Distinct Ex Vivo Susceptibility of B-Cell Subsets to Epstein-Barr Virus Infection According to Differentiation Status and Tissue Origin (original) (raw)
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EBV Persistence in Memory B Cells In Vivo
Immunity, 1998
latency in resting cells. * Department of Pathology The presence of latent EBV in resting B cells creates Tufts University School of Medicine a paradox-how does EBV, which drives infected cells Boston, Massachusetts 02111 to proliferate in vitro, manage to persist in a resting cell † Department of Otolaryngology in vivo? There are two possibilities. Either the virus is New England Medical Center Hospital able to establish latency directly in resting B cells in Boston, Massachusetts 02111 vivo, or the newly infected cells become activated and proliferate but are able to exit from the cell cycle. Several arguments favor the latter explanation. First, the viral Summary DNA in the resting B cells is a covalently closed circular episome (Decker et al., 1996) that forms in vitro when Epstein-Barr virus establishes latency in vitro by actian infected cell becomes activated and enters the cell vating human B cells to become proliferating blasts, cycle (Hurley and Thorley-Lawson, 1988). Therefore, the but in vivo it is benign. In the peripheral blood, the resting cells must have been proliferating at some point virus resides latently in resting B cells that we now in time. Second, the transcriptional program of EBV is show are restricted to the sIgD Ϫ memory subset. Howcomplex and appears to be specifically designed to ever, in tonsils the virus shows no such restriction. efficiently activate newly infected B cells and drive them We propose that EBV indiscriminately infects B cells into the cell cycle. It is unclear how EBV could enter a in mucosal lymphoid tissue and that these cells differresting B cell and quiescently establish latency without entiate to become resting memory B cells that then initiating this program, but if it could the activation mechenter the circulation. Activation to the blastoid stage anism would be unnecessary. In fact, the ability to actiof latency is an essential intermediate step in this provate B cells has been retained even though it poses cess. Thus, EBV may persist by exploiting the mechaa threat, predisposition to neoplasia, to the very host nisms that produce and maintain long-term B cell organism that the virus depends on for long-term persismemory. tence and survival. Therefore, EBV must have evolved a mechanism for B cell activation because it is an essential
Immunology, 2003
While Epstein±Barr virus (EBV) is known to establish latency in the memory B-cell compartment, there is controversy as to whether the memory or the naõ Ève B cell is the initial target for infection. Here we have explored the infectability of the B-cell subsets contained in peripheral blood and tonsils, as distinguished by their surface expression of the immunoglobulin isotypes that help to de®ne naõ Ève and memory pools. First we show that both CD21 and major histocompatibility complex (MHC) class II molecules ± respectively, the major receptor and co-receptor for EBVon B cells ± are expressed at similar levels on blood and tonsillar B cells, irrespective of surface immunoglobulin class, indicating that each of the subsets demonstrate an equal potential, at least for infection. Then, following in vitro infection of total tonsillar B cells, we found that the relative frequencies of immunoglobulin (Ig)M-, IgG-and IgA-positive cells containing EBV-encoded Epstein±Barr virus nuclear antigen 5 (EBNA5) protein at 48 hr were similar to those of the starting population. However, IgD expression was uniformly decreased, probably as a consequence of cellular activation. These data indicate that recirculating B cells have both the potential for, and susceptibility to, initial infection by EBV, irrespective of the immunoglobulin isotype expressed.
Epstein-Barr Virus Can Establish Infection in the Absence of a Classical Memory B-Cell Population
Journal of Virology, 2005
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Is EBV Persistence In Vivo a Model for B Cell Homeostasis?
