Surface markers on human B‐ and T‐lymphocytes. IX. Two‐color immunofluorescence studies on the association between EBV receptors and complement receptors on the surface of lymphoid cell lines

Yefenof, Eitan; Klein, George; Jondal, Mikael; Oldstone, Michael B. A.

doi:10.1002/ijc.2910170602

Cited by 128 publications

(51 citation statements)

References 18 publications

(6 reference statements)

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“…This interpretation is also consistent with the proposal by Yefenof et al (16), based on cocapping experiments, that the EB virus receptor is closely linked to the EAC receptor on the human B lymphocyte. However, other interpretations of our data should be mentioned.…”

supporting

confidence: 93%

Host-determined differences in expression of surface marker characteristics on human and simian lymphoblastoid cell lines transformed by Epstein-Barr virus.

Robinson¹,

Andiman²,

Henderson³

et al. 1977

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

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In an attempt to account for differences in the biologic behavior of Epstein-Barr virus in different primate species, we studied lymphocyte surface markers on primary and transformed cells. Among primary leukocytes, the distribution of cells with characteristics of bone-marrow-derived cells ( MATERIALS AND 'METHODS Cell Culture. Mononuclear leukocytes from human umbilical cord blood or from peripheral blood of cotton-top marmosets or woolly monkeys were isolated on a Ficoll-Hypaque gradient (11). The cells were transformed by EB virus and maintained in RPMI-1640 with 20% fetal calf serum as described previously (8).Leukocyte Markers. E Rosettes. Leukocytes (0.2 to 1.0 X 106) were mixed with an equal volume of a 1% suspension of sheep red blood cells (SRBC) that had been pretreated with neuraminidase (Vibrio cholera, Calbiochem) (25 units/ml), centrifuged at 200 X g for 10 min, and held at 40 for 15 min or at room temperature for 1 hr (12). The pellet was gently resuspended, and the number of E-rosette forming cells (E-RFC), or thymus-derived lymphocytes, were counted.EA(7S) Rosettes. Equal volumes of a 5% suspension of SRBC and a nonagglutinating dilution of 7S rabbit antibody to SRBC were incubated at 370 for 30 min. The sensitized SRBC [EA(7S)] were washed twice and adjusted to a 1% suspension in Hanks' balanced salt solution. Leukocytes (0.2 to 1 X 106) were incubated with an equal volume of 1% EA(7S) at 370 for 30 min and the EA(7S)-RFC were counted. Untreated SRBC incubated with leukocytes at 370 did not form rosettes.EAC Rosettes. Equal volumes of 5% sheep erythrocytes and a nonagglutinating dilution of 19S rabbit antibody to sheep erythrocytes (Cordis Labs) were incubated at 370 for 30 min, washed twice in Hanks' balanced salt solution, adjusted to a 5% suspension, and incubated for 30 min at 370 with an equal volume of mouse serum diluted 1:10 in gelatin-barbital buffer (13). The mixture was washed twice more and adjusted to a 1% suspension in the salt solution. The assay for EAC-RFC was exactly the same as that for EA(7S)-RFC.Assay for Phagocytes. The EAC reagent treated with 7S antibody to SRBC as in the preparation of EA(7S) is a potent 749

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supporting

confidence: 93%

Host-determined differences in expression of surface marker characteristics on human and simian lymphoblastoid cell lines transformed by Epstein-Barr virus.

Robinson¹,

Andiman²,

Henderson³

et al. 1977

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

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“…The majority of B lymphocytes that bind EBV are those with surface immunoglobulins (Anderson et al, 1983;Katsuki et al, 1977) and in resting phase (Go-Gag) (Wells et al, 1981). Interest in an EBV receptor (EBV-R) has grown since the discovery of an association between EBV-R and the C3d (complement) receptor (Freeman et al, 1982;Jondal et al, 1976;Jonsson et al, 1982;Yefenof et al, 1976Yefenof et al, , 1977. Better understanding of complement receptors (Fearon, 1985) and the availability of monoclonal antibodies (MAbs) directed against the C3d receptor (CR2) (Fingeroth et al, 1984;Iida et al, 1983;Nemerow et al, 1985;Slaw et al, 1986; has led to the demonstration of the association between EBV-R and the CR2 on B cells.…”

Section: Introductionmentioning

confidence: 99%

Epstein--Barr virus receptor expression on human CD8+ (cytotoxic/suppressor) T lymphocytes

