Human Epstein-Barr virus (EBV)-specific cytotoxic T lymphocytes home preferentially to and induce selective regressions of autologous EBV-induced B cell lymphoproliferations in xenografted C.B-17 scid/scid mice.

C.B-17 scid/scid (severe combined immunodeficiency [SCID]) mice inoculated with peripheral blood lymphocytes from Epstein-Barr virus (EBV)-seropositive donors, or with EBV-transformed lymphoblastoid B cell lines (EBV-LCL), develop lethal human EBV+ B cell lymphoproliferative disorders (EBV-LPD) with characteristics similar to those arising in immunodeficient patients. Using this model, we examined the capacity of human effector cells to control human EBV-LPD. SCID mice received rabbit anti-asialo GM1 antiserum to abrogate endogenous natural killer-cell function. Preliminary experiments showed that adoptive transfer of peripheral blood mononuclear cells (PBMC), purified T cells, interleukin (IL) 2-activated PBMC or anti-CD3-activated T cells derived from EBV-seropositive donors did not result in improved survival of treated mice (in vivo effector/target ratio 2:1 to 1:1). In contrast, EBV-specific cytotoxic T lymphocytes (CTL), derived from EBV-seropositive donors and expanded in vitro, exhibited strong EBV-specific and HLA-restricted activity both in vitro and in vivo. SCID mice inoculated intraperitoneally with autologous but not with HLA-mismatched EBV-LCL had significantly improved survival relative to untreated mice after inoculation of EBV-specific CTL either intraperitoneally (P<0.001) or intravenously (P<0.001) (in vivo effector/target ratio 1:1). SCID mice bearing large subcutaneous EBV+ tumors and treated intravenously with 10(7) EBV-specific CTL achieved complete tumor regression. Both CTL- and CTL-plus-IL-2-treated mice survived significantly longer than untreated animals or animals treated with IL-2 alone (P = 0.0004 and P<0.02, respectively). SCID mice bearing two subcutaneous EBV+ tumors, one autologous and the other HLA mismatched to the EBV-specific CTL donor, had regression of only the autologous tumor after intravenous infusion of 10(7) EBV-specific CTL. Moreover, we could demonstrate preferential homing of PKH26-labeled EBV-specific CTL to autologous but not to HLA-mismatched EBV+ tumors as early as 24 h after intravenous adoptive transfer. Immunophenotypic analyses also demonstrated preferential infiltration of T cells into the autologous EBV+ tumor in SCID mice bearing both the autologous and either fully HLA-mismatched or genotypically related haplotype-sharing EBV+ tumors. The human T cells infiltrating EBV+ tumors were CD3+ and, predominantly, CD8+CD4-. Our results indicate that EBV-specific CTL preferentially localize to and infiltrate EBV+ tumors bearing the appropriate HLA antigens and thereafter induce targeted regressions of disease.

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Epstein-Barr virus, B cell lymphoproliferative disease, and SCID mice: Modeling T cell immunotherapy in vivo

Journal of Medical Virology ◽

10.1002/jmv.22164 ◽

2011 ◽

Vol 83 (9) ◽

pp. 1585-1596 ◽

Cited By ~ 7

Author(s):

I. Johannessen ◽

L. Bieleski ◽

G. Urquhart ◽

S.L. Watson ◽

P. Wingate ◽

...

Keyword(s):

T Cell ◽

B Cell ◽

Epstein Barr Virus ◽

Lymphoproliferative Disease ◽

Scid Mice ◽

Barr Virus ◽

Cell Immunotherapy ◽

Epstein Barr ◽

T Cell Immunotherapy

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Epstein-Barr Virus LMP2A Alters In Vivo and In Vitro Models of B-Cell Anergy, but Not Deletion, in Response to Autoantigen

Journal of Virology ◽

10.1128/jvi.79.12.7355-7362.2005 ◽

2005 ◽

Vol 79 (12) ◽

pp. 7355-7362 ◽

Cited By ~ 39

Author(s):

