20 research outputs found
Serologic detection of diffuse large B-cell lymphoma-associated antigens
No full textDiffuse large B-cell lymphoma (DLBCL) accounts for 30-40% of all adult non-Hodgkin's lymphomas, yet the understanding of its underlying genetic abnormalities remains poor. Our present study used the serological analysis of recombinant cDNA expression libraries (SEREX) technique to identify DLBCL-associated antigens. SEREX screening of testis libraries has previously identified cancer-testis antigens (CTAs) that may act as disease-specific targets for immunotherapy. Screening a testis cDNA expression library with serum from a DLBCL patient identified a total of 94 positive clones, representing 28 distinct antigens. Two of these antigens were novel, 8 were previously uncharacterised, and the remainder were proteins of known function. Screening of the antigens with sera from DLBCL (n = 10), acute myeloid leukaemia (AML, n = 10) and chronic myeloid leukaemia (CML, n = 10) patients, alongside normal healthy donor controls (n = 20), revealed that 7 of the antigens were recognised by DLBCL sera but not normal donor sera, whilst 2 of these antigens were also recognised by leukaemic sera. Some of the genes identified here were already known to be transcribed in DLBCL. The mRNA expression of the majority of the remaining antigens was confirmed in DLBCL cell lines using reverse-transcriptase PCR (RT-PCR). Our study identified a number of DLBCL associated antigens that may be suitable as prognostic/diagnostic markers and/or for the immunotherapy of haematologic malignancie
Portraits: Creative Spaces
No full textPortraits: Creative Spaces is two series of photographic portraits of artists in their creative environments, produced for the book Making Great Illustration (AC Black, 2011), and subsequently exhibited at the International Festival of Literature, Plymouth, and the University Gallery, Nanjing, China (both 2011). Liggins was invited by Jo Davies, the book’s author, to make photographic portraits of artists in their working environments. Liggins she used the project to develop two contrasting series: (1) detailed subject-in-their-environment photographs achieved digitally from a conventional photographic viewpoint, and (2) development of her baroque photographic aesthetic using low-tech plastic camera technology. The value of the contrast was that it allowed Liggins to identify and accentuate the differences between the two photographic modes. Whereas the conventional perspective with high-resolution detail created a picture of illustrator and environment that is complete and that can be mined for detail, the lack of sophistication in the plastic camera meant that the image was governed by properties arising from its limitations. Obscurity and disruption were introduced, such as the darkening in the corners, and the beginnings of light dispersion as the image is stretched and blurred towards the border of the photographs, that led to a sense of the image being its own world. This contrast prompted another: the high-resolution, conventional perspective produced an objectifying gaze, while the low-tech image suggested a passing glance and evoked a sense of place through the disruptions that draw the viewer in. For examples, see the portraits on Making Great Illustration, p. 214. Her baroque portrait of artist Bedwyr Williams also accompanies the interview with Williams in volume 1 of the journal Cultural Colony Quarterly. Although Davies’s book privileges Liggins’s conventional-perspective images, the accompanying exhibitions allowed the two series to be shown side-by-side in order for the contrast to be displayed fully
Humoral detection of leukaemia-associated antigens in presentation acute myeloid leukaemia
No full textThe serological analysis of recombinant cDNA expression libraries (SEREX) technique was used to immunoscreen a testes cDNA expression library with sera from newly diagnosed acute myeloid leukaemia (AML) patients. We used a testis cDNA library to aid our identification of cancer-testis (CT) antigens. We identified 44 antigens which we further immunoscreened with sera from AML, chronic myeloid leukaemia (CML), and normal donors. Eight antigens were solely recognised by patient sera including the recently described CT antigen, PASD1, and the cancer-related SSX2 interacting protein, SSX2IP. RT-PCR analysis indicated that we had identified three antigens which were expressed in patient bone marrow (BM) and peripheral blood (PB) but not in normal donor samples (PASD1, SSX2IP, and GRINL1A). Real-time PCR (RQ-PCR) confirmed the restricted expression of PASD1 in normal donor organs. Antigen presentation assays using monocyte-derived dendritic cells (mo-DCs) showed that PASD1 could stimulate autologous T-cell responses, suggesting that PASD1 could be a promising target for future immunotherapy clinical trial
Cloning and characterisation of genes encoding molecular recognition proteins from insects
No full textOlfaction is one of the most important senses by which insects obtain information about their environment. In the early stages of olfactory perception in insects, odour molecules are carried across the sensillum lymph by small soluble Odorant Binding Proteins (OBPs). This is followed by activation of the appropriate olfactory receptor, resulting in an electrical impulse, and subsequent degradation of the initial signal.
OBPs have been studied in a range of insect orders including Lepidoptera, Diptera and Orthoptera, and this study reports the cloning and characterisation of cDNAs with a potential olfactory role in the vetch aphid, Megoura viciae (Buckton, Homoptera: Aphididiae).
Construction and sequencing of antennal cDNA libraries identified two cDNAs, MvicOBP1 and Mv164, which were approximately 0.8kb and 1kb respectively. The amino acid sequence of MvicOBP1 has the spacing pattern of six cysteine residues that is characteristic of insect OBPs, and Mv164 shows similarity to insect cytochrome P450 enzymes. RT-PCR showed that these cDNAs have specific or enhanced expression in the chemosensory tissues of M. viciae, and parallel expression patterns suggest a "linked" function. Related sequences are present and expressed in other aphid species, and sequencing of genomic fragments allowed the partial elucidation of the intron/exon organisation of these genes.
