EBNA1-specific CD4+ T cells in healthy carriers of Epstein-Barr virus are primarily Th1 in function

Abstract Abstract 1286 Introduction: T cell expression of PD-1, a marker of functional exhaustion manifested by inability to produce cytokines upon stimulation, is upregulated in patients with acute GHVD. This is thought to explain at least in part why patients with acute GVHD have frequent infections (Gallez-Hawkins et al, BBMT15:872, 2009). Here we wished to evaluate whether this is true also for chronic GVHD. Patients and Methods: We studied 17 allogeneic HCT recipients for AML who have not developed acute or chronic GVHD by day 84. Their blood was drawn on day 84, 180 and 365. Between day 84 and 365, 7 patients did and 10 patients did not develop chronic GVHD needing systemic immunosuppressive therapy (de novo, ie, without preceding acute GVHD). Onset of the chronic GVHD was on median day 103 (range, 90–147). We studied total CD4 and CD8 T cells as well as Epstein-Barr virus (EBV)-specific CD4 and CD8 T cells, as patients with chronic GVHD are at risk of EBV disease (Landgren et al, Blood,14:4992, 2009). Blood mononuclear cells were stimulated with Epstein-Barr virus (EBV) lysate, EBNA3A+B+C overlapping peptides, no or irrelevant stimulus as negative control, or Staphylococcal enterotoxin B as positive control. After overnight incubation, expression of IFNγ, TNFα, IL2 and PD-1 on CD3+CD4+CD8- or CD3+CD4-CD8+ cells was determined by flow cytometry. Cells expressing IFNγ, TNFα, IL2 or their combinations were enumerated. PD-1 expression was quantified using beads coated with anti-mouse antibody (Quantum Simply Cellular, Bangs Laboratories) and expressed as antibody binding capacity units (ABC) (dynamic range, approximately 300 to 500,000 ABC units per cell). Results: PD-1 expression on total, EBV lysate-specific or EBNA3-specific CD4 or CD8 T cells was not significantly higher among patients who did vs did not develop chronic GVHD. On the contrary, there was a trend toward lower PD-1 expression on EBV lysate-specific CD4 and CD8 T cells and EBNA3-specific CD4 T cells in patients who developed chronic GVHD. This was significant (p<.05, Mann-Whitney test) for EBV lysate-specific CD4 T cells on day 84, EBV lysate-specific CD8 T cells on day 180, EBV lysate-specific CD4 and CD8 T cells on day 365, EBNA3-specific CD4 T cells on day 84 and EBNA3-specific CD4 T cells on day 365. Consistent with that, absolute counts of total, EBV lysate-specific or EBNA3-specific T cells were not significantly lower in patients who did vs did not develop chronic GVHD. On the contrary, there was a trend toward higher EBV lysate-specific and EBNA3-specific CD4 or CD8 T cell counts in patients who developed chronic GVHD. This was significant on day 84 for total EBV lysate-specific CD4 and CD8 cells, EBV lysate-specific CD4+IFNγ+ cells and CD8+IFNγ+ cells, and total EBNA3-specific CD4 cells, EBNA3-specific CD4+IFNγ+ cells, CD4+IL2+ cells, CD4+IFNγ+TNFα+IL2+ cells and CD8+IFNγ+ cells. Conclusion: De novo chronic GvHD and its treatment do not adversely affect the counts of functional EBV specific T cells. Disclosures: No relevant conflicts of interest to declare.

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In vitro cytokine production and growth inhibition of lymphoblastoid cell lines by CD4+ T cells from Epstein-Barr virus (EBV) seropositive donors

Clinical & Experimental Immunology ◽

10.1046/j.1365-2249.2001.01641.x ◽

2001 ◽

Vol 126 (1) ◽

pp. 101-110 ◽

Cited By ~ 13

Author(s):

A. D. Wilson ◽

J. C. Hopkins ◽

A. J. Morgan

Keyword(s):

