PASD1: a promising target for the immunotherapy of haematological malignancies

In general, there is a lack of good immunotherapy targets within the spectrum of haematological malignancies. However haematopoietic stem cell transplants and continuing antigen discovery have allowed further insight into how further improvements in outcomes for patients might be achieved. Most patients with haematological malignancies can be treated with conventional therapies such as radio- and chemotherapy and will attain first remission. However the removal of residual diseased cells is essential to prevent relapse and its associated high mortality. PASD1 is one of the most tissue restricted cancer-testis (CT) antigens with expression limited to primary spermatagonia in healthy tissue. However, characterisation of PASD1 expression in cancers has been predominantly focussed on haematological malignancies where the inappropriate expression of PASD1 was first identified. PASD1 has one of the highest frequencies of expression of all CT antigens in acute myeloid leukaemia, with some suggestion of its role as a biomarker in diffuse large B-cell lymphoma. Here we describe the characterisation of the function and expression patterns of PASD1 in cell lines and primary tissues. Development of DNA vaccines targeting PASD1 epitopes demonstrate effective ex vivo T-cell responses in terms of IFNγ secretion and tumour cell killing. Of particular note these vaccines have led to the destruction of cells which process and present endogenous PASD1 indicating that effectively primed CTLs could kill PASD1-positive tumour cells

Crustal Structure and Lithospheric Doming: Aspects of Deformation Along an Obliquely Convergent Plate Margin, New Zealand

Lithospheric deformation along and adjacent to the Pacific-Australian Plate boundary through New Zealand has resulted in different expressions in North and South Islands. This thesis investigates some aspects of crustal and upper mantle structure in New Zealand and is divided into two distinct parts. The first examines the structure of the obliquely compressional crustal plate boundary in South Island using seismic techniques; the second focuses on the domed topography of central North Island and its relationship to mantle processes. High density active source, one and three-component, seismic data from a transect across the Southern Alps provides information on the deformation of the crust across the Australia-Pacific plate boundary of South Island. These data show 0-0.08 s ([approximately] 0.25 %) delay times between the radial and transverse directions for shear waves (Sg and SmS phases), with maximum possible delays of 140 ms and the fast direction aligned with the transverse direction (approximately parallel to the plate boundary). The transect is perpendicular to the Alpine Fault, which is slightly oblique to the fast mantle directions determined from SKS phases. The small values of crustal splitting may result from the oblique angle of the ray paths to the actual crustal structure at depth, or the complex nature of the deformation as observed at the surface, which though on a small scale can be strongly anisotropic, may not add constructively over a large region. Poisson's ratio, determined from forward modelling of both P and S phases, shows low values of 0.21 - 0.24 for the crust of South Island. A broad region of low values ([sigma]=0.15) exists at 10-20 km depth under the Southern Alps, which corresponds to a previously identified body of low Vp and high resistivity. The low [sigma] is interpreted as low pore fluid pressure and high silica composition rocks. This contrasts with previous interpretations of iii iv high pore fluid pressure at this depth. The topography of central North Island, New Zealand, describes a 250 km wide and [approximately] 500 m high dome. Exhumation estimates from mudstone porosity measurement indicate an increase in exhumation from [approximately] 500 m at the coast to 2 km in the region of the present topographic high. Combining these values gives an estimate of rock uplift of over 2.5 km for central North Island, since 4 Ma, a rate of 0.6 mm/yr. Tectonic uplift of 1.25 km indicates that [approximately] 50 % of the rock uplift occurs in response to exhumation. An independent local estimation of differential erosion in central North Island gave 300 m of exhumation since at least 500 ka, a rate of [greater than or equal to] 0.6 mm/yr. Using a digital elevation model of New Zealand the fluvial incision of the landscape was calculated and [approximately]169 m of rebound can be attributed to incision. Contouring maximum incision elucidates a region of high incision [approximately] 50 km south of the present centre of domed rock uplift. Using incision as a proxy for rock uplift, it is hypothesised that the incision signal is recent and demonstrates the southward migration of the centre of rock uplift. Rebound of sedimentary basins due to a reduction in plate coupling forces can also account for some of the observed rock uplift. Buoyancy forces required to create the pattern and magnitude of rock uplift are investigated using a 3 D loading model of the lithosphere. Strong upward forces (65 MPa) are required under the Central Volcanic Region, combined with broad uplift (36 MPa) over western North Island, to fit the observed rock uplift. Low Pn velocities under the Central Volcanic Region indicate temperatures 500 [degrees] C hotter than that of normal mantle. This temperature anomaly corresponds to 60 kg/[cubic metre] less dense than normal mantle, which is consistent with the change in density of 66 kg/[cubic metre] estimated from the loading model and aassuming the density change occurs over a 100 km depth range. The southern extent of buoyancy forces does not correspond well to regions of high seismic attenuation in the lithosphere but instead with the region of high incision