Michelle A. Swanson-Mungerson ◽

Robert G. Caldwell ◽

Rebecca Bultema ◽

Richard Longnecker

Keyword(s):

B Cells ◽

B Cell ◽

Epstein Barr Virus ◽

Nuclear Translocation ◽

Cell Receptor ◽

Barr Virus ◽

Low Levels ◽

Epstein Barr

ABSTRACT A significant percentage of the population latently harbors Epstein-Barr virus (EBV) in B cells. One EBV-encoded protein, latent membrane protein 2A (LMP2A), is expressed in tissue culture models of EBV latent infection, in human infections, and in many of the EBV-associated proliferative disorders. LMP2A constitutively activates proteins involved in the B-cell receptor (BCR) signal transduction cascade and inhibits the antigen-induced activation of these proteins. In the present study, we investigated whether LMP2A alters B-cell receptor signaling in primary B cells in vivo and in vitro. LMP2A does not inhibit antigen-induced tolerance in response to strong stimuli in an in vivo tolerance model in which B cells are reactive to self-antigen. In contrast, LMP2A bypasses anergy induction in response to low levels of soluble hen egg lysozyme (HEL) both in vivo and in vitro as determined by the ability of LMP2A-expressing HEL-specific B cells to proliferate and induce NF-κB nuclear translocation after exposure to low levels of antigen. Furthermore, LMP2A induces NF-κB nuclear translocation independent of BCR cross-linking. Since NF-κB is required to bypass tolerance induction, this LMP2A-dependent NF-κB activation may complete the tolerogenic signal induced by low levels of soluble HEL. Overall, the findings suggest that LMP2A may not inhibit BCR-induced signals under all conditions as previously suggested by studies with EBV immortalized B cells.

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Analysis of the defects responsible for the impaired regulation of Epstein-Barr virus-induced B cell proliferation by rheumatoid arthritis lymphocytes. I. Diminished gamma interferon production in response to autologous stimulation.

Journal of Experimental Medicine ◽

10.1084/jem.157.1.173 ◽

1983 ◽

Vol 157 (1) ◽

pp. 173-188 ◽

Cited By ~ 101

Author(s):

F Hasler ◽

H G Bluestein ◽

N J Zvaifler ◽

L B Epstein

Keyword(s):

Cell Proliferation ◽

T Cells ◽

T Cell ◽

B Cell ◽

Epstein Barr Virus ◽

Gamma Interferon ◽

Barr Virus ◽

T Cell Regulation ◽

Epstein Barr

T cells of patients with rheumatoid arthritis (RA) do not control the rate of B lymphoblast transformation induced by Epstein-Barr virus (EBV) as efficiently as T cells from healthy individuals; thus, lymphoblast cell lines are established more readily in RA lymphocytes in vitro after EBV infection. In the present experiments, we have asked whether this T cell regulation can be reproduced by lymphocytes. We found that normal T cells, activated in allogeneic or autologous mixed leukocyte reactions (MLR), produce lymphokines that inhibit in vitro EBV-induced B cell proliferation. Allogeneic MLR supernatants inhibited EBV-induced DNA synthesis 62 +/- 4% (mean +/- SE) at 10 d post-infection, whereas autologous MLR supernatants suppressed it 50 +/- 3%. RA T cell supernatants produced in an allogeneic MLR suppressed as well as normal T cell supernatants (64 +/- 5% inhibition). In contrast, supernatants from RA autologous MLR had little inhibitory activity. EBV-induced DNA synthesis at 10 d was reduced only 8 +/- 3%, compared with the 50 +/- 3% suppressive activity of normal autologous MLR supernatants. The magnitude of the proliferative responses in the autologous MLR regenerating the lymphokines was similar in the normal and RA populations. After depletion of adherent cells from the RA auto-MLR stimulators, supernatant inhibitory activities increased to normal levels (from 11 +/- 6 [SE] to 52 +/- 6% [SE]). The inhibitory factor involved in the regulation of in vitro EBV infection is a protein with a molecular weight of approximately 50,000. Its activity is eliminated by hearing at 56 degrees C and by exposure to acid at pH 2. The inhibitory activity is blocked by mixing the MLR supernatants with a polyvalent antisera or monoclonal antibodies specific for human gamma interferon. Gamma interferon produced by activating T cells in allo- or auto-MLR can reproduce T cell-mediated regulation of EBV-induced B cell proliferation, and the failure of RA auto-MLR to generate that lymphokine parallels the defective T cell regulation of EBV-induced B cell proliferation characteristic of RA lymphoid cells.

