Protein Microarrays: A New Tool for the Study of Autoantibodies in Immunodeficiency

Rosenberg, Jacob M.; Utz, Paul J.

doi:10.3389/fimmu.2015.00138

Cited by 29 publications

(24 citation statements)

References 77 publications

Supporting

Mentioning

Contrasting

Order By: Relevance

“…Protein microarrays are a powerful tool for the discovery of novel ACAAs in patients with immunodeficiency and autoimmunity and can be applied to any disease context in which serum is available. 17 Additionally, ACAAs have been described in healthy subjects, and protein microarrays provide the opportunity to study how ACAAs can regulate immune homeostasis by using a proteomic approach.…”

Section: Discussionmentioning

confidence: 99%

“…New proteomic tools are needed to query for autoantibodies against hundreds of cytokines, chemokines, and growth factors in health and disease. 17,18 Previously, our laboratory has developed protein microarrays for the detection of ACAAs in patients with the autoimmune disease systemic lupus erythematosus and identified enrichment of ACAAs against B cell–activating factor in patients. 19 We also showed that arrays could detect ACAAs against IFN-α in patients with APS-1 and ACAAs against IFN-γ in patients with atypical mycobacterial infection.…”

mentioning

confidence: 99%

See 1 more Smart Citation

Protein microarrays identify disease-specific anti-cytokine autoantibody profiles in the landscape of immunodeficiency

Rosenberg

Price

Barcenas-Morales

et al. 2016

Journal of Allergy and Clinical Immunology

Self Cite

View full text Add to dashboard Cite

Background Anti-cytokine autoantibodies (ACAAs) are pathogenic in a handful of rare immunodeficiencies. However, the prevalence and significance of other ACAAs across immunodeficiencies have not yet been described. Objective We sought to profile ACAAs in a diverse cohort of serum samples from patients with immunodeficiency and assess the sensitivity and specificity of protein microarrays for ACAA identification and discovery. Methods Highly multiplexed protein microarrays were designed and fabricated. Blinded serum samples from a cohort of 58 patients with immunodeficiency and healthy control subjects were used to probe microarrays. Unsupervised hierarchical clustering was used to identify clusters of reactivity, and after unblinding, significance analysis of microarrays was used to identify disease-specific autoantibodies. A bead-based assay was used to validate protein microarray results. Blocking activity of serum containing ACAAs was measured in vitro. Results Protein microarrays were highly sensitive and specific for the detection of ACAAs in patients with autoimmune polyendocrine syndrome type I and pulmonary alveolar proteinosis, detecting ACAA levels consistent with those in the published literature. Protein microarray results were validated by using an independent bead-based assay. To confirm the functional significance of these ACAAs, we tested and confirmed the blocking activity of select ACAAs in vitro. Conclusion Protein microarrays are a powerful tool for ACAA detection and discovery, and they hold promise as a diagnostic for the evaluation and monitoring of clinical immunodeficiency.

show abstract

Section: Discussionmentioning

confidence: 99%

mentioning

confidence: 99%

Protein microarrays identify disease-specific anti-cytokine autoantibody profiles in the landscape of immunodeficiency

Rosenberg

Price

Barcenas-Morales

et al. 2016

Journal of Allergy and Clinical Immunology

Self Cite

View full text Add to dashboard Cite

show abstract

“…Understanding and diagnosing autoimmune diseases benefit from precise knowledge of the targets of the immune system, which can serve both diagnostic and, potentially, therapeutic purposes. For these reasons, we and others have developed methods to identify targets of autoimmune disorders (1)(2)(3)(4).…”

mentioning

confidence: 99%

Systematic autoantigen analysis identifies a distinct subtype of scleroderma with coincident cancer

Shah

et al. 2016

Proc. Natl. Acad. Sci. U.S.A.

