Lindsay Nicholson - Academia.edu (original) (raw)

Papers by Lindsay Nicholson

British journal of haematology, Jan 27, 2015

Glucocorticoid (GC) resistance is a continuing clinical problem in childhood acute lymphoblastic ... more Glucocorticoid (GC) resistance is a continuing clinical problem in childhood acute lymphoblastic leukaemia (ALL) but the underlying mechanisms remain unclear. A proteomic approach was used to compare profiles of the B-lineage ALL GC-sensitive cell line, PreB 697, and its GC-resistant sub-line, R3F9, pre- and post-dexamethasone exposure. PAX5, a transcription factor critical to B-cell development was differentially regulated in the PreB 697 compared to the R3F9 cell line in response to GC. PAX5 basal protein expression was less in R3F9 compared to its GC-sensitive parent and confirmed to be lower in other GC-resistant sub-lines of Pre B 697 and was associated with a decreased expression of the PAX5 transcriptional target, CD19. Gene set enrichment analysis showed that increasing GC-resistance was associated with differentiation from preB-II to an immature B-lymphocyte stage. GC-resistant sub-lines were shown to have higher levels of phosphorylated JNK compared to the parent line and ...

The delineation of debris-covered glaciers remains a challenge in optical remote sensing, due to ... more The delineation of debris-covered glaciers remains a challenge in optical remote sensing, due to the similarity of the spectral signature of debris-covered ice to surrounding lateral moraines, making it difficult to apply standard semi-automated algorithms commonly used for clean ice delineation. Furthermore, supraglacial debris exhibits considerable spatial variability in its characteristics such as debris cover thickness, particle size, thermal resistance and thermal conductivity. These properties are needed in order to map the extent of debris cover and to estimate ice melt under the debris cover or at the surface. In this study we evaluate the potential of texture analysis for detecting surface characteristics of debris-cover glacier tongues in the Khumbu Himalaya, using Advanced Spaceborne Thermal Emission and Reflection Radiometer (ASTER) and high-resolution Ikonos data. We focus on mapping supra-glacier lakes and exposed ice walls using texture analysis algorithms such as gre...

Immunology, 2015

Experimental Autoimmune Uveoretinitis (EAU) in the C57BL/6J mouse is a model of non-infectious po... more Experimental Autoimmune Uveoretinitis (EAU) in the C57BL/6J mouse is a model of non-infectious posterior segment intraocular inflammation, which parallels clinical features of the human disease. The purpose of this study was to analyse the immune response to the four murine subunits of Retinol Binding Protein-3 (RBP-3) to identify pathogenic epitopes to investigate the presence of intramolecular epitope spreading during the persistent inflammation phase observed in this model of EAU. Recombinant murine subunits of the RBP-3 protein were purified and used to immunise C57BL/6J mice to induce EAU. An overlapping peptide library was used to screen RBP-3 subunit 3 for immunogenicity and pathogenicity. Disease phenotype and characterisation of pathogenic subunits and peptides was undertaken by topical endoscopic fundal imaging (TEFI), immunohistochemistry, proliferation assays and flow cytometry. RBP-3 subunits 1, 2 and 3 induced EAU in the C57BL/6J mice, with subunit 3 eliciting the most destructive clinical disease. Within subunit 3 we identified a novel uveitogenic epitope, 629-643. The disease induced by this peptide was comparable to that produced by the uveitogenic 1-20 peptide. Following immunisation peptide specific responses by CD4(+) and CD8(+) T cell subsets were detected, and cells from both populations were present in the retinal inflammatory infiltrate. Intramolecular epitope spreading between 629-643 and 1-20 was detected in mice with clinical signs of disease. 629-643 RBP-3 peptide is a major uveitogenic peptide for the induction of EAU in C57BL/6J mice and the persistent clinical disease induced with one peptide leads to epitope spreading. This article is protected by copyright. All rights reserved.

A clear understanding of the events surrounding the selection of autoreactive T cells in the thym... more A clear understanding of the events surrounding the selection of autoreactive T cells in the thymus and their regulation in the periphery has eluded immunologists for years. However, recent work examining the expression of tissue-specific antigens in the thymus and the biochemistry of disease associated MHC alleles has provided important clues into the generation of the autoreactive T cell repertoire

PLOS ONE, 2015

Current clinical trials utilize mesenchymal stromal cells (MSCs) expanded in culture, however the... more Current clinical trials utilize mesenchymal stromal cells (MSCs) expanded in culture, however these interventions carry considerable costs and concerns pertaining to culture-induced losses of potency. This study assessed the feasibility of new clinical-grade technology to obtain uncultured MSC isolates from three human intra-osseous tissue sources based on immunomagnetic selection for CD271-positive cells.