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Molecular virology of Epstein–Barr virus

Philosophical Transactions of the Royal Society B Biological Sciences ◽

10.1098/rstb.2000.0781 ◽

2001 ◽

Vol 356 (1408) ◽

pp. 437-459 ◽

Cited By ~ 99

Author(s):

Georg W. Bornkamm ◽

Wolfgang Hammerschmidt

Keyword(s):

B Cell ◽

Epstein Barr Virus ◽

Descriptive Analysis ◽

Genetic System ◽

Lytic Cycle ◽

Barr Virus ◽

Cell Immortalization ◽

Epstein Barr

Epstein–Barr virus (EBV) interacts with its host in three distinct ways in a highly regulated fashion: (i) EBV infects human B lymphocytes and induces proliferation of the infected cells, (ii) it enters into a latent phase in vivo that follows the proliferative phase, and (iii) it can be reactivated giving rise to the production of infectious progeny for reinfection of cells of the same type or transmission of the virus to another individual. In healthy people, these processes take place simultaneously in different anatomical and functional compartments and are linked to each other in a highly dynamic steady–state equilibrium. The development of a genetic system has paved the way for the dissection of those processes at a molecular level that can be studied in vitro , i.e. B–cell immortalization and the lytic cycle leading to production of infectious progeny. Polymerase chain reaction analyses coupled to fluorescent–activated cell sorting has on the other hand allowed a descriptive analysis of the virus–host interaction in peripheral blood cells as well as in tonsillar B cells in vivo . This paper is aimed at compiling our present knowledge on the process of B–cell immortalization in vitro as well as in vivo latency, and attempts to integrate this knowledge into the framework of the viral life cycle in vivo .

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Primary Immune Responses by Cord Blood CD4+ T Cells and NK Cells Inhibit Epstein-Barr Virus B-Cell Transformation In Vitro

Journal of Virology ◽

10.1128/jvi.76.16.8504.2002 ◽

2002 ◽

Vol 76 (16) ◽

pp. 8504-8504

Author(s):

A. Douglas Wilson ◽

Andrew J. Morgan

Keyword(s):

T Cells ◽

Immune Responses ◽

Cord Blood ◽

B Cell ◽

Nk Cells ◽

Epstein Barr Virus ◽

Cell Transformation ◽

Barr Virus ◽

Epstein Barr

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Epstein-Barr virus nuclear protein 2A forms oligomers in vitro and in vivo through a region required for B-cell transformation.

Journal of Virology ◽

10.1128/jvi.68.7.4287-4294.1994 ◽

1994 ◽

Vol 68 (7) ◽

pp. 4287-4294 ◽

Cited By ~ 13

Author(s):

S Tsui ◽

W H Schubach

Keyword(s):

B Cell ◽

Epstein Barr Virus ◽

Nuclear Protein ◽

Cell Transformation ◽

Barr Virus ◽

Epstein Barr

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Epstein-Barr virus (EBV)-associated lymphoproliferative disease in the SCID mouse model: implications for the pathogenesis of EBV-positive lymphomas in man.

Journal of Experimental Medicine ◽

10.1084/jem.173.1.147 ◽

1991 ◽

Vol 173 (1) ◽

pp. 147-158 ◽

Cited By ~ 191

Author(s):

M Rowe ◽

L S Young ◽

J Crocker ◽

H Stokes ◽

S Henderson ◽

...