T Cells ◽

Growth Inhibition ◽

Cell Lines ◽

Cd4 T Cells ◽

Epstein Barr Virus ◽

Cytokine Production ◽

Lymphoblastoid Cell Lines ◽

Barr Virus ◽

Epstein Barr

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STAT6 signaling pathway activated by the cytokines IL-4 and IL-13 induces expression of the Epstein-Barr virus–encoded protein LMP-1 in absence of EBNA-2: implications for the type II EBV latent gene expression in Hodgkin lymphoma

Blood ◽

10.1182/blood-2010-01-265272 ◽

2011 ◽

Vol 117 (1) ◽

pp. 165-174 ◽

Cited By ~ 42

Author(s):

Loránd L. Kis ◽

Natalija Gerasimčik ◽

Daniel Salamon ◽

Emma K. Persson ◽

Noémi Nagy ◽

...

Keyword(s):

T Cells ◽

Burkitt Lymphoma ◽

Cd4 T Cells ◽

Epstein Barr Virus ◽

Interleukin 4 ◽

Nuclear Antigen ◽

Type Ii ◽

Signal Transducer ◽

Barr Virus ◽

Epstein Barr

AbstractIn line with the B-lymphotropic nature of Epstein-Barr virus (EBV), the virus is present in several types of B-cell lymphomas. EBV expresses a different set of latent genes in the associated tumors, such as EBV nuclear antigen 1 (EBNA-1) and latent membrane proteins (LMPs; type II latency) in classical Hodgkin lymphomas (HLs). We previously reported that exposure of in vitro EBV-converted, HL-derived cell line KMH2-EBV to CD40-ligand and interleukin-4 (IL-4) induced the expression of LMP-1. Here, we show that exposure to IL-4 or IL-13 alone induced LMP-1 in the absence of EBNA-2. Induction of LMP-1 by IL-4 and IL-13 was mediated by the signal transducer signal transducer and activator of transcription 6 (STAT6) and a newly defined high-affinity STAT6-binding site in the LMP-1 promoter. IL-4 induced LMP-1 also in Burkitt lymphoma–derived lines and in tonsillar B cells infected with the EBNA-2–deficient EBV strain P3HR-1. Furthermore, coculture of EBV-carrying Burkitt lymphoma cells with activated CD4+ T cells resulted in the induction of LMP-1 in the absence of EBNA-2. Because Hodgkin/Reed-Sternberg cells are known to secrete IL-13, to have constitutively activated STAT6, and to be closely surrounded by CD4+ T cells, these mechanisms may be involved in the expression of LMP-1 in EBV-positive chronic HLs.

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INFLUENCE OF EPSTEIN BARR VIRUS (EBV) EXPRESSION ON CD4+ T CELLS AT THE MICROENVIRONMENT IN PEDIATRIC HODGKIN LYMPHOMA

Hematological Oncology ◽

10.1002/hon.8_2881 ◽

2021 ◽

Vol 39 (S2) ◽

Author(s):

O Jimenez ◽

M. V Preciado ◽

E. De Matteo ◽

P Chabay

Keyword(s):

T Cells ◽

Hodgkin Lymphoma ◽

Cd4 T Cells ◽

Epstein Barr Virus ◽

Barr Virus ◽

Pediatric Hodgkin Lymphoma ◽

Epstein Barr

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Epstein-Barr virus nuclear antigen 1-specific CD4+T cells directly kill Epstein-Barr virus-carrying natural killer and T cells

Cancer Science ◽

10.1111/j.1349-7006.2008.00852.x ◽

2008 ◽

Vol 99 (8) ◽

pp. 1633-1642 ◽

Cited By ~ 24

Author(s):

Ayako Demachi-Okamura ◽

Yoshinori Ito ◽

Yoshiki Akatsuka ◽

Kunio Tsujimura ◽

Yasuo Morishima ◽

...

