Ectopic lymphoid structures support Epstein-Barr virus persistence and autoreactive plasma cell infection in Rheumatoid Arthritis synovium and Sjogren’s Syndrome salivary glands

PhDThe ubiquitous ɣ-herpesvirus Epstein-Barr virus (EBV) infects B cells and modifies their differentiation programme leading to B cell activation and immortalization. Although different evidences support a link between EBV infection and rheumatoid arthritis (RA) and Sjogren’s syndrome (SS), the exact role of EBV in RA and SS pathogenesis remain elusive. Recently ectopic lymphoid structures (ELS) have been identified as preferential niches for EBV persistence and reactivation in patients with multiple sclerosis and myasthenia gravis. Independent studies demonstrated that around 50% of RA synovia and 30% of SS salivary glands are characterised by the development of functional ELS, capable to promote local differentiation of autoreactive plasma cells. In this PhD project I explored the potential role of EBV in RA and SS pathogenesis by analysing EBV infection in the RA synovium and SS salivary glands and its relationship with ELS, in situ autoreactive plasma cell differentiation, pathogenic autoantibodies production and cytotoxic immune response. In this work I demonstrated that: i) markers of EBV latent and lytic infection are consistently associated with the presence of ELS in the RA synovium and SS salivary glands; ii) latent EBV proteins are preferentially expressed by B cells, while viral reactivation occurs in plasma cells; iii) a large subset of autoreactive plasma cells is EBV lytically infected in the RA synovia and SS salivary glands; iv) antibodies specific for unmodified and citrullinated EBV peptides, known to cross-recognize ACPA, are produced within ectopic lymphoid structures as 8 demonstrated in vivo in human RA/SCID chimeras; v) SS salivary gland grafts transplanted into SCID mice release human IgG against EBV antigens, whose production correlates with the level of SS-associated auto-antibodies and vi) analysis of CD8+ and CD4+ T-cell localization and granzyme B expression indicated that EBV persistence in ELS-containing RA synovia and SS salivary glands may be favoured by exclusion of CD8+ T cells from B-cell follicles and impaired CD8-mediated cytotoxicity. Overall, these results redefine a novel and pathogenically relevant role for EBV in B-cell dysregulation and chronic inflammation in RA synovium and SS salivary glands

EuPRAXIA@SPARC_LAB: the high-brightness RF photo-injector layout proposal

At EuPRAXIA@SPARC_LAB, the unique combination of an advanced high-brightness RF injector and a plasma-based accelerator will drive a new multi-disciplinary user-facility. The facility, that is currently under study at INFN-LNF Laboratories (Frascati, Italy) in synergy with the EuPRAXIA collaboration, will operate the plasma-based accelerator in the external injection configuration. Since in this configuration the stability and reproducibility of the acceleration process in the plasma stage is strongly influenced by the RF-generated electron beam, the main challenge for the RF injector design is related to generating and handling high quality electron beams. In the last decades of R&D activity, the crucial role of high-brightness RF photo-injectors in the fields of radiation generation and advanced acceleration schemes has been largely established, making them effective candidates to drive plasma-based accelerators as pilots for user facilities. An RF injector consisting in a high-brightness S-band photo-injector followed by an advanced X-band linac has been proposed for the EuPRAXIA@SPARC_LAB project. The electron beam dynamics in the photo-injector has been explored by means of simulations, resulting in high-brightness, ultra-short bunches with up to 3 kA peak current at the entrance of the advanced X-band linac booster. The EuPRAXIA@SPARC_LAB high-brightness photo-injector is described here together with performance optimisation and sensitivity studies aiming to actual check the robustness and reliability of the desired working point.Comment: 5 pages,5 figures, EAAC201

EUPRAXIA@SPARC_LAB: Beam Dynamics studies for the X-band Linac

In the framework of the Eupraxia Design Study an advanced accelerator facility EUPRAXIA@SPARC_LAB has been proposed to be realized at Frascati (Italy) Laboratories of INFN. Two advanced acceleration schemes will be applied, namely an ultimate high gradient 1 GeV X-band linac together with a plasma acceleration stage to provide accelerating gradients of the GeV/m order. A FEL scheme is foreseen to produce X-ray beams within 3-10 nm range. A 500-TW Laser system is also foreseen for electron and ion production experiments and a Compton backscattering Interaction is planned together with extraction beamlines at intermediate electron beam energy for neutron beams and THz radiation production. The electron beam dynamics studies in the linac are here presented together with the preliminary machine layout.Comment: 5 pages, 3 figures, NIM-A proceedings of EAAC201

Nailfold capillaroscopy abnormalities are associated with the presence of anti-endothelial cell antibodies in Sjogren's syndrome