Proximal signaling events activated by TCR-peptide/MHC (TCR-pMHC) binding have been the focus of ... more Proximal signaling events activated by TCR-peptide/MHC (TCR-pMHC) binding have been the focus of intense ongoing study, but understanding how the consequent downstream signaling networks integrate to govern ultimate avidity-appropriate TCR- pMHC T cell responses remains a crucial next challenge. We hypothesized that a quantitative combination of key downstream network signals across multiple pathways must encode the information generated by TCR

Annual review of immunology, 2002

T cells that can respond to self-antigens are present in the peripheral immune repertoire of all ... more T cells that can respond to self-antigens are present in the peripheral immune repertoire of all healthy individuals. Recently we have found that unmanipulated SJL mice that are highly susceptible to EAE also maintain a very high frequency of T cells responding to an encephalitogenic epitope of a myelin antigen proteolipid protein (PLP) 139-151 in the peripheral repertoire. This is not due to lack of expression of myelin antigens in the thymus resulting in escape of PLP 139-151 reactive cells from central tolerance, but is due to expression of a splice variant of PLP named DM20, which lacks the residues 116-150. In spite of this high frequency, the PLP 139-151 reactive cells remain undifferentiated in the periphery and do not induce spontaneous EAE. In contrast, SJL TCR transgenic mice expressing a receptor derived from a pathogenic T cell clone do develop spontaneous disease. This may be because in normal mice, autoreactive cells are kept in check by an alternate PLP 139-151 reacti...

Computerized Medical Imaging and Graphics, 2014

This paper presents novel pre-processing image enhancement algorithms for retinal optical coheren... more This paper presents novel pre-processing image enhancement algorithms for retinal optical coherence tomography (OCT). These images contain a large amount of speckle causing them to be grainy and of very low contrast. To make these images valuable for clinical interpretation, we propose a novel method to remove speckle, while preserving useful information contained in each retinal layer. The process starts with multi-scale despeckling based on a dual-tree complex wavelet transform (DT-CWT). We further enhance the OCT image through a smoothing process that uses a novel adaptiveweighted bilateral filter (AWBF). This offers the desirable property of preserving texture within the OCT image layers. The enhanced OCT image is then segmented to extract inner retinal layers that contain useful information for eye research. Our layer segmentation technique is also performed in the DT-CWT domain. Finally we describe an OCT/fundus image registration algorithm which is helpful when two modalities are used together for diagnosis and for information fusion.

2013 IEEE International Conference on Image Processing, 2013

This paper presents an image enhancement method for retinal optical coherence tomography (OCT) im... more This paper presents an image enhancement method for retinal optical coherence tomography (OCT) images. Raw OCT images contain a large amount of speckle which causes images to be grainy and very low contrast. The raw OCT images are thus difficult usually processed before any clinical interpretation is made. We propose a novel method to remove speckle, while preserving useful information contained in each retinal layer. The process starts with multi-scale despeckling based on a dual-tree complex wavelet transform (DT-CWT). Then, we further enhance the OCT image with a novel smoothing process that uses novel adaptive-weighted bilateral filter (AWBF). This offers desirable property of preserving texture within the OCT images. Glaucoma classification results further confirm that our method can enhance the clinical usefulness of OCT images.

The FASEB Journal, 2006

Glucocorticoids (GCs) specifically induce apoptosis in malignant lymphoblasts and are thus pivota... more Glucocorticoids (GCs) specifically induce apoptosis in malignant lymphoblasts and are thus pivotal in the treatment of acute lymphoblastic leukemia (ALL). However, GC-resistance is a therapeutic problem with an unclear molecular mechanism. We generated ϳ70 GC-resistant sublines from a GC-sensitive B-and a T-ALL cell line and investigated their mechanisms of resistance. In response to GCs, all GC-resistant subclones analyzed by real-time polymerase chain reaction (PCR) showed a deficient up-regulation of the GC-receptor (GR) and its downstream target, GC-induced leucine zipper. This deficiency in GR up-regulation was confirmed by Western blotting and on retroviral overexpression of GR in resistant subclones GC-sensitivity was restored. All GC-resistant subclones were screened for GR mutations using denaturing high-pressure liquid chromatography (DHPLC), DNA-fingerprinting, and fluorescence in situ hybridization (FISH). Among the identified mutations were some previously not associated with GC resistance: A484D, P515H, L756N, Y663H, L680P, and R714W. This approach revealed three genotypes, complete loss of functional GR in the mismatch repair deficient T-ALL model, apparently normal GR genes in B-ALLs, and heterozygosity in both. In the first genotype, deficiency in GR up-regulation was fully explained by mutational events, in the second by a putative regulatory defect, and in the third by a combination thereof. In all instances, GC-resistance occurred at the level of the GR in both models.-Schmidt, S., Irving, J. A. E., Minto, L., Matheson, E., Nicholson, L., Ploner, A., Parson, W., Kofler, A., Amort, M., Erdel, M., Hall, A., Kofler, R. Glucocorticoid resistance in two key models of acute lymphoblastic leukemia occurs at the level of the glucocorticoid receptor. FASEB J. 20, E2087-E2097 (2006)

