Cytotoxic T lymphocyte immunodominance to Epstein-Barr virus infection

EBV lytic replication involves the sequential expression of a large array of antigens that potentially provides a complex antigenic challenge. Despite this number, the primary CD8+ T cell response in infectious mononucleosis (IM) patients appears focused against epitopes found in immediate-early and some early expressed antigens with responses to late antigens typically subdominant. However previous approaches have only focused on a limited number of lytic antigens. To resolve these issues, this study examines the CD8+ T cell repertoire to all EBY lytic antigens in different phases of infection. Polyelonal CD8+ T cell lines were mitogenically expanded from IM patients or expanded in an antigenically unbiased way from post-1M patients and healthy carriers using autologous dendritic cells loaded with lysates of lytically infected cells. Target cells expressing individual lytic antigens along with donor HLA class I molecules were used to challenge polyclonal lines with responses measured by the release of IFNγ. These studies show that the pattern of target antigen choice varies with the phase of infection and is consistent with the idea that CD8+ T cell responses in primary infection are driven by lytically infected B cells. However over time the repertoire of responses may be influenced through antigen cross-presentation

Diseases of Pyrethrum in Tasmania: Challenges and Prospects for Management

Pyrethrum (Tanacetum cinerariifolium (Trevir.) Sch. Bip.) is a perennial plant and member of the Asteraceae that is endemic to the Dalmatian region of the former Yugoslavia (36). Pyrethrum is cultivated commercially solely for the production of six closely related esters called pyrethrins. The plant is tufted, slender, and herbaceous, growing to a height of approximately one meter (18). Leaves are alternate and pinnately lobed/narrowly lanceolate to oblong lanceolate. The daisy-like flowers are produced at the termini of stems and consist of a cluster of 40 to 100 bisexual, yellow disk florets encircled by a ring of 18 to 22 pistillate white ray florets atop a moderately convex to subglobose receptacle (Fig. 1; 100). Disk and ray florets both possess 3 to 10 ribbed achenes located between the floret and receptacle. Involucres generally range between 12 and 18 mm in diameter (17,18). Approximately 94% o

Localization of Double-Strand Break Repair Proteins to Viral Replication Compartments following Lytic Reactivation of Kaposi's Sarcoma-Associated Herpesvirus

ABSTRACT Double-strand breaks (DSBs) in DNA are recognized by the Ku70/80 heterodimer and the MRE11-RAD50-NBS1 (MRN) complex and result in activation of the DNA-PK and ATM kinases, which play key roles in regulating the cellular DNA damage response (DDR). DNA tumor viruses such as Kaposi's sarcoma-associated herpesvirus (KSHV) are known to interact extensively with the DDR during the course of their replicative cycles. Here we show that during lytic amplification of KSHV DNA, the Ku70/80 heterodimer and the MRN complex consistently colocalize with viral genomes in replication compartments (RCs), whereas other DSB repair proteins form foci outside RCs. Depletion of MRE11 and abrogation of its exonuclease activity negatively impact viral replication, while in contrast, knockdown of Ku80 and inhibition of the DNA-PK enzyme, which are involved in nonhomologous end joining (NHEJ) repair, enhance amplification of viral DNA. Although the recruitment of DSB-sensing proteins to KSHV RCs is a consistent occurrence across multiple cell types, activation of the ATM-CHK2 pathway during viral replication is a cell line-specific event, indicating that recognition of viral DNA by the DDR does not necessarily result in activation of downstream signaling pathways. We have also observed that newly replicated viral DNA is not associated with cellular histones. Since the presence and modification of these DNA-packaging proteins provide a scaffold for docking of multiple DNA repair factors, the absence of histone deposition may allow the virus to evade localization of DSB repair proteins that would otherwise have a detrimental effect on viral replication. IMPORTANCE Tumor viruses are known to interact with machinery responsible for detection and repair of double-strand breaks (DSBs) in DNA, although detail concerning how Kaposi's sarcoma-associated herpesvirus (KSHV) modulates these cellular pathways during its lytic replication phase was previously lacking. By undertaking a comprehensive assessment of the localization of DSB repair proteins during KSHV replication, we have determined that a DNA damage response (DDR) is directed to viral genomes but is distinct from the response to cellular DNA damage. We also demonstrate that although recruitment of the MRE11-RAD50-NBS1 (MRN) DSB-sensing complex to viral genomes and activation of the ATM kinase can promote KSHV replication, proteins involved in nonhomologous end joining (NHEJ) repair restrict amplification of viral DNA. Overall, this study extends our understanding of the virus-host interactions that occur during lytic replication of KSHV and provides a deeper insight into how the DDR is manipulated during viral infection. </jats:p