Optimization Of The Hockey Fans In Training (Hockey FIT) Weight Loss And Healthy Lifestyle Program For Male Hockey Fans

BACKGROUND: The health outcomes of men continue to be poorer than women globally. Challenges in addressing this problem include difficulties engaging men in weight loss programs as they tend to view these programs as contrary to the masculine narrative of independence and self-reliance. Researchers have been turning towards sports fans to engage men in health promotion programs as sports fans are typically male, and tend to have poor health habits. METHODS: Developed from the highly successful gender-sensitized Football Fans in Training program, Hockey Fans in Training (Hockey FIT) recruited 80 male hockey fans of the London Knights and Sarnia Sting who were overweight or obese into a weekly, 90-minute classroom education and group exercise program held over 12 weeks; a 40-week minimally-supported phase followed. A process evaluation of the Hockey FIT program was completed alongside a pragmatic randomized controlled trial and outcome evaluation in order to fully explore the acceptability of the Hockey FIT program from the perspectives of coaches delivering and participants engaged in the program. Data sources included attendance records, participant focus groups, coach interviews, assessment of fidelity (program observations and post-session coach reflections), and 12-month participant interviews. RESULTS: Coaches enjoyed delivering the program and found it simple to deliver. Men valued being among others of similar body shape and similar weight loss goals, and found the knowledge they gained through the program helped them to make and maintain health behaviour changes. Suggested improvements include having more hockey-related information and activities, greater flexibility with timing of program delivery, and greater promotion of technology support tools. CONCLUSIONS: We confirmed Hockey FIT was an acceptable gender-sensitized health promotion program for male hockey fans who were overweight or obese. Minor changes were required for optimization, which will be evaluated in a future definitive trial

Application of the pMHC array to characterise tumour antigen specific T cell populations in leukaemia patients at disease diagnosis

Immunotherapy treatments for cancer are becoming increasingly successful, however to further improve our understanding of the T-cell recognition involved in effective responses and to encourage moves towards the development of personalised treatments for leukaemia immunotherapy, precise antigenic targets in individual patients have been identified. Cellular arrays using peptide-MHC (pMHC) tetramers allow the simultaneous detection of different antigen specific T-cell populations naturally circulating in patients and normal donors. We have developed the pMHC array to detect CD8+ T-cell populations in leukaemia patients that recognise epitopes within viral antigens (cytomegalovirus (CMV) and influenza (Flu)) and leukaemia antigens (including Per Arnt Sim domain 1 (PASD1), MelanA, Wilms’ Tumour (WT1) and tyrosinase). We show that the pMHC array is at least as sensitive as flow cytometry and has the potential to rapidly identify more than 40 specific T-cell populations in a small sample of T-cells (0.8–1.4 x 106). Fourteen of the twenty-six acute myeloid leukaemia (AML) patients analysed had T cells that recognised tumour antigen epitopes, and eight of these recognised PASD1 epitopes. Other tumour epitopes recognised were MelanA (n = 3), tyrosinase (n = 3) and WT1126-134 (n = 1). One of the seven acute lymphocytic leukaemia (ALL) patients analysed had T cells that recognised the MUC1950-958 epitope. In the future the pMHC array may be used provide point of care T-cell analyses, predict patient response to conventional therapy and direct personalised immunotherapy for patients

Anti-ALK Antibodies in Patients with ALK-Positive Malignancies Not Expressing NPM-ALK.

