The SPARX Project: R&D Activity towards X-rays FEL Sources

SPARX is an evolutionary project proposed by a collaboration among ENEA-INFN-CNR-Università di Roma Tor Vergata aiming at the construction of a FEL-SASE X-ray source in the Tor Vergata Campus. The first phase of the SPARX project, funded by Government Agencies, will be focused on the R&D activity on critical components and techniques for future X-ray facilities. The R&D plans for the FEL source will be developped along two lines: (a) use of the SPARC high brightness photo-injector to develop experimental test on RF compression techniques and other beam physics issues, like emittance degradation in magnetic compressors due to CSR; (b) development of new undulator design concepts and up-grading of the FEL SPARC source to enhance the non linear harmonic generation mechanism, design and test of e-beam conditioning, prebunching and seeding. A parallel program will be aimed at the development of high repetition rate S-band gun, high Quantum Efficiency cathodes, high gradient X-band RF accelerating structures and harmonic generation in gas. In a second phase we plan to explore production of X-rays in a SASE-FEL with harmonic generation, upgrading existing facilities

Characterization of Specific Immune Responses to Different Aspergillus Antigens during the Course of Invasive Aspergillosis in Hematologic Patients

Several studies in mouse model of invasive aspergillosis (IA) and in healthy donors have shown that different Aspergillus antigens may stimulate different adaptive immune responses. However, the occurrence of Aspergillus-specific T cells have not yet been reported in patients with the disease. In patients with IA, we have investigated during the infection: a) whether and how specific T-cell responses to different Aspergillus antigens occur and develop; b) which antigens elicit the highest frequencies of protective immune responses and, c) whether such protective T cells could be expanded ex-vivo. Forty hematologic patients have been studied, including 22 patients with IA and 18 controls. Specific T cells producing IL-10, IFN-\u3b3, IL-4 and IL-17A have been characterized through enzyme linked immunospot and cytokine secretion assays on 88 peripheral blood (PB) samples, by using the following recombinant antigens: GEL1p, CRF1p, PEP1p, SOD1p, \u3b11-3glucan, \u3b21-3glucan, galactomannan. Specific T cells were expanded through short term culture. Aspergillus-specific T cells producing non-protective interleukin-10 (IL-10) and protective interferon-gamma (IFN-\u3b3) have been detected to all the antigens only in IA patients. Lower numbers of specific T cells producing IL-4 and IL-17A have also been shown. Protective T cells targeted predominantly Aspergillus cell wall antigens, tended to increase during the IA course and to be associated with a better clinical outcome. Aspergillus-specific T cells could be successfully generated from the PB of 8 out of 8 patients with IA and included cytotoxic subsets able to lyse Aspergillus hyphae. Aspergillus specific T-cell responses contribute to the clearance of the pathogen in immunosuppressed patients with IA and Aspergillus cell wall antigens are those mainly targeted by protective immune responses. Cytotoxic specific T cells can be expanded from immunosuppressed patients even during the infection by using the above mentioned antigens. These findings may be exploited for immunotherapeutic purposes in patients with I

Cytokine production profile and lytic activities of <i>Aspergillus-</i>specific T cells.

<p><b>A, B.</b> The frequencies of <i>Aspergillus</i> specific T cells producing IFNγ, IL-10, IL-4 or IL-17A against the seven recombinant antigens, either as EM (dark gray portion of the columns) or CM (light gray portion of the columns), are shown as mean % positive cells ± standard deviation, computed over the 22 patients with IA. Results are expressed as percentages of either CD4⁺ T cells (A) or of CD8⁺ T cells (B). <b>C.</b> White columns represent rates of hyphal damage by <i>Aspergillus</i>-specific T cells expanded from PBMCs stimulated with heat killed <i>Aspergillus</i> conidia. Black columns represent rates of hyphal damage by <i>Aspergillus</i>-specific T cells expanded from PBMCs stimulated with <i>Aspergillus</i> recombinant antigens (GEL1, PEP1, α1–3 glucan, β1–3 glucan) at two and twenty two-hour cultures. E/T = effector/target cell ratio. <b>D.</b> Rates of hyphal damage by <i>Aspergillus</i>-specific T cells expanded with <i>Aspergillus</i> either heat killed conidia (white columns) or recombinant antigens (GEL1, PEP1, α1–3 glucan, β1–3 glucan) (black columns); by the supernatant of cultures of <i>Aspergillus</i>-specific T cell lines only; by antigen presenting cells (APCs); by polymophonuclears (PMNs) and by combinations of the different three cell fractions from three further patients with IA, at two-hour cultures. * = <i>P</i><.05. E/T = effector/target cell ratio. Results are expressed as mean+−Standard Deviation.</p

<i>Aspergillus</i>-specific T-cell responses to the 7 recombinant antigens during the course of Invasive Aspegillosis (IA).