View full text Add to dashboard Cite

Scleroderma is a chronic autoimmune rheumatic disease associated with widespread tissue fibrosis and vasculopathy. Approximately two-thirds of all patients with scleroderma present with three dominant autoantibody subsets. Here, we used a pair of complementary high-throughput methods for antibody epitope discovery to examine patients with scleroderma with or without known autoantibody specificities. We identified a specificity for the minor spliceosome complex containing RNA Binding Region (RNP1, RNA recognition motif) Containing 3 (RNPC3) that is found in patients with scleroderma without known specificities and is absent in unrelated autoimmune diseases. We found strong evidence for both intra- and intermolecular epitope spreading in patients with RNA polymerase III (POLR3) and the minor spliceosome specificities. Our results demonstrate the utility of these technologies in rapidly identifying antibodies that can serve as biomarkers of disease subsets in the evolving precision medicine era.

show abstract

“…Currently, experimental techniques to identify biofluid autoantibodies are limited. The primary methods to quantify autoantibodies are radiobinding assays, immunohistochemistry, enzyme-linked immunosorbent assays (ELISA), bead-based assays and protein microarrays 10 . Most of these tools, however, require an a priori hypothesis, and are limited to single-target profiling.…”

Section: Introductionmentioning

confidence: 99%

“…Most of these tools, however, require an a priori hypothesis, and are limited to single-target profiling. High-throughput methods such as human protein microarrays have yielded novel autoantibody markers, but they are costly and require recombinant protein availability and optimization 10 . Recently, our laboratory developed an unsupervised massspectrometry-based protocol using a data-dependent acquisition approach to identify tissue-specific autoantibodies 11 .…”

Section: Introductionmentioning

confidence: 99%

Putative autoantibodies in the cerebrospinal fluid of Alzheimer’s disease patients

Lim¹,

Tsolaki²,

Batruch³

et al. 2019

F1000Res

View full text Add to dashboard Cite

Background: Recent efforts have described an immunogenic component to the pathobiology of Alzheimer’s disease (AD) and Parkinson’s disease (PD). However, current methods of studying fluid autoantibodies, such as enzyme-linked immunosorbent assays and immunohistochemistry, are hypothesis-driven and not optimal for discovering new autoantibody biomarkers by proteome-wide screening. Recently, we developed a general mass spectrometry-based approach to identify tissue-specific autoantibodies in serum, at a proteome-wide level. In this study, we adapted the method to explore novel autoantibody biomarkers in the cerebrospinal fluid (CSF) of AD and PD patients. Methods: CSF samples were obtained from 10 headache control individuals, 10 AD patients and 10 PD patients. Antibodies present in the CSF were isolated by immobilization to protein-G magnetic beads. These antibodies were incubated with a brain tissue extract, prepared from frontal cortex, pons, cerebellum and brain stem. Protein antigens captured by the protein-G magnetic bead-bound antibodies were digested with trypsin and analyzed using mass spectrometry. Autoantibody candidates were selected by 1) detection in one or less individuals of the control group and 2) identification in at least half of the patient groups. Results: There were 16 putative autoantibody biomarkers selected from the AD group. Glia-derived nexin autoantibody was detected in eight of ten AD patients and was absent in the control group. Other AD pathology-related targets were also identified, such as actin-interaction protein, quinone oxidoreductase, sushi repeat-containing protein, metalloproteinase inhibitor 2, IP3 receptor 1 and sarcoplasmic/endoplasmic reticulum calcium ATPase 2. An additional eleven autoantibody targets were also identified in the present experiment, although their link to AD is not clear. No autoantibodies in the PD group satisfied our selection criteria. Conclusion: Our unbiased mass spectrometry method was able to detect new putative CSF autoantibody biomarkers of AD. Further investigation into the involvement of humoral autoimmunity in AD and PD pathobiology may be warranted.

show abstract

Protein Microarrays: A New Tool for the Study of Autoantibodies in Immunodeficiency

Cited by 29 publications

References 77 publications

Protein microarrays identify disease-specific anti-cytokine autoantibody profiles in the landscape of immunodeficiency

Protein microarrays identify disease-specific anti-cytokine autoantibody profiles in the landscape of immunodeficiency

Systematic autoantigen analysis identifies a distinct subtype of scleroderma with coincident cancer

Putative autoantibodies in the cerebrospinal fluid of Alzheimer’s disease patients

Contact Info

Product

Resources

About