ODN 1585 VacciGrade™
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Cat.code:
vac-1585-1
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ABOUT
CpG ODN, type A (mouse) - TLR9-based vaccine adjuvant
ODN 1585 VacciGrade™ is a Class A CpG oligonucleotide (ODN), activator of mouse Toll-like receptor 9 (TLR9) [1]. ODN 1585 is characterized by a central, phosphodiester, CpG-containing palindromic motif and phosphorothioate-modified poly-G sequences at the 3′ and 5′ ends. CpG motifs mimic microbial DNA and act as immunostimulants via TLR9 [2].
ODN 1585 has been optimized to indirectly activate natural killer (NK) cells while it has little effect on cytokine secretion or B cell proliferation [1]. ODN 1585 successfully induces regression of established melanomas in mice due to NK lytic activity [1].
ODN 1585 activates mouse TLR9 in HEK-Blue™ mTLR9 reporter cells, . Interestingly, ODN 1585 can activate the human (h)TLR9-mediated NF-κB and IRF pathways in THP1-Dual™ hTLR9 reporter cell line. This monocytic cell line overexpresses the human TLR9 gene and features two reporter genes, an NF-κB-inducible SEAP and an IRF-inducible Lucia luciferase (see figures).
ODN 1585 VacciGrade™ is a high-quality pre-clinical grade, suitable for in vivo use.
A standard grade ODN 1585 for in vitro assays is also available.
Get more information about CpG ODNs Classes.
1. Ballas ZK. et al., 2001. Divergent therapeutic and immunologic effects of oligodeoxynucleotides with distinct CpG motifs. J Immunol. 167(9):4878-86.
2. Kumagai Y. et al., 2008. TLR9 as a key receptor of the recognition of DNA. Adv. Drug. Deliv. Rev. 60(7):795-804.
All products are for internal research use only, and not for human or veterinary use.
VacciGrade™
VacciGrade™ is a high-quality pre-clinical grade, suitable for in vivo studies. VacciGrade™ products are filter-sterilized (0.2 µm) and filled under strict aseptic conditions in a clean room. The absence of bacterial contamination is assessed by a sterility test using a pharmacopeia-derived assay.
SPECIFICATIONS
Specifications
TLR9
5’- ggGGTCAACGTTGAgggggg -3’ (20 mer)
Note: Regular and capital letters represent phosphorothioate and phosphodiester bonds, respectively, bold letters represent CpG dinucleotides.
20 - 50 μg/mouse
2 mg/ml in physiological water
Sterility guaranteed
<1 EU/ml (measurement by kinetic chromogenic LAL assay)
Th1 response, TLR9 activation in cellular assays (tested)
Each lot is functionally tested and validated using cellular assays.
CONTENTS
Contents
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Product:ODN 1585 VacciGrade™
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Cat code:vac-1585-1
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Quantity:1 mg
10 ml sterile endotoxin-free physiological water (NaCl 0.9%)
Shipping & Storage
- Shipping method: Room temperature
- -20°C
- Avoid repeated freeze-thaw cycles
Storage:
Caution:
Details
CpG ODNs
Synthetic oligodeoxynucleotides containing unmethylated CpG motifs (CpG ODNs), such as ODN 1018, have been extensively studied as adjuvants [1]. These CpG motifs are present at a 20-fold greater frequency in bacterial DNA compared to mammalian DNA [2]. CpG ODNs are recognized by the Toll-like receptor 9 (TLR9), which is expressed on human B cells and plasmacytoid dendritic cells (pDCs), thereby inducing Th1-dominated immune responses [3]. Pre-clinical studies, conducted in rodents and non-human primates, as well as human clinical trials, have demonstrated that CpG ODNs can significantly improve vaccine-specific antibody responses [1]. Three types of stimulatory CpG ODNs have been identified, types A, B, and C, which differ in their immune-stimulatory activities [4-5].
Toll-like receptor 9
The Toll-like Receptor 9 (TLR9) is an endosomal receptor that triggers NF-κB- and interferon regulatory factor (IRF)-mediated pro-inflammatory responses upon the recognition of unmethylated cytosine-phosphorothioate-guanosine (CpG) forms of DNA [6-8]. Unmethylated CpG dinucleotides are a hallmark of microbial (bacterial, viral, fungal, and parasite) DNA, as well as mitochondrial self-DNA [8,9]. These TLR9 agonists can be mimicked by synthetic oligonucleotides containing CpG motifs (CpG ODNs), which have been extensively studied to improve adaptive immune responses in the context of vaccination [6,8].
TLR9 is mainly expressed in subsets of Dendritic Cells and B cells of all mammals. In rodents, but not in humans, TLR9 is also expressed in monocytes and macrophages [8]. The structure of the receptor varies by 24% between human TLR9 (hTLR9) and mouse TLR9 (mTLR9) [8]. They recognize different CpG motifs, the optimal sequences being GTCGTT and GACGTT for hTLR9 and mTLR9, respectively [10].
References:
1. Steinhagen F. et al., 2011. TLR-based immune adjuvants. Vaccine 29(17):3341-55.
2. Hemmi H. et al., 2000. A Toll-like receptor recognizes bacterial DNA. Nature 408:740-5.
3. Coffman RL. et al., 2010. Vaccine adjuvants: Putting innate immunity to work. Immunity 33(4):492-503.
4. Krug A. et al., 2001. Identification of CpG oligonucleotide sequences with high induction of IFN-alpha/beta in plasmacytoid dendritic cells. Eur J Immunol, 31(7): 2154-63.
5. Marshall JD. et al., 2005. Superior activity of the type C class of ISS in vitro and in vivo across multiple species. DNA Cell Biol. 24(2):63-72.
6. Kumagai Y. et al., 2008. TLR9 as a key receptor of the recognition of DNA. Adv. Drug. Deliv. Rev. 60(7):795-804.
7. Heinz L.X. et al., 2021. TASL is the SLC15A4-associated adaptor for IRF5 activation by TLR7-9. Nature. 581(7808):316-322.
8. Kayraklioglu N. et al., 2021. CpG oligonucleotides as vaccine adjuvants. DNA Vaccines: Methods and Protocols. Methods in Molecular Biology. Vol. 2197. p51-77.
9. Kumar V., 2021. The trinity of cGAS, TLR9, and ALRs: guardians of the cellular galaxy against host-derived self-DNA. Front. Immunol. 11:624597.
10. Bauer S. et al., 2001. Human TLR9 confers responsiveness to bacterial DNA via species-specific CpG motif recognition. Proc Natl Acad Sci USA, 98(16):9237-42.
DOCUMENTS
Documents
Technical Data Sheet
Validation Data Sheet
Safety Data Sheet
Certificate of analysis
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