for dhfr' phenotype. Following transfection with this vector, Chinese hamster ovary cells express on their surfaces proteins immunologically similar to the major EBV membrane antigen. The transcript encoding gp250/350 is processed by partial splicing similarly but more efficiently than in B95-8 cells from which the DNA originates.

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Section: Kb Kbmentioning

confidence: 65%

Expression of the Epstein-Barr virus major membrane proteins in Chinese hamster ovary cells

Motz¹,

Deby²,

Jilg³

et al. 1986

Gene

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“…Analysis of extracellular virus particles produced by these two cell lines failed to reveal any differences in the major polypeptides present in them (13). More recently, however, analysis of EBV-induced membrane glycoproteins has revealed differences between cells infected with the P3HR-1 and B-95 viruses (14). Whether these differences were true strain differences or species differences due to the fact that the viruses were cultivated in cells from different species, as suggested by other authors (15), was unclear.…”

Section: Differencesmentioning

confidence: 86%

“…The cell lines used in this study included the virus-producing P3HR-1 cell line, a B-95 virus-induced cottontop marmoset tumor cell line (16) (14). Such (14,19) were mixed with 1% Triton X-100 in phosphate-buffered saline (PJ NaCl) containing 2 mM phenylmethylsulfonyl fluoride (Pierce) at a concentration of 1 ml of solvent per 2 x 107 cells. After 30 min at 25°C, nuclei and particulate matter were removed by centrifugation at 10,000 x g for 1 hr.…”

Section: Methodsmentioning

confidence: 99%

“…The mixtures were then incubated overnight at 40C. Immune complexes were removed by precipitation with protein A-Sepharose 4B (Pharmacia) as outlined in detail elsewhere (14,19). The complexes were then analyzed for the presence of EBV-specific membrane components by polyacrylamide gel electrophoresis as described (14,19).…”

Section: Methodsmentioning

confidence: 99%

“…Immune complexes were removed by precipitation with protein A-Sepharose 4B (Pharmacia) as outlined in detail elsewhere (14,19). The complexes were then analyzed for the presence of EBV-specific membrane components by polyacrylamide gel electrophoresis as described (14,19). The 14C-labeled proteins used as molecular weight standards were myosin (210,000), phosphorylase b (92,500), human serum albumin (69,000), ovalbumin (46,000), carbonic anhydrase (30,000), and lactoglobulin A (18,310) (New England Nuclear).…”

Section: Methodsmentioning

confidence: 99%

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Identification of Epstein-Barr virus strain differences with monoclonal antibody to a membrane glycoprotein.

Qualtiere¹,

Chase²,

Vroman³

et al. 1982

Proc. Natl. Acad. Sci. U.S.A.