Proteome-wide analysis of CD8+ T cell responses to EBV reveals differences between primary and persistent infection

Human herpesviruses are antigenically rich agents that induce strong CD8+T cell responses in primary infection yet persist for life, continually challenging T cell memory through recurrent lytic replication and potentially influencing the spectrum of antigen-specific responses. Here we describe the first lytic proteome-wide analysis of CD8+ T cell responses to a gamma1-herpesvirus, Epstein-Barr virus (EBV), and the first such proteome-wide analysis of primary versus memory CD8+ T cell responses to any human herpesvirus. Primary effector preparations were generated directly from activated CD8+ T cells in the blood of infectious mononucleosis (IM) patients by in vitro mitogenic expansion. For memory preparations, EBV-specific cells in the blood of long-term virus carriers were first re-stimulated in vitro by autologous dendritic cells loaded with a lysate of lytically-infected cells, then expanded as for IM cells. Preparations from 7 donors of each type were screened against each of 70 EBV lytic cycle proteins in combination with the donor's individual HLA class I alleles. Multiple reactivities against immediate early (IE), early (E) and late (L) lytic cycle proteins, including many hitherto unrecognised targets, were detected in both contexts. Interestingly however, the two donor cohorts showed a different balance between IE, E and L reactivities. Primary responses targeted IE and a small group of E proteins preferentially, seemingly in line with their better presentation on the infected cell surface before later-expressed viral evasins take full hold. By contrast, target choice equilibrates in virus carriage with responses to key IE and E antigens still present but with responses to a select subset of L proteins now often prominent. We infer that, for EBV at least, long-term virus carriage with its low level virus replication and lytic antigen release is associated with a re-shaping of the virus-specific response

Control of human cytomegalovirus replication by liver resident natural killer cells

Natural killer cells are considered to be important for control of human cytomegalovirus– a major pathogen in immune suppressed transplant patients. Viral infection promotes the development of an adaptive phenotype in circulating natural killer cells that changes their anti-viral function. In contrast, less is understood how natural killer cells that reside in tissue respond to viral infection. Here we show natural killer cells resident in the liver have an altered phenotype in cytomegalovirus infected individuals and display increased anti-viral activity against multiple viruses in vitro and identify and characterise a subset of natural killer cells responsible for control. Crucially, livers containing natural killer cells with better capacity to control cytomegalovirus replication in vitro are less likely to experience viraemia post-transplant. Taken together, these data suggest that virally induced expansion of tissue resident natural killer cells in the donor organ can reduce the chance of viraemia post-transplant

The missing link in EBV immune evasion:the BDLF3 gene induces ubiquitination and downregulation of MHC class I and MHC class II

The ability of Epstein-Barr virus (EBV) to spread and persist in human populations relies on a balance between host immune responses and EBV immune evasion. CD8(+) cells specific for EBV late lytic cycle antigens show poor recognition of target cells compared to immediate early and early antigen-specific CD8(+) cells. This phenomenon is due in part to the early EBV protein BILF1, whose immunosuppressive activity increases with lytic cycle progression. However, published data suggest the existence of a hitherto unidentified immune evasion protein further enhancing protection against late EBV antigen-specific CD8(+) cells. We have now identified the late lytic BDLF3 gene as the missing link accounting for efficient evasion during the late lytic cycle. Interestingly, BDLF3 also contributes to evasion of CD4(+) cell responses to EBV. We report that BDLF3 downregulates expression of surface major histocompatibility complex (MHC) class I and class II molecules in the absence of any effect upon other surface molecules screened, including CD54 (ICAM-1) and CD71 (transferrin receptor). BDLF3 both enhanced internalization of surface MHC molecules and reduced the rate of their appearance at the cell surface. The reduced expression of surface MHC molecules correlated with functional protection against CD8(+) and CD4(+) T cell recognition. The molecular mechanism was identified as BDLF3-induced ubiquitination of MHC molecules and their subsequent downregulation in a proteasome-dependent manner. IMPORTANCE Immune evasion is a necessary feature of viruses that establish lifelong persistent infections in the face of strong immune responses. EBV is an important human pathogen whose immune evasion mechanisms are only partly understood. Of the EBV immune evasion mechanisms identified to date, none could explain why CD8(+) T cell responses to late lytic cycle genes are so infrequent and, when present, recognize lytically infected target cells so poorly relative to CD8(+) T cells specific for early lytic cycle antigens. The present work identifies an additional immune evasion protein, BDLF3, that is expressed late in the lytic cycle and impairs CD8(+) T cell recognition by targeting cell surface MHC class I molecules for ubiquitination and proteasome-dependent downregulation. Interestingly, BDLF3 also targets MHC class II molecules to impair CD4(+) T cell recognition. BDLF3 is therefore a rare example of a viral protein that impairs both the MHC class I and class II antigen-presenting pathways