Keyword(s):

T Cells ◽

Natural Killer ◽

Cd4 T Cells ◽

Epstein Barr Virus ◽

Nuclear Antigen ◽

Barr Virus ◽

Epstein Barr

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Distinct memory CD4+ T-cell subsets mediate immune recognition of Epstein Barr virus nuclear antigen 1 in healthy virus carriers

Blood ◽

10.1182/blood-2006-05-023663 ◽

2006 ◽

Vol 109 (3) ◽

pp. 1138-1146 ◽

Cited By ~ 43

Author(s):

Kevin N. Heller ◽

Jenica Upshaw ◽

Beza Seyoum ◽

Henry Zebroski ◽

Christian Münz

Keyword(s):

T Cells ◽

Cd4 T Cells ◽

Epstein Barr Virus ◽

T Cell Subsets ◽

Nuclear Antigen ◽

Effector Memory ◽

Immune Recognition ◽

T Helper 1 ◽

Barr Virus ◽

Epstein Barr

AbstractCD4+ T cells, specific for transforming latent infection with the Epstein Barr virus (EBV), consistently recognize the nuclear antigen 1 of EBV (EBNA1). EBNA1-specific effector CD4+ T cells are primarily T-helper 1 (TH1) polarized. Here we show that most healthy EBV carriers have such IFN-secreting EBNA1-specific CD4+ T cells at a frequency of 0.03% of circulating CD4+ T cells. In addition, healthy carriers have a large pool of CD4+ T cells that proliferated in response to EBNA1 and consisted of distinct memory-cell subsets. Despite continuous antigen presence due to persistent EBV infection, half of the proliferating EBNA1-specific CD4+ T cells belonged to the central-memory compartment (TCM). The remaining EBNA1-specific CD4+ T cells displayed an effector-memory phenotype (TEM), of which a minority rapidly secreted IFN upon stimulation with EBNA1. Based on chemokine receptor analysis, all EBNA1-specific TCM CD4+ T cells were TH1 committed. Our results suggest that protective immune control of chronic infections, like EBV, includes a substantial reservoir of TCM CD4+ TH1 precursors, which continuously fuels TH1-polarized effector cells.

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Human Cd4+ T Lymphocytes Consistently Respond to the Latent Epstein-Barr Virus Nuclear Antigen Ebna1

Journal of Experimental Medicine ◽

10.1084/jem.191.10.1649 ◽

2000 ◽

Vol 191 (10) ◽

pp. 1649-1660 ◽

Cited By ~ 238

Author(s):

Christian Münz ◽

Kara L. Bickham ◽

Marion Subklewe ◽

Ming L. Tsang ◽

Ann Chahroudi ◽

...

Keyword(s):

T Cells ◽

T Cell ◽

B Cells ◽

T Lymphocytes ◽

Cd4 T Cells ◽

Epstein Barr Virus ◽

Cd4 T Cell ◽

Nuclear Antigen ◽

Barr Virus ◽

Epstein Barr

The Epstein-Barr virus (EBV)-encoded nuclear antigen EBNA1 is critical for the persistence of the viral episome in replicating EBV-transformed human B cells. Therefore, all EBV-induced tumors express this foreign antigen. However, EBNA1 is invisible to CD8+ cytotoxic T lymphocytes because its Gly/Ala repeat domain prevents proteasome-dependent processing for presentation on major histocompatibility complex (MHC) class I. We now describe that CD4+ T cells from healthy adults are primed to EBNA1. In fact, among latent EBV antigens that stimulate CD4+ T cells, EBNA1 is preferentially recognized. We present evidence that the CD4+ response may provide a protective role, including interferon γ secretion and direct cytolysis after encounter of transformed B lymphocyte cell lines (B-LCLs). Dendritic cells (DCs) process EBNA1 from purified protein and from MHC class II–mismatched, EBNA1-expressing cells including B-LCLs. In contrast, B-LCLs and Burkitt's lymphoma lines likely present EBNA1 after endogenous processing, as their capacity to cross-present from exogenous sources is weak or undetectable. By limiting dilution, there is a tight correlation between the capacity of CD4+ T cell lines to recognize autologous B-LCL–expressing EBNA1 and DCs that have captured EBNA1. Therefore, CD4+ T cells can respond to the EBNA1 protein that is crucial for EBV persistence. We suggest that this immune response is initiated in vivo by DCs that present EBV-infected B cells, and that EBNA1-specific CD4+ T cell immunity be enhanced to prevent and treat EBV-associated malignancies.