Patients with Nucleophosmin (NPM)- Anaplastic Lymphoma Kinase (ALK) fusion positive Anaplastic Large Cell Lymphoma produce autoantibodies against ALK indicative of an immune response against epitopes of the chimeric fusion protein. We asked whether ALK-expression in other malignancies induces specific antibodies. Antibodies against ALK were detected in sera of one of 50 analysed ALK-expressing neuroblastoma patients, 13 of 21 ALK positive non-small cell lung carcinoma (NSCLC) patients, 13 of 22 ALK translocation-positive, but NPM-ALK-negative lymphoma patients and one of one ALK-positive rhabdomyosarcoma patient, but not in 20 healthy adults. These data suggest that boosting a pre-existent anti-ALK immune response may be more feasible for patients with ALK-positive NSCLC, lymphomas and rhabdomyosarcomas than for tumours expressing wild-type ALK.This work was supported by a grant from the Deutsche Jose Carreras Leukämie-Stiftung (DJCLS08/ 09) to WW, and RS. CDW and WW are additionally supported by the von behring röntgen stiftung (60-0011) and the Forschungshilfe Peiper, IO and RS by the Kinderkrebsinitiative Buchholz/HolmSeppensen and KP by Bloodwise, the Julian Starmer-Smith Memorial Fund and the Sam Foye Fund. SDT is a Bloodwise University Senior Lecturer and is also supported by the Alex Hulme Foundation and the Sam Foye Fund. MH is supported by a PhD studentship from the Wellcome Trust.This is the final version of the article. It first appeared from Ivyspring International via http://dx.doi.org/10.7150/jca.1523

Sp17 Protein Expression and Major Histocompatibility Class I and II Epitope Presentation in Diffuse Large B Cell Lymphoma Patients

Improved therapies are urgently needed for patients with diffuse large B cell lymphoma (DLBCL). Success using immune checkpoint inhibitors and chimeric antigen receptor T cell technology has fuelled demand for validated cancer epitopes. Immunogenic cancer testis antigens (CTAs), with their widespread expression in many tumours but highly restricted normal tissue distribution, represent attractive immunotherapeutic targets that may improve treatment options for DLBCL and other malignancies. Sperm protein 17 (Sp17), a CTA reported to be immunogenic in ovarian cancer and myeloma patients, is expressed in DLBCL. The aim of the present study was to investigate Sp17 epitope presentation via the presence of a cytotoxic T cell (CTL) and a CD4 T-helper (Th) response in DLBCL patients. A significant γ-interferon CTL response was detected in peripheral blood mononuclear cells of 13/31 DLBCL patients following short-term cell stimulation with two novel HLA-A⁎0201 peptides and one previously reported HLA-A⁎0101-restricted nine-mer Sp17 peptide. No significant responses were detected in the HLA-A⁎0201-negative DLBCL patients or four healthy subjects. A novel immunogenic 20-mer CD4 Th Sp17 peptide was detected in 8/17 DLBCL patients. This is the first report of a CTL and a CD4 Th response to Sp17 in DLBCL and supports Sp17 as a potential immunotherapeutic target for DLBCL

Identification and Characterization of Peripheral T-Cell Lymphoma-Associated SEREX Antigens

