A Short DNA Aptamer That Recognizes TNFα and Blocks Its Activity <i>in Vitro</i>

Orava, Erik W.; Jarvik, Nick; Shek, Yuen Lai; Sidhu, Sachdev S.; Gariépy, Jean

doi:10.1021/cb3003557

Cited by 84 publications

(62 citation statements)

References 51 publications

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“…The vast majority of aptamer applications have involved using the aptamer as a substitute for an antibody in an immunoassay-type platform although a few interesting exceptions exist using electrochemical-based detection platforms and others including aptamer-labeled pores [138, 139]. Reported aptamers that bind cytokines include: interferon-γ (IFN-γ) [140]; interleukin-6 (IL-6) [141]; IL-17 [142]; IL-32 [143]; oncostatin M, a member of the IL-6 family [144]; platelet derived growth factor (PDGF) [145]; tumor necrosis factor-α [146, 147], and vascular endothelial growth factor (VEGF) [148]. Among these cytokines, new analytical chemistry assays or devices have been described using aptamers against IFN-γ, PDGF and VEGF.…”

Section: Aptamer Assaysmentioning

confidence: 99%

Bioanalytical chemistry of cytokines – A review

Stenken

Poschenrieder

2015

Analytica Chimica Acta

237

172

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Cytokines are bioactive proteins produced by many different cells of the immune system. Due to their role in different inflammatory disease states and maintaining homeostasis, there is enormous clinical interest in the quantitation of cytokines. The typical standard methods for quantitation of cytokines are immunoassay-based techniques including enzyme-linked immusorbent assays (ELISA) and bead-based immunoassays read by either standard or modified flow cytometers. A review of recent developments in analytical methods for measurements of cytokine proteins is provided. This review briefly covers cytokine biology and the analysis challenges associated with measurement of these biomarker proteins for understanding both health and disease. New techniques applied to immunoassay-based assays are presented along with the uses of aptamers, electrochemistry, mass spectrometry, optical resonator-based methods. Methods used for elucidating the release of cytokines from single cells as well as in vivo collection methods are described.

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Section: Aptamer Assaysmentioning

confidence: 99%

Bioanalytical chemistry of cytokines – A review

Stenken

Poschenrieder

2015

Analytica Chimica Acta

237

172

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“…We have recently shown that functional DNA aptamers targeting TNF-α 42 and CEA, 43 do not trigger cytokine responses linked to innate immunity. Consistently, the PEGylated aptamers used in this study did not activate a TLR9 innate immune response.…”

Section: Discussionmentioning

confidence: 99%

Agonistic CD200R1 DNA Aptamers Are Potent Immunosuppressants That Prolong Allogeneic Skin Graft Survival

Prodeus

Cydzik

Abdul-Wahid

et al. 2014

Molecular Therapy - Nucleic Acids

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CD200R1 expressed on the surface of myeloid and lymphoid cells delivers immune inhibitory signals to modulate inflammation when engaged with its ligand CD200. Signalling through CD200/CD200R1 has been implicated in a number of immune-related diseases including allergy, infection, cancer and transplantation, as well as several autoimmune disorders including arthritis, systemic lupus erythematosus, and multiple sclerosis. We report the development and characterization of DNA aptamers, which bind to murine CD200R1 and act as potent signalling molecules in the absence of exogenous CD200. These agonistic aptamers suppress cytotoxic T-lymphocyte induction in 5-day allogeneic mixed leukocyte culture and induce rapid phosphorylation of the CD200R1 cytoplasmic tail thereby initiating immune inhibitory signalling. PEGylated conjugates of these aptamers show significant in vivo immunosuppression and enhance survival of allogeneic skin grafts as effectively as soluble CD200Fc. As DNA aptamers exhibit inherent advantages over conventional protein-based therapeutics including low immunogenicity, ease of synthesis, low cost, and long shelf life, such CD200R1 agonistic aptamers may emerge as useful and safe nonsteroidal anti-inflammatory therapeutic agents.

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“…Target-specific, short, single-stranded DNA (ssDNA) molecules, called aptamers, are auspicious ligands for numerous in vivo applications. [1][2][3][4] Using the combinatorial chemistry process SELEX (Systematic Evolution of Ligands by EXponential enrichment), aptamers can be selected from a large combinatorial pool of sequences, called start library, against various target molecules such as toxins, [5] antibiotics, [6] amino acids, [7] peptides, [8] proteins, [9] or whole living cells. [10][11][12] During the in vivo application, synthetic oligonucleotides come in contact with the cells of the patient's immune system.…”

Section: Introductionmentioning

confidence: 99%

In vitro test system for evaluation of immune activation potential of new single‐stranded DNA‐based therapeutics

Avci‐Adali

Hann

Michel

et al. 2014

Drug Testing and Analysis

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Aptamers are synthetic single-stranded DNA (ssDNA) molecules with the ability to fold into complex three-dimensional structures. They can bind their targets with a high selectivity and affinity, thus they have an enormous potential as therapeutic agents. However, since aptamers are synthetic and especially since certain sequences can increasingly bind to the pattern recognition receptors of the immune cells when applied in vivo, they can induce an immune activation. Here, we established a real-time quantitative reverse transcription polymerase chain reaction (qRT-PCR) based assay to evaluate aptamers-induced immune activation prior to in vivo studies. Human whole blood or plasmacytoid dendritic cell line (PMDC05) were incubated with CpG, R10-60 aptamer, start library, or a CpG containing aptamer. After 2 and 4 h, cytokine expression was measured using qRT-PCR to determine immune reaction against different aptamers. CpG containing a phosphorothioate backbone led to a significant up-regulation of CCL-7, IFN-1α, IFN-1β in whole blood after 4 h. Compared to the samples without ssDNA, significantly higher TNF-α expression was detected after the R10-60 aptamer incubation for 4 h. The stimulation of PMDC05 cells with different ssDNA enabled more sensitive detection of aptamer sequence specific immune activation. After 4 h, CpG led to a significantly higher expression of CCL-8, CXCL-10, IL-1β, IL-6, IL-8, IFN-1β, and TNF-α. R10-60 aptamer caused a significant up-regulation of IL-1β, IFN-1β, and TNF-α. Negative control aptamers did not induce an immune activation. The use of this assay before starting with in vivo studies will facilitate the in vitro prediction of immune activation potential of aptamers.

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A Short DNA Aptamer That Recognizes TNFα and Blocks Its Activity in Vitro

Cited by 84 publications

References 51 publications

Bioanalytical chemistry of cytokines – A review

Bioanalytical chemistry of cytokines – A review

Agonistic CD200R1 DNA Aptamers Are Potent Immunosuppressants That Prolong Allogeneic Skin Graft Survival

In vitro test system for evaluation of immune activation potential of new single‐stranded DNA‐based therapeutics

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