Sauvageau

Stocco

Kasparian

et al. 1990

Journal of General Virology

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In 1977 we showed that cells of a human lymphocytic leukaemia-derived T line (Molt-4) have receptors for Epstein-Barr virus (EBV). More recently, EBVpositive human T cell lymphomas have been recognized and human T cell lines containing the EBV genome have been established in vitro. To understand better the interaction of EBV with T cells, we decided to determine first whether human peripheral blood T lymphocytes express receptors for EBV. Using flow cytometry we examined the binding of both lymphocyte-transforming (B95-8) and non-transforming (P3HR-1) strains of EBV to T lymphocyte subpopulations, using a double labelling technique with T cellspecific phycoerythrinated monoclonal antibodies (Leu 2a) and fluoresceinated viral preparation. Our results suggest that, in general, about 50% of the CD8 + (or suppressor/cytotoxic) T cell subpopulation from both EBV-seropositive and -seronegative individuals can bind EBV. EBV receptor expression on these T cells was about l0 and 51 times less than that on Molt-4 and Raji (an EBV receptor-positive B cell line) cells, respectively. The specificity of this binding was demonstrated by the inhibition of attachment of viral preparations preincubated with a monoclonal antibody directed against the viral ligand (gp240/350), and by preincubating these target T cells with unlabelled virus. We were unable to detect EBV-induced antigens in infected T cells, suggesting that, as in Molt-4 cells, virus internalization may not occur in fresh T cells and/or that the virus receptor may not be completely functional. We were also unable to detect C3d (or CR2) receptors on these T cells, or to inhibit virus attachment by treating the targets with an anti-CR2 monoclonal antibody (OKB7), suggesting that the EBV receptor on CD8 + peripheral blood lymphocytes is different from that on B cells.

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“…It may now be possible to delineate portions of the EBV genome needed for immortalization, provided that a virion-associated polymerase is not needed for the infectivity of EBV DNA. Another objective would be to learn whether the strict B cell tropism of EBV, which seems to be determined by an interaction between the virus and a specific receptor on B lymphocytes, can be overcome by using EBV DNA (15). It would then be possible to learn whether the highly efficient transforming capacities of the EBV genome can be expressed in other hematopoietic or tissue cell types.…”

Section: Susceptibilitymentioning

confidence: 99%

Transfection of human lymphoblastoid cells with herpes simplex viral DNA.

Miller¹,

Wertheim²,

Wilson³

et al. 1979

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

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The "calcium/dimethyl sulfoxide shock" method of transfection was adapted for use in human lymphoid cell cultures. One microgram of herpes simplex virus type 1 DNA regularly initiated virus replication in four lymphoblastoid cell lines. Per 105 cells exposed to 1 pg of DNA, 0.5-5 cells formed an infectious center. The minimal infective dose of DNA was approximately 500 ng.A reliable method for transfection of human lymphocytes by DNA could have many applications in the study of lymphotropic viruses and also might be useful in experimental immunology and genetics. To initiate studies of this problem we chose a system that would maximize the chances of demonstrating expression of genetic information after introduction of naked DNA into lymphoid cells. The DNA of herpes simplex virus (HSV) was selected because one assay for expressionnamely, production of complete progeny-is simple and rapid and because several methods are already available for detection of infectious herpes DNA on monolayer tissue culture cells (1-3). The ability to assay HSV-1 DNA independently in permissive cells provided a check on the quality of the DNA and on the adequacy of the transfection technique. HSV DNA was chosen also because it is a good model for future studies with Epstein-Barr virus (EBV) DNA, a molecule of the same size.MATERIALS AND METHODS Virus Strain, Propagation, and Preparation of Viral DNA. The Lovering strain of HSV-1, originally from a fetal encephalitis case, was passed at low multiplicity three times in the AH-1 and once in the BSC-1 line of African green monkey kidney cells (4). The virus was plaque-purified in BSC-1 cells under 2% methylcellulose. Two batches of viral DNA were used: batch 4, prepared from virions propagated in BSC-1 cells; batch 6, derived from a subclone obtained in transfection of BSC-1 cells, was prepared from virions propagated in BHK-21 cells.Viral DNA was obtained from virions by the procedure described by Geelen et al. (5) for preparation of infectious cytomegalovirus DNA. The starting material for each batch of DNA was 6-20 roller bottles of BSC-1 cells. The medium was removed from confluent cultures, which were inoculated with about 2 plaque-forming units (PFU) per cell. After 1 hr the inoculum was removed and 50 ml of Eagle's medium with 2% calf serum was added to each bottle. After 40 hr, when there was extensive cytopathic effect, the culture medium was decanted and centrifuged at 4000 rpm at 4°C for 10 min in the Sorvall GSA rotor. The supernatant fluids were separated from the pellets and centrifuged for 3 hr at 18,000 rpm Lymphoid Cells. Three lymphoblastoid lines, designated X25, were derived after transformation of lymphocytes from one umbilical cord with a passage of the B95-8 strain of EBV (unpublished data); the fourth line (X50-7) originated from another umbilical cord. These lines are not spontaneous producers of extracellular EBV but transforming virus can be recovered from the lines by the technique of x-irradiation and cocultivation. All the cells have EBV nuclear ...

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Surface markers on human B‐ and T‐lymphocytes. IX. Two‐color immunofluorescence studies on the association between EBV receptors and complement receptors on the surface of lymphoid cell lines

Cited by 128 publications

References 18 publications

Host-determined differences in expression of surface marker characteristics on human and simian lymphoblastoid cell lines transformed by Epstein-Barr virus.

Host-determined differences in expression of surface marker characteristics on human and simian lymphoblastoid cell lines transformed by Epstein-Barr virus.

Epstein--Barr virus receptor expression on human CD8+ (cytotoxic/suppressor) T lymphocytes

Transfection of human lymphoblastoid cells with herpes simplex viral DNA.

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