Progress in Retinal and Eye Research, 2008

Experimental autoimmune uveoretinitis (EAU) serves as an animal model for human uveitis. EAU is i... more Experimental autoimmune uveoretinitis (EAU) serves as an animal model for human uveitis. EAU is inducible in animals by peripheral immunization with proteins found in the retina that triggers an immune response which leads to tissue damage. This is coordinated by autoantigen specific CD4(+) T cells whose activation is accompanied by the infiltration of a wide range of other leukocytes into the retina. Infiltrating macrophages and granulocytes cause destruction by the release of reactive oxygen and nitrogen species but these and other leukocytes also regulate inflammation. This review will describe the dynamics of leukocyte infiltration in EAU from the initial systemic activation of T cells following immunization, through their traffic into the eye causing a peak of infiltration, and ending with a phase of secondary regulation in which, although clinical disease has resolved, the leukocyte composition of the eye remains altered.

Proceedings of the National Academy of Sciences, 2004

SJL mice are highly susceptible to experimental autoimmune encephalomyelitis (EAE) induced with m... more SJL mice are highly susceptible to experimental autoimmune encephalomyelitis (EAE) induced with myelin proteolipid protein (PLP) peptide 139-151, whereas H-2 congenic B10.S mice are resistant. Immunodominance and susceptibility to EAE are associated with a high precursor frequency of PLP 139-151-specific T cells in the naive repertoire of SJL mice. To understand the mechanism of EAE resistance in B10.S mice, we determined the precursor frequency of PLP 139-151-reactive T cells in both strains by using IAs/PLP 139-151 tetramers. SJL and B10.S mice had similar frequencies of tetramer-reactive T cells in the naive peripheral repertoire. However, in SJL mice, the majority of PLP 139-151 tetramer-positive cells were in the CD4+CD25- population, whereas there were more tetramer-positive cells in the CD4+CD25+ population of B10.S mice. Depletion of CD4+CD25+ cells in vivo facilitated the expansion of PLP 139-151-reactive cells with production of T helper 1 cytokines in EAE-resistant B10.S mice. Furthermore, anti-CD25 Ab treatment before immunization resulted in EAE induction in these otherwise resistant mice. These data indicate an important role for autoantigen-specific CD4+CD25+ cells in genetic resistance to autoimmunity.

Molecular Immunology, 2004

T-cells provide antigen specific focus to the adaptive immune response. However, the receptors th... more T-cells provide antigen specific focus to the adaptive immune response. However, the receptors that initiate functional responses are not in themselves specific, as they can recognise many different ligands. In this paper, we propose that the immune system deals with TCR degeneracy by an adaptive process that effects both T-cell activation and T-cell function, and exploits TCR degeneracy to limit pathogenic autoimmune disease.

Methods, 2006

Delivering soluble (auto) antigenic peptides via the naso-respiratory route induces tolerance to ... more Delivering soluble (auto) antigenic peptides via the naso-respiratory route induces tolerance to that peptide and suppression of experimental models of autoimmune disease. In the normal lung, respiratory tract dendritic cells (RTDCs) efficiently endocytose soluble antigens, migrate to regional lymph nodes and present peptide to T cells that subsequently become tolerant. This article describes protocols for inducing tolerance via the naso-respiratory tract in experimental autoimmune uveoretinitis (EAU); for the isolation of RTDCs to facilitate definition of, and conditions for, maturation and activation of cells; and to test RTDC ability to induce tolerance in murine EAU when adoptively transferred.

Leukemia Research, 2010

Glucocorticoids (GCs) are pivotal agents in the treatment of childhood acute lymphoblastic leukae... more Glucocorticoids (GCs) are pivotal agents in the treatment of childhood acute lymphoblastic leukaemia (ALL) but the molecular basis of GC-resistance remains unclear. Expression-array studies have shown that commonly upregulated genes associated with GC-sensitivity include GR, glucocorticoid-induced leucine zipper (GILZ) and IkappaBalpha, which all negatively interact with components of the pro-survival NFkappaB pathway and therefore may be critical determinants of GC-sensitivity. We have investigated these regulators and their effect on NFkappaB activity in GC-resistant descendents of the B-lineage ALL cell line, PreB 697. We show that while differential up regulation of the modulators (GILZ, GR and IkappaBalpha) was demonstrated in GC-sensitive compared to GC-resistant sub-lines, this was not coupled with altered nuclear translocation or functionality of the RelA, p50 or c-Rel subunits of NFkappaB. Thus, GC-resistance in the PreB 697 cell line model is not mediated by NFkappaB, however further investigation of the impact of these GC-sensitive associated proteins on other survival pathways, such as the RAS-RAF-MEK-ERK pathway, is warranted.