An earth system governance research agenda for carbon removal

Carbon dioxide removal (CDR) – the creation, enhancement, and upscaling of carbon sinks – has become a pillar of national and corporate commitments towards Net Zero emissions, as well as pathways towards realizing the Paris Agreement's ambitious temperature targets. In this perspective, we explore CDR as an emerging issue of Earth System Governance (ESG). We draw on the results of a workshop at the 2022 Earth System Governance conference that mapped a range of actors, activities, and issues relevant to carbon removal, and refined them into research questions spanning four intersecting areas: modeling and systems assessment, societal appraisal, policy, and innovation and industry. We filter these questions through the five lenses of the ESG framework and highlight several key ‘cross-cutting’ issues that could form the basis of an integrated ESG research agenda on CDR

An earth system governance research agenda for carbon removal

Carbon dioxide removal (CDR) – the creation, enhancement, and upscaling of carbon sinks – has become a pillar of national and corporate commitments towards Net Zero emissions, as well as pathways towards realizing the Paris Agreement's ambitious temperature targets. In this perspective, we explore CDR as an emerging issue of Earth System Governance (ESG). We draw on the results of a workshop at the 2022 Earth System Governance conference that mapped a range of actors, activities, and issues relevant to carbon removal, and refined them into research questions spanning four intersecting areas: modeling and systems assessment, societal appraisal, policy, and innovation and industry. We filter these questions through the five lenses of the ESG framework and highlight several key ‘cross-cutting’ issues that could form the basis of an integrated ESG research agenda on CDR.</p

An earth system governance research agenda for carbon removal

Carbon dioxide removal (CDR) – the creation, enhancement, and upscaling of carbon sinks – has become a pillar of national and corporate commitments towards Net Zero emissions, as well as pathways towards realizing the Paris Agreement's ambitious temperature targets. In this perspective, we explore CDR as an emerging issue of Earth System Governance (ESG). We draw on the results of a workshop at the 2022 Earth System Governance conference that mapped a range of actors, activities, and issues relevant to carbon removal, and refined them into research questions spanning four intersecting areas: modeling and systems assessment, societal appraisal, policy, and innovation and industry. We filter these questions through the five lenses of the ESG framework and highlight several key ‘cross-cutting’ issues that could form the basis of an integrated ESG research agenda on CDR

Search for dark matter produced in association with bottom or top quarks in √s = 13 TeV pp collisions with the ATLAS detector

A search for weakly interacting massive particle dark matter produced in association with bottom or top quarks is presented. Final states containing third-generation quarks and miss- ing transverse momentum are considered. The analysis uses 36.1 fb−1 of proton–proton collision data recorded by the ATLAS experiment at √s = 13 TeV in 2015 and 2016. No significant excess of events above the estimated backgrounds is observed. The results are in- terpreted in the framework of simplified models of spin-0 dark-matter mediators. For colour- neutral spin-0 mediators produced in association with top quarks and decaying into a pair of dark-matter particles, mediator masses below 50 GeV are excluded assuming a dark-matter candidate mass of 1 GeV and unitary couplings. For scalar and pseudoscalar mediators produced in association with bottom quarks, the search sets limits on the production cross- section of 300 times the predicted rate for mediators with masses between 10 and 50 GeV and assuming a dark-matter mass of 1 GeV and unitary coupling. Constraints on colour- charged scalar simplified models are also presented. Assuming a dark-matter particle mass of 35 GeV, mediator particles with mass below 1.1 TeV are excluded for couplings yielding a dark-matter relic density consistent with measurements