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Two monoclonal antibodies directed against Epstein-Barr virus (EBV)-induced membrane antigens (MA) were isolated in this study. One of the monoclonal antibodies, designated 2F5.6, was an IgG2 which, as detected by membrane and fixed cell immunofluorescence, reacted with MA-positive lymphoblastoid cell lines that produced transforming EBV but not with the MA-positive P3HR-1 cell line that produced the lytic, nontransforming strain of this virus. This antibody precipitated the Mr 320,000/350,000 glycoprotein from B-95 virus infected cultures and the Mr 300,000 and 220,000/250,000 glycoproteins from Raji cells superinfected with P3HR-1 virus but did not precipitate any ofthese EBV-specific glycoproteins from the P3HR-1 cell line. In contrast, the second Imonoclonal antibody, IgM designated B10.3, reacted with all virus-producing cell lines including the P3HR-1 cell line. The identity of the glycoprotein that serves as the target for this antibody is still unknown. Neither antibody had neutralizing activity against the B-95 or P3HR-1. strain of EBV. These results indicated that the 2F5.6 monoclonal antibody was directed against an antigenic determinant on the major membrane glycoprotein which is common to transforming strains of EBV but absent from the lytic P3HR-1 strain whereas the B10.3 monoclonal antibody was directed against a group-specific EBV-induced membrane determinant. The Epstein-Barr virus (EBV) has been studied extensively because of its possible involvement in the etiology of nasopharyngeal carcinoma and African Burkitt lymphoma. Such studies have led to the identification of EBV isolates with different biological activities (1). The two prototype viruses that have been utilized extensively in these investigations are the P3HR-1 virus produced by a subline derived from a cell line established from an African Burkitt lymphoma biopsy sample (Jijoye) (2) and the B-95 virus isolated from a patient with infectious mononucleosis and propagated in marmoset cells (3). Although the viruses produced by both of these cell lines are morphologically similar and can be neutralized by sera from EBV-infected individuals, the biological activity and biochemical composition of these viruses show some significant, differences.The P3HR-1 virus is cytolytic for B lymphocytes, the target cell for EBV, and has apparently lost its ability to immortalize or transform B lymphocytes (4, 5). Therefore, it is categorized as a lytic nontransforming virus. This virus does, not induce the expression of the EBV-induced nuclear antigen (EBNA) upon infection ofB lymphocytes nor does it stimulate DNA synthesis, two. characteristics of transforming strains of EBV (6). Upon superinfection of EBV-genome-positive nonproducer cell lines with P3HR-1 virus, the synthesis ofthe early antigen (EA) complex is initiated followed by a shutdown ofcell synthesis of macromolecules. and the eventual death of the infected cell (7,8).In contrast, transforming strains of EBV, including the B-95 virus, generally do not initiate synthesis of the EA...

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Restrictions upon epstein‐barr virus infection of the leukemic cell are demonstrated in patients with hairy cell leukemia

Sairenji

Reisert

Spiro

et al. 1983

Hematological Oncology

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SUMMARYWe tested the hypothesis that Epstein-Barr virus (EBV) might actually infect leukemic hairy cells in vivo by examining those cells for the EBV-receptor, EBV nuclear antigen (EBNA) and membrane antigen (MA), for spontaneous transformation and rescue of infectious virus and for presence of EBV genome. EBV-receptors were found on subpopulations of leukemic cells from each of 7 patients with hairy cell leukemia (HCL) tested. MA was present on low numbers (1-5 per cent) of fresh leukemic cells of 7 patients and in some instances occurred with a greater frequency after 3 to 5 days in culture, with or without 12-0-tetradecanoylphorbol-13-acetate. In 1 1 fresh leukemic cell preparations from 8 HCL patients, no EBNA was demonstrated. However, 2 samples after 4 days in culture expressed low frequencies of EBNA-positive cells. Spontaneous, EBV-positive cell lines were established with a high transformation efficiency from 3 HCL blood samples but not from 8 other specimens. Infectious EBV could be rescued from some hairy leukemic cell preparations by co-cultivation with cord blood lymphocytes. These results demonstrated that leukemic cell populations harbored infectious EBV. that the leukemic cells expressed virus receptors and suggested that a small subpopulation of leukemic cells might become infected in viiw at least transiently and possibly transformed in vitro by EBV. To test for the extent of occult in vivo infection of leukemic cells with EBV, Southern type hybridization studies were performed with a probe for EBV genome (Barn HI W). At a sensitivity level of 0.1 genome per cell, EBV genome was not detected in the leukemic cell populations of 7 patients. We conclude that host defence mechanisms protecting these individuals from EBV also prevent infections of the leukemic cell and/or most hairy leukemic cells are not suitable targets for both infection and transformation.

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Radioimmune precipitation study comparing the epstein-barr virus membrane antigens expressed on P3HR-1 virus-superinfected Raji cells to those expressed on cells in a B-95 virus-transformed producer culture activated with tumor-promoting agent (TPA)

Cited by 45 publications

References 21 publications

Expression of the Epstein-Barr virus major membrane proteins in Chinese hamster ovary cells

Expression of the Epstein-Barr virus major membrane proteins in Chinese hamster ovary cells

Identification of Epstein-Barr virus strain differences with monoclonal antibody to a membrane glycoprotein.

Restrictions upon epstein‐barr virus infection of the leukemic cell are demonstrated in patients with hairy cell leukemia

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