Keyword(s):

B Cells ◽

B Cell ◽

Epstein Barr Virus ◽

Large Cell ◽

Burkitt’S Lymphoma ◽

Scid Mice ◽

Burkitt's Lymphoma ◽

Barr Virus ◽

Epstein Barr

When human peripheral blood lymphocytes (PBLs) from Epstein-Barr virus (EBV)-seropositive donors are injected intraperitoneally into SCID mice, EBV+ B cell tumors develop within weeks. A preliminary report (Mosier, D. E., R. J. Gulizia, S. M. Baird, D. D. Richman, D. B. Wilson, R. I. Fox, and T. J. Kipps, 1989. Blood. 74(Suppl. 1):52a) has suggested that such tumors resemble the EBV-positive malignancy, Burkitt's lymphoma. The present work shows that generally the human (hu) PBL-SCID tumors are distinct from Burkitt's lymphoma and instead resemble lymphoblastoid cell lines (LCLs) generated by EBV-infection of normal B cells in vitro in terms of: (a) their cell surface phenotype, with expression of B cell activation antigens and adhesion molecules, (b) normal karyotype, and (c) viral phenotype, with expression of all the transformation-associated EBV latent proteins and, in a minority of cells, productive cycle antigens. Indeed, in vitro-transformed LCLs also grow when inoculated into SCID mice, the frequency of tumor outgrowth correlating with the in vitro growth phenotype of the LCL which is itself determined by the identity of the transforming virus (i.e., type 1 or type 2 EBV). Histologically the PBL-derived hu-SCID tumors resemble the EBV+ large cell lymphomas that develop in immuno-suppressed patients and, like the human tumors, often present at multiple sites as individual monoclonal or oligoclonal foci. The remarkable efficiency of tumor development in the hu-SCID model suggests that lymphomagenesis involves direct outgrowth of EBV-transformed B cells without requirement for secondary genetic changes, and that selection on the basis of cell growth rate alone is sufficient to explain the monoclonal/oligoclonal nature of tumor foci. EBV+ large cell lymphoma of the immunosuppressed may arise in a similar way.

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Epstein-Barr Virus LMP2A Enhances B-Cell Responses In Vivo and In Vitro

Journal of Virology ◽

10.1128/jvi.00433-06 ◽

2006 ◽

Vol 80 (14) ◽

pp. 6764-6770 ◽

Cited By ~ 27

Author(s):

Michelle Swanson-Mungerson ◽

Rebecca Bultema ◽

Richard Longnecker

Keyword(s):

B Cells ◽

B Cell ◽

Epstein Barr Virus ◽

Plasma Cells ◽

Barr Virus ◽

Epstein Barr ◽

Hen Egg ◽

Hen Egg Lysozyme

ABSTRACT Epstein-Barr virus (EBV) establishes latent infections in a significant percentage of the population. Latent membrane protein 2A (LMP2A) is an EBV protein expressed during latency that inhibits B-cell receptor signaling in lymphoblastoid cell lines. In the present study, we have utilized a transgenic mouse system in which LMP2A is expressed in B cells that are specific for hen egg lysozyme (E/HEL-Tg). To determine if LMP2A allows B cells to respond to antigen, E/HEL-Tg mice were immunized with hen egg lysozyme. E/HEL-Tg mice produced antibody in response to antigen, indicating that LMP2A allows B cells to respond to antigen. In addition, E/HEL-Tg mice produced more antibody and an increased percentage of plasma cells after immunization compared to HEL-Tg littermates, suggesting that LMP2A increased the antibody response in vivo. Finally, in vitro studies determined that LMP2A acts directly on the B cell to increase antibody production by augmenting the expansion and survival of the activated B cells, as well as increasing the percentage of plasma cells generated. Taken together, these data suggest that LMP2A enhances, not diminishes, B-cell-specific antibody responses in vivo and in vitro in the E/HEL-Tg system.

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