<p><b>A,B,C,D. A.</b> Box plots showing specific immune responses producing IL-10 (yellow columns) to any single recombinant antigen. The horizontal axis represents the antigen, which the specific immune responses are directed to. <b>B.</b> Box plot showing specific immune responses producing IL-10 (yellow columns) against all the 7 recombinant antigens at the four phases of the IA. The horizontal axis represents the different phases of IA. <b>C.</b> Box plot showing specific immune responses producing IFN-γ (blue columns) to any single recombinant antigen. The horizontal axis represents the antigen, which the specific immune responses are directed to. <b>D.</b> Box plot showing specific immune responses producing IFN-γ (blue columns) against all the 7 recombinant antigens at the four phases of the IA. The horizontal axis represents the different phases of IA. The vertical axis shows the number of spot-forming cells (SFCs) per million peripheral blood mononuclear cells (PBMCs).The upper horizontal line represents the upper adjacent value. The upper hinge of the boxes represents the 75^th percentile. The middle horizontal line of the boxes represents the median value. The lower hinge of the boxes represents the 25^th percentile. The lower horizontal line represents the lower adjacent value. Yellow dots and blue dots are outrange values.</p

Kinetics of specific T-cell responses to the seven recombinant antigens of <i>Aspergillus</i> by IFN-γ, IL-10, and IL-4 ELISpot assay in 22 patients with invasive aspergillosis (IA), patient 11 to patient 22.

<p>Blue columns represent the number of <i>Aspergillus</i>-specific T cells producing IFN-γ; yellow columns represent the number of <i>Aspergillus</i>-specific T cells producing IL-10; green columns represent the number of <i>Aspergillus</i>-specific T cells producing IL-17A; red columns represent the number of <i>Aspergillus</i>-specific T cells producing IL-4; dark gray background represents T-cell responses in wells with anti-CD3 antibody. Vertical axis shows the number of spot-forming cells (SFCs) per million peripheral blood mononuclear cells (PBMCs); horizontal axis indicates the antigen, which the specific immune responses are directed to and the phase of IA calculated as the number of days elapsed from the radiological diagnosis of the infection (t1, from the radiologic disclosure of the infectious lesions until day +14; t2, from day +15 until day +30; t3, from day +31 to day +45; t4, from day +46 to day 60 days; t4+ >61 days). CRF1 =  ortholog of Crh1p associated in β1,6 glucan-chitin linkages; PEP1 =  aspartic protease; GEL1 = 1,3-β glucanosyltransferase; α1–3 glu = α1–3 glucan; β1–3 glu = β1–3 glucan; SOD1 =  superoxide dismutase; and GM = galactomannan. Pt = patient. * = patients with unfavorable outcome. ^∧ = patients with probable IA. The red dotted line divides the two groups of patients: from pt n° 1 to pt n° 15 with <i>Aspergillus</i>-specific T cells producing IFN-γ to two or more antigens; from pt n° 16 to pt n° 22 without <i>Aspergillus</i>-specific T cells producing IFN-γ or with such cells to only one antigen.</p

Clinical characteristics of the patients.

<p>AML = acute myeloid leukemia; ALL = acute lymphoblastic leukemia; CLL = chronic lymphocytic leukemia; AlloSCT = allogeneic stem cell transplant; AutoSCT = autologous stem cell transplant; DLCBL = diffuse large B cell lymphoma; NHL = non Hodgkin lymphoma; MDS = myelodysplastic syndrome; CNS = central nervous system; IA = proven Invasive Aspergillosis; RSV = respiratory syncytial virus; CMV = cytomegalovirus; E. coli = Escherichia coli; pos = positive; neg = negative; L-AmB = liposomal amphotericin B.</p

Kinetics of specific T-cell responses to the seven recombinant antigens of <i>Aspergillus</i> by IFN-γ, IL-10, and IL-4 ELISpot assay in 22 patients with invasive aspergillosis (IA), patient 1 to patient 10.

<p>Kinetics of specific T-cell responses to the seven recombinant antigens of <i>Aspergillus</i> by IFN-γ, IL-10, and IL-4 ELISpot assay in 22 patients with invasive aspergillosis (IA), patient 1 to patient 10.</p