Abstract
Subpopulations of B-lymphocytes traffic to different sites and organs to provide diverse and tissue-specific functions. Here, we provide evidence that epigenetic differences confer a neuroinvasive phenotype. An EBV+ B cell lymphoma cell line (M14) with low frequency trafficking to the CNS was neuroadapted to generate a highly neuroinvasive B-cell population (MUN14). MUN14 B cells efficiently infiltrated the CNS within one week and produced neurological pathologies. We compared the gene expression profiles of viral and cellular genes using RNA-Seq and identified one viral (EBNA1) and several cellular gene candidates, including secreted phosphoprotein 1/osteopontin (SPP1/OPN), neuron navigator 3 (NAV3), CXCR4, and germinal center-associated signaling and motility protein (GCSAM) that were selectively upregulated in MUN14. ATAC-Seq and ChIP-qPCR revealed that these gene expression changes correlated with epigenetic changes at gene regulatory elements. The neuroinvasive phenotype could be attenuated with a neutralizing antibody to OPN, confirming the functional role of this protein in trafficking EBV+ B cells to the CNS. These studies indicate that B-cell trafficking to the CNS can be acquired by epigenetic adaptations and provide a new model to study B-cell neuroinvasion associated CNS lymphoma and autoimmune disease of the CNS, including multiple sclerosis (MS).
Highlights
Epstein-Barr virus (EBV) is a near ubiquitous gamma-herpesvirus that establishes latent infection in long-lived B-lymphocytes and is associated with diverse human disease, including cancer and auto-immune disorders [1,2,3]
We show that EBV+ B-cells can undergo an epigenetic switch conferring a neuroinvasive phenotype with upregulation of secreted phosphoprotein 1 (SPP1)/Osteopontin, which when blocked reduces CNS penetrance
To better understand EBV-encoded and cellular factors that promote CNS penetration of EBV+ B cells, we developed a mouse model to study the infiltration of EBV immortalized B cells in the CNS using a neuroadapted, EBV+ B lymphoma line (MUN14) with an increased neuroinvasive phenotype compared to the parent line (M14)
Summary
Epstein-Barr virus (EBV) is a near ubiquitous gamma-herpesvirus that establishes latent infection in long-lived B-lymphocytes and is associated with diverse human disease, including cancer and auto-immune disorders [1,2,3]. The virus transiently infects epithelial and lymphoid cells in the Waldeyer’s ring of the oropharynx but undergoes a stable transformative infection in B cells expressing CD21, the major cellular receptor for EBV [6,7,8]. The virus induces developmental changes in resting B-cells that mimic germinal center reactions and memory B-cell phenotype (IgD-CD27+ and IgD+CD27+) where the virus establishes life-long infection [9,10]. During this B-cell transformation process, both EBV and cellular genomes undergo varying degrees of epigenetic modification, including silencing of many viral genes necessary to establish stable latent infection [11,12,13]. EBV is found in only ~1 per million circulating B-cells, but virus may reside in B-cells at diverse stages of differentiation in lymphoid organs [16]
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