Immunity, 1996
the assays used for the PCR studies were only sensitive enough to measure the relative genome copy number. Bin Yang, Gregory J. Babcock, However, it is known that latently infected cells in vitro and David A. Thorley-Lawson can contain multiple copies of the viral genome that can Deparment of Pathology range from 5-500 and that cells Tufts University School of Medicine replicating the virus contain thousands of genomes Boston, Massachusetts 02111 (Summers and Klein, 1976). Thus, the PCR assays could only compare relative burdens of viral DNA. They could not measure the actual numbers of infected cells in an Summary individual and could not, therefore, be used meaningfully to compare measurements of the number of infected We have measured the absolute numbers of EBVcells either within or between individuals. For example, infected B cells in the peripheral blood of healthy perit is well established that the viral genome burden insistently infected individuals. Single measurements on creases in immunosuppressed individuals (Saito et al., a panel of 15 healthy individuals demonstrate that the 1989; Telenti et al. , 1990). But the PCR-based assays frequency varies over a wide range from 1-50 per 10 6
Early Events Associated with Infection of Epstein-Barr Virus Infection of Primary B-Cells
PLoS ONE, 2009
Epstein Barr virus (EBV) is closely associated with the development of a vast number of human cancers. To develop a system for monitoring early cellular and viral events associated with EBV infection a self-recombining BAC containing 172-kb of the Epstein Barr virus genome BAC-EBV designated as MD1 BAC (Chen et al., 2005, J.Virology) was used to introduce an expression cassette of green fluorescent protein (GFP) by homologous recombination, and the resultant BAC clone, BAC-GFP-EBV was transfected into the HEK 293T epithelial cell line. The resulting recombinant GFP EBV was induced to produce progeny virus by chemical inducer from the stable HEK 293T BAC GFP EBV cell line and the virus was used to immortalize human primary B-cell as monitored by green fluorescence and outgrowth of the primary B cells. The infection, B-cell activation and cell proliferation due to GFP EBV was monitored by the expression of the B-cell surface antigens CD5, CD10, CD19, CD23, CD39, CD40 , CD44 and the intercellular proliferation marker Ki-67 using Flow cytometry. The results show a dramatic increase in Ki-67 which continues to increase by 6-7 days post-infection. Likewise, CD40 signals showed a gradual increase, whereas CD23 signals were increased by 6-12 hours, maximally by 3 days and then decreased. Monitoring the viral gene expression pattern showed an early burst of lytic gene expression. This up-regulation of lytic gene expression prior to latent genes during early infection strongly suggests that EBV infects primary B-cell with an initial burst of lytic gene expression and the resulting progeny virus is competent for infecting new primary B-cells. This process may be critical for establishment of latency prior to cellular transformation. The newly infected primary B-cells can be further analyzed for investigating B cell activation due to EBV infection.
Persistence of Epstein-Barr Virus in Self-Reactive Memory B Cells
Journal of Virology, 2012
Epstein-Barr virus infection has been epidemiologically associated with the development of multiple autoimmune diseases, particularly systemic lupus erythematosus and multiple sclerosis. Currently, there is no known mechanism that can account for these associations. The germinal-center (GC) model of EBV infection and persistence proposes that EBV gains access to the memory B cell compartment via GC reactions by driving infected cells to differentiate using the virus-encoded LMP1 and LMP2a proteins, which act as functional homologues of CD40 and the B cell receptor, respectively. The ability of LMP2a, when expressed in mice, to allow escape of autoreactive B cells suggests that it could perform a similar role in infected GC B cells, permitting the survival of potentially pathogenic autoreactive B cells. To test this hypothesis, we cloned and expressed antibodies from EBV ؉ and EBV ؊ memory B cells present during acute infection and profiled their self-and polyreactivity. We find that EBV does persist within self-and polyreactive B cells but find no evidence that it favors the survival of pathogenic autoreactive B cells. On the contrary, EBV ؉ memory B cells express lower levels of self-reactive and especially polyreactive antibodies than their uninfected counterparts do. Our work suggests that EBV has only a modest effect on the GC process, which allows it to access and persist within a subtly unique niche of the memory compartment characterized by relatively low levels of self-and polyreactivity. We suggest that this might reflect an active process where EBV and its human host have coevolved so as to minimize the virus's potential to contribute to autoimmune disease.
Virology journal, 2011
Epstein-Barr virus (EBV)-driven B cell proliferation is critical to its subsequent persistence in the host and is a key event in the development of EBV-associated B cell diseases. Thus, inquiry into early cellular events that precede EBV-driven proliferation of B cells is essential for understanding the processes that can lead to EBV-associated B cell diseases. Infection with high titers of EBV of mixed, primary B cells in different stages of differentiation occurs during primary EBV infection and in the setting of T cell-immunocompromise that predisposes to development of EBV-lymphoproliferative diseases. Using an ex vivo system that recapitulates these conditions of infection, we correlated expression of selected B cell-surface markers and intracellular cytokines with expression of EBV latency genes and cell proliferation. We identified CD23, CD58, and IL6, as molecules expressed at early times after EBV-infection. EBV differentially infected B cells into two distinct sub-populati...
Proceedings of the National Academy of Sciences, 2005
Epstein–Barr virus (EBV) establishes a lifelong persistent infection within peripheral blood B cells with the surface phenotype of memory cells. To date there is no proof that these cells have the genotype of true germinal-center-derived memory B cells. It is critical to understand the relative contribution of viral mimicry versus antigen signaling to the production of these cells because EBV encodes proteins that can affect the surface phenotype of infected cells and provide both T cell help and B cell receptor signals in the absence of cognate antigen. To address these questions we have developed a technique to identify single EBV-infected cells in the peripheral blood and examine their expressed Ig genes. The genes were all isotype-switched and somatically mutated. Furthermore, the mutations do not cause stop codons and display the pattern expected for antigen-selected memory cells based on their frequency, type, and location within the Ig gene. We conclude that latently infected...