Journal of Neuroimmunology, 1998

Cross-reactivity with environmental antigens has been postulated as a mechanism responsible for t... more Cross-reactivity with environmental antigens has been postulated as a mechanism responsible for the induction of autoimmune disease. Experimental autoimmune encephalomyelitis is a T cell-mediated autoimmune disease model inducible in susceptible strains of laboratory animals by immunization with protein constituents of myelin. We used myelin proteolipid protein (PLP) peptide 139-151 and its analogues to define motifs to search a protein database for structural homologues of PLP139-151 and identified five peptides derived from microbial Ags that elicit immune responses that cross-react with this self peptide. Exposure of naive SJL mice to the cross-reactive environmental peptides alone was insufficient to induce autoimmune disease even when animals were treated with Ag-nonspecific stimuli (superantigen or LPS). However, immunization of SJL mice with suboptimal doses of PLP139-151 after priming with cross-reactive environmental peptides consistently induced experimental autoimmune encephalomyelitis. Furthermore, T cell lines from mice immunized with cross-reactive environmental peptides and restimulated in vitro with PLP139-151 could induce disease upon transfer into naive recipients. These data suggest that expansion by self Ag is required to break the threshold to autoimmune disease in animals primed with cross-reactive peptides.

Journal of Molecular Endocrinology, 1996

We have characterized four murine monoclonal antibodies (mAbs) to the extracellular domain of the... more We have characterized four murine monoclonal antibodies (mAbs) to the extracellular domain of the human TSH receptor (TSH-R.E), the target autoantigen of Graves' disease. Recombinant TSH-R.E used as immunogen, was produced in E. coli as a fusion protein with glutathione-S-transferase or in a baculovirus-insect cell system, as a non-fusion glycoprotein. To increase the epitope specificity of the mAbs, two different strains of mice (H-2(b) and H-2(d)) were immunized. The epitopes recognized by the mAbs were characterized by immunoblotting with various recombinant constructs of TSH-R.E and by binding to overlapping synthetic peptides of the receptor. The four IgG mAbs characterized recognized epitopes localized to different regions on the TSH-R.E; amino acids 22-35 (A1O and A11, both IgG2b from H-2(b) animals), amino acids 402-415 (A7, IgG2b from H-2(b) animals) and amino acids 147-228 (A9, IgG1 from H-2(d) animals). Immunolocalization studies showed that mAb A9 recognized TSH-R.E on unfixed cryostat sections, where binding was localized to the basolateral plasma membrane of thyroid follicular cells, suggesting that this antibody reacts with the native receptor on thyroid cells. The binding of the mAbs A7, A10 and A11 was also restricted to the basal surface of thyroid cells, but only after acetone fixation of the sections, implying that the epitopes recognized on the amino and carboxyl terminus of the extracellular region of the receptor are not accessible on the native molecule. None of the mAbs stimulated cyclic AMP responses in COS-7 cells transiently transfected with full-length functioning TSH-R.E, whilst weak inhibition of binding of radiolabelled TSH to porcine membranes in a radioreceptor assay was apparent with mAb A10 and A11, but only at high concentrations of IgG. The ability of mAb A9 to bind to the native receptor without stimulating activity or inhibition of TSH binding suggests that antibody can bind to the central region of the TSH-R.E without perturbing receptor function. The availability of mAbs that recognize epitopes on different regions of the extracellular domain of TSH-R will lead to a better understanding of the autoantigenic regions on TSH-R implicated in disease activity.

The Journal of Immunology, 2005

IFN-gamma stimulates macrophage activation and NO production, which leads to destruction of the r... more IFN-gamma stimulates macrophage activation and NO production, which leads to destruction of the retina in experimental autoimmune uveoretinitis. In this study, we investigate the mechanism of disease resistance in TNF p55 receptor-deficient animals. We show that although T cell priming is relatively unaffected, macrophages lacking the TNF p55 receptor fail to produce NO following IFN-gamma stimulation because of a requirement for autocrine TNF-alpha signaling through the TNF p55 receptor. In contrast to the impaired activation of NO synthesis, MHC class II up-regulation was indistinguishable in wild-type and TNFRp55-/- mice stimulated with IFN-gamma. These defects could be overcome by stimulating macrophages with LPS. Together, these results show that selected aspects of IFN-gamma activation are controlled by autocrine secretion of TNF-alpha, but that this control is lost in the presence of signals generated by pathogen-associated molecular patterns recognizing receptors.