[No abstract available

Electromagnetic and beam dynamics studies for high gradient accelerators at terahertz frequencies

THz radiation is one of the most appealing portion of the electromagnetic spectrum in terms of multi-disciplinary use in basic science and technology. Beyond the numerous applications, a great interest is its potential for future, compact linear accelerators. Conventional radio-frequency accelerating structures operating at the S and C band can reach gradients up to 30 - 50MV/m, respectively; higher accelerating gradients, of the order of 100MV/m, have been obtained with X-band cavities. THz-based accelerating structures enable operation at even higher gradient, potentially up to the GV/m scale, holding great potential for their application to free-electron lasers and linear colliders, for instance. Here we present electromagnetic and beam dynamics studies about the use of a dielectric loaded waveguide to accelerate electron bunches by mean of a narrow-band multi-cycle THz pulse. The excitation of the accelerating structure by the THz pulse and the bunch acceleration in the excited field are investigated through CST Microwave Studio and GPT simulations

Epstein-Barr virus persistence and infection of autoreactive plasma cells in synovial lymphoid structures in rheumatoid arthritis.

OBJECTIVES: Rheumatoid arthritis (RA) is associated with an increased Epstein-Barr virus (EBV) blood DNA load, a robust immune response to EBV and cross-reactive circulating antibodies to viral and self-antigens. However, the role of EBV in RA pathogenesis remains elusive. Here, we investigated the relationship between synovial EBV infection, ectopic lymphoid structures (ELS) and immunity to citrullinated self and EBV proteins. METHODS: Latent and lytic EBV infection was investigated in 43 RA synovial tissues characterised for presence/absence of ELS and in 11 control osteoarthritis synovia using RT-PCR, in situ hybridisation and immunohistochemistry. Synovial production of anti-citrullinated protein (ACPA) and anti-citrullinated EBV peptide (VCP1/VCP2) antibodies was investigated in situ and in vivo in the severe combined immunodeficiency (SCID)/RA chimeric model. RESULTS: EBV dysregulation was observed exclusively in ELS+ RA but not osteoarthritis (OA) synovia, as revealed by presence of EBV latent (LMP2A, EBV-encoded small RNA (EBER)) transcripts, EBER+ cells and immunoreactivity for EBV latent (LMP1, LMP2A) and lytic (BFRF1) antigens in ELS-associated B cells and plasma cells, respectively. Importantly, a large proportion of ACPA-producing plasma cells surrounding synovial germinal centres were infected with EBV. Furthermore, ELS-containing RA synovia transplanted into SCID mice supported production of ACPA and anti-VCP1/VCP2 antibodies. Analysis of CD4+ and CD8+ T-cell localisation and granzyme B expression suggests that EBV persistence in ELS-containing synovia may be favoured by exclusion of CD8+ T cells from B-cell follicles and impaired CD8-mediated cytotoxicity. CONCLUSIONS: We demonstrated active EBV infection within ELS in the RA synovium in association with local differentiation of ACPA-reactive B cells

Longitudinal phase-space manipulation with beam-driven plasma wakefields

The development of compact accelerator facilities providing high-brightness beams is one of the most challenging tasks in field of next-generation compact and cost affordable particle accelerators, to be used in many fields for industrial, medical and research applications. The ability to shape the beam longitudinal phase-space, in particular, plays a key role to achieve high-peak brightness. Here we present a new approach that allows to tune the longitudinal phase-space of a high-brightness beam by means of a plasma wakefields. The electron beam passing through the plasma drives large wakefields that are used to manipulate the time-energy correlation of particles along the beam itself. We experimentally demonstrate that such solution is highly tunable by simply adjusting the density of the plasma and can be used to imprint or remove any correlation onto the beam. This is a fundamental requirement when dealing with largely time-energy correlated beams coming from future plasma accelerators

Temperature analysis in the shock waves regime for gas-filled plasma capillaries in plasma-based accelerators

Plasma confinement represents a crucial point for plasma-based accelerators and plasma lenses because it can strongly affect the beam properties. For this reason, an accurate measurement of the plasma parameters, as plasma temperature, pressure and electron density, must be performed. In this paper, we introduce a novel method to detect the plasma temperature and the pressure for gas-filled capillaries in use at the SPARC-LAB test facility. The proposed method is based on the shock waves produced at the ends of the capillary during the gas discharge and the subsequent plasma formation inside it. By measuring the supersonic speed of the plasma outflow, the thermodynamic parameters have been obtained both outside and inside the capillary. A plasma temperature around 1.4 eV has been measured, that depends on the geometric properties and the operating conditions of the capillary