ODN 1018 VacciGrade™

CpG ODN, type B (human/mouse) - TLR9-based vaccine adjuvant

TLR9 activation with ODN 1018

CpG ODN 1018, also known as CpG 1018® or 1018 ISS,  is a synthetic immunostimulatory oligonucleotide (ODN) that contains unmethylated CpG dinucleotides. It is currently the only FDA-approved CpG-based adjuvant for human vaccine use. 

ODN 1018 is a class B CpG ODN containing CpG motifs that are able to bind and activate both the rodent and human Toll-like receptor 9 (TLR9) [1]. It has been extensively tested in the clinic for its efficacious immunological effects, such as the production of inflammatory cytokines in macrophages and natural killer (NK) cells, the promotion of Th1 cell responses, and its involvement in B cell proliferation [2].

 More details

Interestingly, ODN 1018 VacciGrade™ successfully activates the murine (m)TLR9, but not the human (h)TLR9 in HEK-derived cells [in-house data]. In monocytic THP1-Dual™ hTLR9 cells, however, ODN 1018 VacciGrade™ shows a strong hTLR9-mediated NF-κB and IRF induction (see figures). ODN 1018 VacciGrade™ is a high-quality pre-clinical grade, suitable for in vivo use.

ODN 1018  VacciGrade™​ is for research use only, and not for human or veterinary use. It is not a pharmaceutical preparation fit for vaccine manufacturing.
CpG 1018® is a registered trademark of Dynavax Technologies and is used for comparative purposes only. ODN 1018 is not made by, affiliated with, sponsored by, or endorsed by Dynavax Technologies. Dynavax Technologies has not evaluated or approved the representations contained herein.

References:

1. Campbell JD, 2017. Development of the CpG Adjuvant 1018: A Case Study. Methods Mol Biol. 1494:15-27.
2. Eng NF, et al., 2013. The potential of 1018 ISS adjuvant in hepatitis B vaccines: HEPLISAV™ review. Hum Vaccin Immunother. (8):1661-72.

Figures

ODN 1018 VacciGrade™ is a potent activator of murine TLR9 in HEK-derived cells. HEK-Blue™ murine (m)TLR9 and HEK-Blue™ human (h)TLR9 cells were incubated in HEK-Blue™ Detection reagent with increasing concentrations of ODN 1018 VacciGrade™. After 24 hours, SEAP activity in the cell culture supernatant was assessed. Data are shown as optical density (OD) at 650 nm (mean ± SEM).

ODN 1018 VacciGrade™ is a potent activator of human TLR9 in THP-1-derived cells. THP1-Dual™ and THP1-Dual™ human (h)TLR9 cells were incubated with increasing concentrations of ODN 1018 VacciGrade™. After 24h, the hTLR9-induced (A) NF-κB and (B) IRF responses were assessed by measuring SEAP and Lucia activity using QUANTI-Blue™ and QUANTI-Luc™, respectively. Data are shown as optical density (OD) at 650 nm or in fold response over non-induced cells (mean + SEM).

Specifications

ODN 1018 sequence: 5′-TGACTGTGAACGTTCGAGATGA-3′ (22-mer)

Description: TLR9 agonist VacciGrade™

Synonyms: CpG 1018®, 1018 ISS

CAS number: 937402-51-2

Polarization of adaptive immune response: Th1 response

Solubility: 2 mg/ml in physiological water

Working concentration: 1 ng/ml - 10 µg/ml

Quality control:

• Sterility guaranteed
• The absence of bacterial contamination (lipoproteins & endotoxins) has been confirmed using HEK-Blue™ TLR2 and HEK-Blue™ TLR4 cells
• Endotoxin level < 1 EU/mg (measurement by kinetic chromogenic LAL assay)

Contents

• 1 mg (139.8 nmol) of sterile lyophilized ODN 1018 VacciGrade™.
• 10 ml sterile endotoxin-free physiological water (NaCl 0.9%).

ODN 1018 VacciGrade™ is shipped at room temperature.

ODN 1018 VacciGrade™ should be stored at -20°C.

Product is stable for 6 months at -20°C when properly stored.

Avoid repeated freeze-thaw cycles.

VacciGrade™

VacciGrade™ is a high-quality pre-clinical grade. 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. The level of bacterial contaminants (endotoxins and lipoproteins) in each lot is verified using a LAL assay and/or a TLR2 and TLR4 reporter assay.
*Except for LPS VacciGrade™, which is prepared in a laminar flow hood dedicated to LPS.

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]. 

CpG ODN 1018

ODN 1018, also known as 1018 immunostimulatory sequence (ISS), is a type B CpG ODN with the sequence 5′-TGACTGTGAACGTTCGAGATGA-3′. It mimics bacterial single-stranded (ss) DNA to activate innate immunity through binding to TLR9. The first CpG motif (underlined) is a sequence active on mouse TLR9, whereas the second CpG motif (bold) is active on human and non-human primate TLR9 [6]. Type B CpG ODNs contain a full phosphorothioate backbone with one or more CpG dinucleotides and were described to strongly activate the Th1 cell and B cell populations [4,6]. 

ODN 1018 has been extensively tested in the clinic for its efficacious immunological effects, such as the production of IL-12, IL-18, and IFN-γ from macrophages and natural killer (NK) cells, promoting Th1 responses, and its involvement in antibody production and B cell proliferation [7]. The GMP-produced CpG 1018® is currently the only FDA-approved CpG adjuvant for human vaccine use. It is a component of several approved vaccines, including the licensed hepatitis B virus vaccine HEPLISAV-B® [8]. Various pre-clinical and clinical trials are ongoing using ODN 1018 alone or together with alum in vaccine strategies against infectious diseases (e.g. SARS-CoV-2) or cancer [8]. Since TLR9 is an important regulator of microglia, ODNs have been extensively tested for their potential to stimulate the clearance of amyloid pathology and treat Alzheimer’s disease (AD) [9]. Clinical trials are ongoing investigating the impact of CpG 1018® alone in early AD patients (NCT05606341) [10].

In summary, the mechanism of ODN 1018 and how it exerts its adjuvant effect remains to be elucidated. Advances of the past decades in understanding innate immunity have brought a wider interest in understanding how ODN 1018 works and how it might be improved [7].

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. Campbell JD, 2017. Development of the CpG Adjuvant 1018: A Case Study. Methods Mol Biol. 1494:15-27.
7. Eng NF, et al., 2013. The potential of 1018 ISS adjuvant in hepatitis B vaccines: HEPLISAV™ review. Hum Vaccin Immunother.(8):1661-72.
8. Strohmeier S, et al., 2022. A CpG 1018 adjuvanted neuraminidase vaccine provides robust protection from influenza virus challenge in mice. NPJ Vaccines. 2022 Jul 22;7(1):81.
9. Patel AG, et al., 2021.  Innate immunity stimulation via CpG oligodeoxynucleotides ameliorates Alzheimer's disease pathology in aged squirrel monkeys. Brain. 2021 Aug 17;144(7):2146-2165. 
10. clinicaltrials.gov