Time-resolved transcriptomes reveal diverse B cell fate trajectories in the early response to Epstein-Barr virus infection

SoRelle, Elliott D.; Dai, Joanne; Reinoso-Vizcaíno, Nicolás M.; Barry, Ashley P.; Chan, Cliburn; Luftig, Micah A.

doi:10.1016/j.celrep.2022.111286

Cited by 17 publications

(38 citation statements)

References 177 publications

(176 reference statements)

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“…These proliferating cells were consistent with the GC Dark Zone (DZ) state ( 42 , 123 ). In addition to cycling cells, a cluster of a pre-GC activated B cell precursor to early memory B cells (AP-eMBCs) was also identified based on expression of CCR6 , CD22 , and other genes described in prior work ( 3 , 15 , 124 ) ( Figure 3C , S2B ). The four main phenotypes (MBC, AP-eMBC, DZ, and LZ) accounted for 86.4% of cells across LCL datasets.…”

Section: Resultsmentioning

confidence: 88%

“…We recently found that EBV infection induced non-canonical gene expression changes within T-bet + atypical memory B cells (atMBCs) ( 15 ), which have been identified as expanded pathogenic cell subsets in diseases including multiple sclerosis (MS) ( 10 ), systemic lupus erythematosus (SLE) ( 12 , 145 ), and rheumatoid arthritis (RA) ( 146 ). This atMBC response to infection included upregulation of pro-inflammatory signature and aberrant expression of neuronal lineage genes.…”

Section: Resultsmentioning

confidence: 99%

“…This interest was rooted in several distinct lines of evidence. First, we recently reported that de novo EBV infection within atMBCs ( TBX21 /T-bet + , ITGAX /CD11c + , FCRL4 + , FCRL5 + ) induced elevated expression of inflammatory mediators and – unexpectedly – neuronal lineage genes involved in nervous system development and axon guidance ( 15 ). Second, there is strong etiological support for the importance of EBV seropositivity during late adolescence or early adulthood – but not childhood – in the development of MS ( 94 – 96 ).…”

Section: Discussionmentioning

confidence: 99%

“…The widespread utilization of single-cell RNA sequencing (scRNA-seq) has generated new high-dimensional perspectives on the diversity of the human B cell repertoire. Recent scRNA-seq studies have revealed that extensive heterogeneity and dynamic complexity are core aspects of B cell development and clonal selection ( 1 – 4 ), tissue-specific distributions ( 5 ), dysregulation in lymphoma ( 6 – 9 ), pathogenicity in autoimmune disorders ( 10 – 12 ), and responses to antigen ( 13 , 14 ) and infection ( 15 , 16 ). Separately, advances in classic single-cell methods including flow cytometry and cell sorting ( 17 ) provide flexible workflows to validate and isolate cell subsets identified from scRNA-seq experiments and support subsequent investigations.…”

Section: Introductionmentioning

confidence: 99%

“…Our lab has long been interested in dissecting the distinct B cell fates that develop following EBV infection ( 32 , 43 , 107 – 110 ). Recently, we have utilized single-cell sequencing to reveal previously unknown facets of the early stages of infection ( 15 ) and an unexpected degree of heterogeneity in virus-immortalized lymphoblastoid cell lines (LCLs) ( 111 ), which provide useful in vitro models of EBV + lymphomas. This report builds upon our previous single-cell study of LCL heterogeneity to dissect the dynamics of virus-driven B cell responses.…”

Section: Introductionmentioning

confidence: 99%

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Epstein-Barr virus perpetuates B cell germinal center dynamics and generation of autoimmune-associated phenotypes in vitro