Subtracted antennal cDNA libraries identified two cDNAs encoding proteins with significant similarity to insect chemosensory proteins (CSPs), cDNAs encoding Juvenile Hormone Binding Proteins (JHBPs), and a tissue-specific cDNA with a potential carrier role. These, coupled with the OBPs, add evidence to the suggestion that there is an insect superfamily of binding proteins.
A PBP from Bombyx mori (BmorPBP1) was used as a model system for in vitro expression of an insect OBP and subsequent characterisation of the recombinant protein. Four forms of this protein, identified through their interaction with an anti-BmorPBP antibody, were resolved by isoelectric focusing
Identification and characterization of peripheral T-cell lymphoma-associated SEREX antigens.
No full textPeripheral 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
CD4-positive T-helper cell responses to the PASD1 protein in patients with diffuse large B-cell lymphoma
No full textBackground Vaccine development targeting the novel immunogenic Per ARNT Sim Domain containing 1 (PASD1) cancer testis antigen represents an attractive therapeutic approach for the significant number of patients with diffuse large B-cell lymphoma who are refractory to conventional treatment. Since CD4-positive T helper cells have crucial roles in promoting and maintaining immune responses to tumor antigens, the presence of a CD4-positive T-helper immune response to the PASD1 antigen in patients with diffuse large B-cell lymphoma was investigated in the current study.Design and Methods Thirty-one patients with diffuse large B-cell lymphoma (25 with de novo, five with transformed and one with T-cell-rich B-cell lymphoma) were studied. Five immunogenic PASD1 peptides predicted to bind to several major histocompatibiliy complex, class II DR beta 1 alleles were identified using web-based algorithms. Peripheral blood mononuclear cells from patients were used to investigate the immunogenicity of these DR beta 1-restricted peptides in vitro using both gamma-interferon release enzyme-linked immunospot and cytolytic assays.Results Two of the five PASD1 peptides, PASD1(6) and PASD1(7), were shown to be immunogenic in 14 out of 32 patients studied in a gamma-interferon release assay. CD4-positive T-helper cell lines from two patients raised against PASD1 peptides were able to lyse cell lines derived from hematologic malignancies expressing endogenous PASD1 protein.Conclusions This is the first report of a CD4-positive T-helper response to the PASD1 protein in patients with lymphoma. The immunogenic peptides described here represent valuable additional candidates for inclusion in a vaccine to treat patients with PASD1-positive diffuse large B-cell lymphoma whose disease is refractory to conventional therapies
Cytolytic T-cell response to the PASD1 cancer testis antigenin patients with diffuse large B-cell lymphoma
No full textThe identification of immunogenic cancer testis antigens (CTAs) as immunotherapeutic targets represents one approach to improve treatment options for diffuse large B‐cell lymphoma (DLBCL). We previously identified PASD1 [PAS (Per ARNT Sim) domain containing 1 (PASD1)], a DLBCL‐associated CTA that was expressed in a range of hematopoietic malignancies. The aim of the present study was to investigate the presence of a cytotoxic T‐cell (CTL) response to PASD1 in DLBCL patients. A significant γ‐interferon (IFN) release was detected in 21/29 HLA‐A*0201‐positive DLBCL patients (18 de novo DLBCL, two transformed DLBCL and one T‐cell rich B‐cell lymphoma) following short‐term culture of their peripheral blood mononuclear cells stimulated with five HLA‐A*0201‐restricted PASD1 peptides. No significant responses were detected in 21 HLA‐A*0201‐negative DLBCL patients (12 de novo DLBCL, seven transformed DLBCL, two T‐cell rich B‐cell lymphoma) or six normal subjects. CTL cell lines were able to lyse PASD1‐positive tumour cells in a major histocompatibility complex‐Class I dependent manner. The presence of a γ‐IFN response correlated with PASD1 protein expression in the tumour cells in 12/15 cases studied. This is the first report of a CTL response to a CTA in DLBCL. Our results provide additional valuable evidence supporting PASD1 as a potential immunotherapeutic target for the treatment of DLBCL and other malignancies
Deadly diving? Physiological and behavioural management of decompression stress in diving mammals
No full text© The Author(s), 2011. This article is distributed under the terms of the Creative Commons Attribution License. The definitive version was published in Proceedings of the Royal Society B Biological Sciences 279 (2012): 1041-1050, doi:10.1098/rspb.2011.2088.Decompression sickness (DCS; ‘the bends’) is a disease associated with gas uptake at pressure. The basic pathology and cause are relatively well known to human divers. Breath-hold diving marine mammals were thought to be relatively immune to DCS owing to multiple anatomical, physiological and behavioural adaptations that reduce nitrogen gas (N2) loading during dives. However, recent observations have shown that gas bubbles may form and tissue injury may occur in marine mammals under certain circumstances. Gas kinetic models based on measured time-depth profiles further suggest the potential occurrence of high blood and tissue N2 tensions. We review evidence for gas-bubble incidence in marine mammal tissues and discuss the theory behind gas loading and bubble formation. We suggest that diving mammals vary their physiological responses according to multiple stressors, and that the perspective on marine mammal diving physiology should change from simply minimizing N2 loading to management of the N2 load. This suggests several avenues for further study, ranging from the effects of gas bubbles at molecular, cellular and organ function levels, to comparative studies relating the presence/absence of gas bubbles to diving behaviour. Technological advances in imaging and remote instrumentation are likely to advance this field in coming years.This paper and the workshop it stemmed from
were funded by the Woods Hole Oceanographic Institution
Marine Mammal Centre
