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Broad spectrum antibodies to self-antigens and cytokines in RAG deficiency

ABSTRACT: Overall, this work describes the largest cohort of patients with RAG mutations and an associated phenotype consisting of combined immunodeficiency and granulomatous lesions and/or autoimmunity (CID-G/AI). By using multiple methods (microarray, ELISA and multiplex bead technology), we have consistently identified a distinctive signature of anti-cytokine antibodies in patients with RAG-dependent immunodeficiencies, especially in those with CID-G/AI and a history of severe viral infections. These autoantibodies were not detected in a large panel of patients with other forms of primary immunodeficiency, and may therefore represent a novel biomarker panel of this condition. Known autoantigens, cytokines, chemokines, growth factors and receptors were printed onto microarrays. All targets were printed in triplicate. Arrays were probed with diluted plasma and autoantibodies were detected with a AF647 conjugated anti-human IgG secondary.

ORGANISM(S): Homo sapiens

SUBMITTER: Jacob Rosenberg

PROVIDER: E-GEOD-70618 | biostudies-arrayexpress |

REPOSITORIES: biostudies-arrayexpress

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Broad spectrum antibodies to self-antigens and cytokines in RAG deficiency

Project description:Overall, this work describes the largest cohort of patients with RAG mutations and an associated phenotype consisting of combined immunodeficiency and granulomatous lesions and/or autoimmunity (CID-G/AI). By using multiple methods (microarray, ELISA and multiplex bead technology), we have consistently identified a distinctive signature of anti-cytokine antibodies in patients with RAG-dependent immunodeficiencies, especially in those with CID-G/AI and a history of severe viral infections. These autoantibodies were not detected in a large panel of patients with other forms of primary immunodeficiency, and may therefore represent a novel biomarker panel of this condition.

2015-07-08 | GSE70618 | GEO

Nucleic Acid Programmable Protein Arrays (NAPPA) Somatic p53 Mutations

Project description:Purpose: Mutations in TP53 induce autoantibody immune responses in a subset of cancer patients, which have been proposed as biomarkers for early detection. Here, we investigate the association of p53 specific autoantibodies with multiple tumor subtypes and determine the association with p53 mutation status and epitope specificity. Experimental Design: IgG p53 autoantibodies (p53-AAb), were quantified in 412 serum saples using a programmable ELISA assay from patients with serous ovarian, pancreatic adenocarcinoma, and breast cancer. To determine if patients generated mutation specific autoantibodies we designed a panel of the most relevant 51 p53 point mutant proteins, to be displayed on custom programmable protein microarrays. To determine the epitope specificity we displayed 12 overlapping tiling fragments and 38 N- and C-terminal deletions spanning the length of the wild-type p53 proteins. Results: We detected p53-AAb with sensitivities of 58.8% (ovarian), 22% (pancreatic), 32% (triple negative breast cancer), and 10.2% (HER2+ breast cancer) at 94% specificity. Sera with p53-AAb contained broadly-reactive autoantibodies to 51 displayed p53 mutant proteins, demonstrating a polyclonal response to common epitopes. All p53-AAb displayed broad polyclonal immune response to both continuous and discontinuous epitopes at the N- and C-terminus as well as the DNA binding domain. Conclusion and clinical relevance: In this comprehensive analysis, mutations in tumor p53 induce strong, polyclonal autoantibodies with broadly reactive epitope specificity. The immunoreactivity was compared between 60 pancreactic ductal adenocarcinoma cases and 63 benign pancreatic disease controls against 52 unique mutant p53 and 379 human proteins that were printed on microscope slides. [Contributor] Arizona State University

2016-06-01 | E-GEOD-77949 | biostudies-arrayexpress

Autoimmune antigen and serum factor antigen reactivity in a cohort of patients with mycobacterium infection shown previously to be positive and negative for IgG autoantibodies to IFN-gamma.

Project description:The aim of the study was to test wether our microarray platform is capable of detecting previously described IgG autoantibodies to IFN-gamma. We diluted patient sera 1:250 and incubated dilutions on a nitrocellulose-platform array printed with whole protein antigens. In this study, serum from 10 individuals (subsetted into two groups: individuals positive or negative for previously described IFN-gamma-targeted autoantibodies) was profiled for IgG autoantibody reactivity to whole protein autoimmune and serum factor antigens. We found that IFN-gamma-targeted IgG autoantibodies were recognized in the IFNg-pos samples and also identified other array targets significantly associated with anti-IFNg positive individials using the significance analysis of microarrays (SAM) algorithm.

2013-09-24 | E-GEOD-51102 | biostudies-arrayexpress

Plasma autoantibody associated with basal-like breast cancer-GST2 Array

Project description:Background: Basal-like breast cancer (BLBC) is a rare aggressive subtype that is less likely to be detected through mammographic screening. Identification of circulating markers associated with BLBC could have promise in detection and management of this deadly disease. Methods: Using samples from the Polish Breast Cancer study, a high-quality population-based case-control study of breast cancer, we screened 10,000 antigens on protein arrays using 45 BLBC patients and 45 controls, and identified 748 promising plasma autoantibodies (AAbs) associated with BLBC. ELISA assays of promising markers were performed on a total of 145 BLBC cases and 145 age-matched controls. Sensitivities at 98% specificity were calculated and a BLBC classifier was constructed. Results: We identified a 13-AAbs (CTAG1B, CTAG2, TP53, RNF216, PPHLN1, PIP4K2C, ZBTB16, TAS2R8, WBP2NL, DOK2, PSRC1, MN1, TRIM21) that distinguished BLBC from controls with 33% sensitivity and 98% specificity. We also discovered a strong association of TP53 AAb with its protein expression (p=0.009) in BLBC patients. Conclusions: These AAbs warrant further investigation in clinical studies to determine their value for further understanding the biology of BLBC and possible detection. The immunoreactivity was compared between 45 BLBC cases and 45 controls against 10,000 human proteins that printed on microscopic slides

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Influenza peptide reactivity within a cohort of age-stratified individuals pre and post vaccination with the 2008/2009 trivalent influenza vaccine

Project description:The aim of the study was to test the hypothesis that peptide array reactivity will reveal patterns that can be associated with poor or effective neutralization response to seasonal influenza vaccine We diluted patient sera 1:80 and incubated dilutions on a streptavidin-coated glass surface that was printed with biotinylated peptides derived from seasonal strains of influenza hemagglutinin included in the 2008/2009 trivalent influenza vaccine In this study, serum from 76 study participants, subsetted into young (?30 years old) and elderly (?60 years old) categories were profiled for IgG/IgM antibody reactivity to influenza hemagglutinin peptudes. Using multivariate linear regression and correcting for multiple hypothesis testing by cross-validation, ee found that patterns of peptide reactivity associated with a poor or effective neutralization response to H1N1 and B strains of influenza.

2013-05-01 | E-GEOD-43446 | biostudies-arrayexpress

Autoimmune antigen and serum factor antigen reactivity in a cohort of patients with autoimmune polyendocrinopathy syndrome type 1(APS-1) and healthy controls.

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