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CD4 + T cells are found within endemic Burkitt lymphoma and modulate Burkitt lymphoma precursor cell viability and expression of pathogenically relevant Epstein–Barr virus genes

Cancer Immunology Immunotherapy ◽

10.1007/s00262-021-03057-5 ◽

2021 ◽

Author(s):

Semjon Sidorov ◽

Lara Fux ◽

Katja Steiner ◽

Samyo Bounlom ◽

Sabrina Traxel ◽

...

Keyword(s):

T Cells ◽

T Cell ◽

Burkitt Lymphoma ◽

Cd4 T Cells ◽

Epstein Barr Virus ◽

Cell Cancer ◽

Cd4 T Cell ◽

Barr Virus ◽

Epstein Barr

AbstractEndemic Burkitt lymphoma (eBL) is an aggressive B cell cancer characterized by an IgH/c-myc translocation and the harboring of Epstein–Barr virus (EBV). Evidence accumulates that CD4 + T cells might contribute to eBL pathogenesis. Here, we investigate the presence of CD4 + T cells in primary eBL tissue and their potential dichotomous impact on an EBV-infected pre-eBL cell model using ex vivo material and in vitro co-cultures. In addition, we establish a novel method to study the effect of IgH/c-myc translocation in primary B cells by employing a CRISPR/Cas9 knock-in approach to introduce and tag de novo translocation. We unprecedently document that CD4 + T cells are present in primary eBL tumor tissue. Furthermore, we demonstrate that CD4 + T cells on the one hand suppress eBL development by killing pre-eBL cells lacking IgH/c-myc translocation in vitro and on the other hand indirectly promote eBL development by inducing crucial EBV Latency III to Latency I switching in pre-eBL cells. Finally, we show that while the mere presence of an IgH/c-myc translocation does not suffice to escape CD4 + T-cell-mediated killing in vitro, the CD4 + T-cell-mediated suppression of EBV’s Latency III program in vivo may allow cells harboring an IgH/c-myc translocation and additional mutations to evade immune control and proliferate by means of deregulated c-myc activity, resulting in neoplasia. Thus, our study highlights the dichotomous effects of CD4 + T cells and the mechanisms involved in eBL pathogenesis, suggests mechanisms of their impact on eBL progression, and provides a novel in vitro model for further investigation of IgH/c-myc translocation.

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Epstein-Barr Virus–induced Molecule 1 Ligand Chemokine Is Expressed by Dendritic Cells in Lymphoid Tissues and Strongly Attracts Naive T Cells and Activated B Cells

Journal of Experimental Medicine ◽

10.1084/jem.188.1.181 ◽

1998 ◽

Vol 188 (1) ◽

pp. 181-191 ◽

Cited By ~ 339

Author(s):

Vu N. Ngo ◽

H. Lucy Tang ◽

Jason G. Cyster

Keyword(s):

T Cells ◽

Dendritic Cells ◽

B Cells ◽

Cd4 T Cells ◽

Epstein Barr Virus ◽

Cell Receptor ◽

Lymphoid Tissues ◽

Barr Virus ◽

Epstein Barr ◽

Activated B Cells

Movement of T and B lymphocytes through secondary lymphoid tissues is likely to involve multiple cues that help the cells navigate to appropriate compartments. Epstein-Barr virus– induced molecule 1 (EBI-1) ligand chemokine (ELC/MIP3β) is expressed constitutively within lymphoid tissues and may act as such a guidance cue. Here, we have isolated mouse ELC and characterized its expression pattern and chemotactic properties. ELC is expressed constitutively in dendritic cells within the T cell zone of secondary lymphoid tissues. Recombinant ELC was strongly chemotactic for naive (L-selectinhi) CD4 T cells and for CD8 T cells and weakly attractive for resting B cells and memory (L-selectinlo) CD4 T cells. After activation through the B cell receptor, the chemotactic response of B cells was enhanced. Like its human counterpart, murine ELC stimulated cells transfected with EBI-1/CC chemokine receptor 7 (CCR7). Our findings suggest a central role for ELC in promoting encounters between recirculating T cells and dendritic cells and in the migration of activated B cells into the T zone of secondary lymphoid tissues.

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