et al. 2022

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Human B cells encompass functionally diverse lineages and phenotypic states that contribute to protective as well as pathogenic responses. Epstein-Barr virus (EBV) provides a unique lens for studying heterogeneous B cell responses, given its adaptation to manipulate intrinsic cell programming. EBV promotes the activation, proliferation, and eventual outgrowth of host B cells as immortalized lymphoblastoid cell lines (LCLs) in vitro, which provide a foundational model of viral latency and lymphomagenesis. Although cellular responses and outcomes of infection can vary significantly within populations, investigations that capture genome-wide perspectives of this variation at single-cell resolution are in nascent stages. We have recently used single-cell approaches to identify EBV-mediated B cell heterogeneity in de novo infection and within LCLs, underscoring the dynamic and complex qualities of latent infection rather than a singular, static infection state. Here, we expand upon these findings with functional characterizations of EBV-induced dynamic phenotypes that mimic B cell immune responses. We found that distinct subpopulations isolated from LCLs could completely reconstitute the full phenotypic spectrum of their parental lines. In conjunction with conserved patterns of cell state diversity identified within scRNA-seq data, these data support a model in which EBV continuously drives recurrent B cell entry, progression through, and egress from the Germinal Center (GC) reaction. This “perpetual GC” also generates tangent cell fate trajectories including terminal plasmablast differentiation, which constitutes a replicative cul-de-sac for EBV from which lytic reactivation provides escape. Furthermore, we found that both established EBV latency and de novo infection support the development of cells with features of atypical memory B cells, which have been broadly associated with autoimmune disorders. Treatment of LCLs with TLR7 agonist or IL-21 was sufficient to generate an increased frequency of IgD-/CD27-/CD23-/CD38+/CD138+ plasmablasts. Separately, de novo EBV infection led to the development of CXCR3+/CD11c+/FCRL4+ B cells within days, providing evidence for possible T cell-independent origins of a recently described EBV-associated neuroinvasive CXCR3+ B cell subset in patients with multiple sclerosis. Collectively, this work reveals unexpected virus-driven complexity across infected cell populations and highlights potential roles of EBV in mediating or priming foundational aspects of virus-associated immune cell dysfunction in disease.

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Section: Resultsmentioning

confidence: 88%

Section: Resultsmentioning

confidence: 99%

Section: Discussionmentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

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Epstein-Barr virus perpetuates B cell germinal center dynamics and generation of autoimmune-associated phenotypes in vitro

et al. 2022

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Epstein–Barr virus infection, B‐cell dysfunction and other risk factors converge in gut‐associated lymphoid tissue to drive the immunopathogenesis of multiple sclerosis: a hypothesis

et al. 2022

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Multiple sclerosis is associated with Epstein–Barr virus (EBV) infection, B‐cell dysfunction, gut dysbiosis, and environmental and genetic risk factors, including female sex. A disease model incorporating all these factors remains elusive. Here, we hypothesise that EBV‐infected memory B cells (MBCs) migrate to gut‐associated lymphoid tissue (GALT) through EBV‐induced expression of LPAM‐1, where they are subsequently activated by gut microbes and/or their products resulting in EBV reactivation and compartmentalised anti‐EBV immune responses. These responses involve marginal zone (MZ) B cells that activate CD4 + T‐cell responses, via HLA‐DRB1, which promote downstream B‐cell differentiation towards CD11c + /T‐bet + MBCs, as well as conventional MBCs. Intrinsic expression of low‐affinity B‐cell receptors (BCRs) by MZ B cells and CD11c + /T‐bet + MBCs promotes polyreactive BCR/antibody responses against EBV proteins (e.g. EBNA‐1) that cross‐react with central nervous system (CNS) autoantigens (e.g. GlialCAM). EBV protein/autoantigen‐specific CD11c + /T‐bet + MBCs migrate to the meningeal immune system and CNS, facilitated by their expression of CXCR3, and induce cytotoxic CD8 + T‐cell responses against CNS autoantigens amplified by BAFF, released from EBV‐infected MBCs. An increased abundance of circulating IgA + MBCs, observed in MS patients, might also reflect GALT‐derived immune responses, including disease‐enhancing IgA antibody responses against EBV and gut microbiota‐specific regulatory IgA + plasma cells. Female sex increases MZ B‐cell and CD11c + /T‐bet + MBC activity while environmental risk factors affect gut dysbiosis. Thus, EBV infection, B‐cell dysfunction and other risk factors converge in GALT to generate aberrant B‐cell responses that drive pathogenic T‐cell responses in the CNS.

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