Surface Markers on Human B and T Lymphocytes

Jondal, Mikael; Klein, George

doi:10.1084/jem.138.6.1365

Cited by 494 publications

(121 citation statements)

References 19 publications

(19 reference statements)

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“…Epstein-Barr (EB) virus has the capacity to infect human bone marrow-derived (B-) lymphocytes (Jondal & Klein, 1973;Pattengale et al I973;Greaves et al I975) and to induce their transformation in culture to permanent lymphoblastoid lines (Henle et al 1967;Pope et al I968) in which essentially every cell carries multiple circular copies of the EB virus genome in some stable but non-integrated association with cellular DNA (zur Hausen et al 1972;Lindahl et al 1976), and expresses the virus-associated nuclear antigen (EBNA; Reedman & Klein, 1973). The precise sequence of events which must occur in an infected cell before the establishment of the transformed state is little understood, although it must clearly include the induction of EBNA and probably also the generation of multiple copies of the incoming virus genome by some process of amplification.…”

Section: Introductionmentioning

confidence: 99%

Sensitivity of the Transforming and Replicative Functions of Epstein-Barr Virus to Inhibition by Phosphonoacetate

Rickinson

Epstein

1978

Journal of General Virology

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SUMMARYDisodium, phosphonoacetate (PA), at concentrations of 50 to 2o0/~g/ml, which still allowed continued growth of the EB virus-transformed B95-8 cell line on a routine culture regimen, was able to inhibit the production of virus capsid antigen and of virus particles by these cells down to very low but finite levels which persisted despite prolonged treatment. Further experiments measured the effects of these same drug concentrations on the EB virus-induced in vitro transformation of foetal cord blood lymphocytes and on the colony forming ability of already established EB virus-transformed foetal cell lines; in both types of culture, doses of PA up to and including 5o #g/ml did not affect cell growth within the 8-week observation period, whereas doses of ioo #g/ml and above were increasingly inhibitory. The cell lines established by EB virus-induced transformation in the continual presence of PA at 50 to I5O #g/ml contained multiple copies of the virus genome per cell just as did the corresponding cell lines established in control medium. The results argue against the existence of any PA-sensitive event unique to the EB virus-induced transformation process.

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Section: Introductionmentioning

confidence: 99%

Sensitivity of the Transforming and Replicative Functions of Epstein-Barr Virus to Inhibition by Phosphonoacetate

Rickinson

Epstein

1978

Journal of General Virology

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“…We have chosen to investigate membrane receptors for Epstein-Barr virus (EBV-R) because of our interest in the host cell range and biological activity of this human tumor agent. EBV-R have been demonstrated only on human and certain other primate B lymphocytes (6,181, strengthening the association between the virus and Burkitt's lymphoma (9). However, viral genome and nuclear antigen, EBNA, have also been regularly found in nasopharyngeal carcinoma cells (8,131.…”

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confidence: 99%

Application of flow cytometry to studies on the distribution, persistence, and regeneration rate of Epstein‐Barr virus receptors

McCune

Volsky

1984

Cytometry

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Flow cytometry has been applied to study the persistence and regeneration rate of Epstein-Barr virus (EBV) receptors and other membrane proteins in normal and tumor cells. EBV receptors were detected in a binding assay utilizing fluorescein isothiocyanate-conjugated virions (FITC-EBV). After stripping receptors from the cell surface with trypsin, binding of FITC-EBV was rapidly regenerated in the lymphoma cell line Loukes, reaching 75% of the control levels by 3 h. Fresh human lymphocytes regenerated receptors at a much slower rate, reaching 50% of control levels by 24 h. The regenerating receptors did not reappear simultaneously on all the cells. Subpopulation of new receptor-positive cells increased gradually during the incubation at 37°C. Reappearance of receptors was inhibited in the absence of protein synthesis. Conversely, receptors persisted in the nontrypsinized cells in the absence of protein synthesis. These results show that certain parameters of membrane receptor turnover, such as the change in receptor distribution within a cell population, receptor persistance, and regeneration rate, can be measured at the single-cell level by flow cytometry.Key terms: Flow cytometry, Epstein-Barr virus (EBV), surface receptors, membrane protein turnover, fluoresceinated virus Recent approaches to the study of membrane protein turnover have involved detecting specific proteins by the time-consuming techniques of sodium dodecyl sulfate (SDS)-gel electrophoresis (5), immunoprecipitation (l), purification of individual proteins (201, and the use of fluorescent probes and fluorescence microscopy (2). Only the latter technique permits following the fate of a n individul membrane protein on single cells, but it is qualitative and insensitive. Flow cytometry offers a rapid, sensitive, and quantitative method of detecting cell surface markers at the single cell level (10). Here we describe flow cytometry as a convenient and effective tool for the study of two aspects of membrane protein turnover: the persistence of membrane receptors in the presence and absence of protein synthesis inhibitor and regeneration of membrane receptors after stripping with trypsin. We have chosen to investigate membrane receptors for Epstein-Barr virus (EBV-R) because of our interest in the host cell range and biological activity of this human tumor agent. EBV-R have been demonstrated only on human and certain other primate B lymphocytes (6,181, strengthening the association between the virus and Burkitt's lymphoma (9). However, viral genome and nuclear antigen, EBNA, have also been regularly found in nasopharyngeal carcinoma cells (8,131. This indicates that under certain pathologic conditions EBV can bind to and penetrate into human epithelial cells. Moreover, while EBV-infected lymphocytes are normally recognized and eliminated by the immune system (41, the BL and NPC cells can successfully avert immune response (14), presumably as a result of some unknown malignancy-related cell surface changes. We have compared the persistence and regenerat...

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“…Most human lymphoid cell lines carrying the Epstein-Barr (EB) virus genome have characteristics of bone-marrow-derived (B) lymphocytes (i.e., surface immunoglobulin or receptors for fixed complement) (1). Therefore, it has been postulated that EB virus transforms only the B subpopulation of human lymphoid cells.…”

mentioning

confidence: 99%

“…Removal of B cells decreases the efficiency of transformation, and enrichment of B cells enhances it (4,5). Only B cells form rosettes with the cytoplasmic membrane of the EB virus-producer human lymphoid cell line, P3J-HR-1 (1). This finding suggests that only B cells have receptors for EB virus, an idea that is supported by the demonstration that supernatant fluids of two EB virus producer cell lines contain materials (presumably virions and virus altered cell membranes) which adsorb selectively to B cells (6,7).…”

mentioning

confidence: 99%

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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Surface Markers on Human B and T Lymphocytes

Cited by 494 publications

References 19 publications

Sensitivity of the Transforming and Replicative Functions of Epstein-Barr Virus to Inhibition by Phosphonoacetate

Sensitivity of the Transforming and Replicative Functions of Epstein-Barr Virus to Inhibition by Phosphonoacetate

Application of flow cytometry to studies on the distribution, persistence, and regeneration rate of Epstein‐Barr virus receptors

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

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