Peripheral T-cell lymphomas (PTCL) are generally less common and pursue a more aggressive clinical course than B-cell lymphomas, with the T-cell phenotype itself being a poor prognostic factor in adult non-Hodgkin lymphoma (NHL). With notable exceptions such as ALK+ anaplastic large cell lymphoma (ALCL, ALK+), the molecular abnormalities in PTCL remain poorly characterised. We had previously identified circulating antibodies to ALK in patients with ALCL, ALK+. Thus, as a strategy to identify potential antigens associated with the pathogenesis of PTCL, not otherwise specified (PTCL, NOS), we screened a testis cDNA library with sera from four PTCL, NOS patients using the SEREX (serological analysis of recombinant cDNA expression libraries) technique. We identified nine PTCL, NOS-associated antigens whose immunological reactivity was further investigated using sera from 52 B- and T-cell lymphoma patients and 17 normal controls. The centrosomal protein CEP250 was specifically recognised by patients sera and showed increased protein expression in cell lines derived from T-cell versus B-cell malignancies. TCEB3, BECN1, and two previously uncharacterised proteins, c14orf93 and ZBTB44, were preferentially recognised by patients' sera. Transcripts for all nine genes were identified in 39 cancer cell lines and the five genes encoding preferentially lymphoma-recognised antigens were widely expressed in normal tissues and mononuclear cell subsets. In summary, this study identifies novel molecules that are immunologically recognised in vivo by patients with PTCL, NOS. Future studies are needed to determine whether these tumor antigens play a role in the pathogenesis of PTCL

Characteristics of Salmonella recovered from stools of children enrolled in the Global Enteric Multicenter Study

Background: The Global Enteric Multicenter Study (GEMS) determined the etiologic agents of moderate-To-severe diarrhea (MSD) in children under 5 years old in Africa and Asia. Here, we describe the prevalence and antimicrobial susceptibility of nontyphoidal Salmonella (NTS) serovars in GEMS and examine the phylogenetics of Salmonella Typhimurium ST313 isolates. Methods: Salmonella isolated from children with MSD or diarrhea-free controls were identified by classical clinical microbiology and serotyped using antisera and/or whole-genome sequence data. We evaluated antimicrobial susceptibility using the Kirby-Bauer disk-diffusion method. Salmonella Typhimurium sequence types were determined using multi-locus sequence typing, and whole-genome sequencing was performed to assess the phylogeny of ST313. Results: Of 370 Salmonella-positive individuals, 190 (51.4%) were MSD cases and 180 (48.6%) were diarrhea-free controls. The most frequent Salmonella serovars identified were Salmonella Typhimurium, serogroup O:8 (C2-C3), serogroup O:6,7 (C1), Salmonella Paratyphi B Java, and serogroup O:4 (B). The prevalence of NTS was low but similar across sites, regardless of age, and was similar among both cases and controls except in Kenya, where Salmonella Typhimurium was more commonly associated with cases than controls. Phylogenetic analysis showed that these Salmonella Typhimurium isolates, all ST313, were highly genetically related to isolates from controls. Generally, Salmonella isolates from Asia were resistant to ciprofloxacin and ceftriaxone, but African isolates were susceptible to these antibiotics. Conclusions: Our data confirm that NTS is prevalent, albeit at low levels, in Africa and South Asia. Our findings provide further evidence that multidrug-resistant Salmonella Typhimurium ST313 can be carried asymptomatically by humans in sub-Saharan Africa

New insights into the genetic etiology of Alzheimer's disease and related dementias

Characterization of the genetic landscape of Alzheimer's disease (AD) and related dementias (ADD) provides a unique opportunity for a better understanding of the associated pathophysiological processes. We performed a two-stage genome-wide association study totaling 111,326 clinically diagnosed/'proxy' AD cases and 677,663 controls. We found 75 risk loci, of which 42 were new at the time of analysis. Pathway enrichment analyses confirmed the involvement of amyloid/tau pathways and highlighted microglia implication. Gene prioritization in the new loci identified 31 genes that were suggestive of new genetically associated processes, including the tumor necrosis factor alpha pathway through the linear ubiquitin chain assembly complex. We also built a new genetic risk score associated with the risk of future AD/dementia or progression from mild cognitive impairment to AD/dementia. The improvement in prediction led to a 1.6- to 1.9-fold increase in AD risk from the lowest to the highest decile, in addition to effects of age and the APOE ε4 allele