Cpg oligodeoxynucleotide

Manufactured by InvivoGen

Sourced in United States, France

CpG oligodeoxynucleotides are short synthetic DNA sequences that contain unmethylated CpG motifs. They can be used as molecular tools to stimulate the immune system.

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Lab products found in correlation

Lps from escherichia coli, by Merck Group (2 mentions) Ionomycin, by Merck Group (2 mentions) Poly 1 c, by InvivoGen (2 mentions) Facsaria 2, by BD (2 mentions) Pam3csk4, by InvivoGen (2 mentions) Phorbol myristate acetate (pma), by Cell Signaling Technology (1 mentions) Carboxyfluorescein succinimidyl ester (cfse), by BioLegend (1 mentions) Cd19 pe cy7 clone hib19, by BioLegend (1 mentions) F ab 2 anti igm antibody, by Jackson ImmunoResearch (1 mentions) Zombie aqua amcyan viability dye, by BioLegend (1 mentions) Cd20 percp cy5.5 clone 2h7, by BioLegend (1 mentions) Carboxyfluorescein diacetate succinimidyl ester cfse, by BioLegend (1 mentions) D desthiobiotin, by Merck Group (1 mentions) Z leu leu leu al, by Merck Group (1 mentions) Puromycin dihydrochloride, by GoldBio (1 mentions) Pe conjugated anti mouse igg antibody, by BD (1 mentions) Brefeldin a, by Merck Group (1 mentions) Phorbol myristate acetate (pma), by Merck Group (1 mentions) Gm csf, by Thermo Fisher Scientific (1 mentions)

Close spelling variants of this product

CpG-A oligodeoxynucleotide 1585 (2 citations) CpG oligodeoxynucleotide (ODN) 2006 (3 citations) CpG oligodeoxynucleotides (ODN 2006, (3 citations) CpG-oligodeoxynucleotide (CpG-ODN) (2 citations) Murine CpG oligodeoxynucleotide (CpG) TLR9 ligand (ODN 1585; Class A) (3 citations) CpG oligodeoxynucleotide (ODN) 1826 VacciGrade (2 citations) CpG- oligodeoxynucleotides (ODN) (4 citations) CpG oligodeoxynucleotide 2006 (6 citations) Class B CpG oligodeoxynucleotides (2 citations)

8 protocols using cpg oligodeoxynucleotide

Stimulation of B and T Cells

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We stimulated 1.5 × 10⁵ to 2.0 × 10⁵ B cells in 96-well plates in supplemented RPMI-1640 medium with antibodies or immunostimulants during the indicated times. Stimuli included CpG oligodeoxynucleotides (ODN 2006) for B cells (InvivoGen), LPS from Escherichia coli (Sigma-Aldrich), anti-human F(ab’)2 IgM and F(ab’)2 IgG (Southern Biotech), ionomycin (EMD Millipore), PMA (Cell Signaling Technology), and cross-linked human CD40L (Miltenyi). After stimulation, cells were washed and resuspended in PBS before flow cytometry analysis. For antigen-specific stimulation, 100 antigen-coated beads per B cell were added in the supernatant.
For Jurkat T cells stimulation, 2 × 10⁶ cells were plated in 96-well plates and stimulated with 1 μg/mL CD3/CD8 antibodies (Invitrogen) or with TransAct (Myltenyi Biotech) according to the manufacturer’s instructions. For kinetics assessments, to remove stimulant molecules, plates were washed three times with 200 μL PBS.

Page A., Delles M., Nègre D., Costa C., Fusil F, & Cosset F.L. (2023). Engineering B cells with customized therapeutic responses using a synthetic circuit. Molecular Therapy. Nucleic Acids, 33, 1-14.

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B cell proliferation upon DENV-2 infection

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B cells isolated from healthy donors were labeled with carboxyfluorescein diacetate succinimidyl ester (CFSE) (Biolegend) and either infected with DENV-2 at an MOI of 5 or remained uninfected. After 2 h of inoculation, cells were washed to remove the inoculum and stimulated with CpG oligodeoxynucleotides (1 μg/ml; Invivogen, San Diego, CA, USA) and F(ab′)₂ anti-IgM antibody (4 μg/ml; Jackson ImmunoResearch, PA, USA) or remained unstimulated. Cells were cultured in RPMI supplemented with 10% FBS for 6 days. The cells were then harvested and stained with Zombie Aqua (AmCyan) viability dye (BioLegend) followed by CD19 PE/Cy7 (clone HIB19) and CD20 PerCp/Cy5.5 (clone 2H7) to identify live B cells and analyzed for the expression of CFSE in the FITC channel. Proliferation was measured as the percentage of B cells with decreased intensity of CFSE compared to unstimulated B cells.

Upasani V., Vo H.T., Auerswald H., Laurent D., Heng S., Duong V., Rodenhuis-Zybert I.A., Dussart P, & Cantaert T. (2021). Direct Infection of B Cells by Dengue Virus Modulates B Cell Responses in a Cambodian Pediatric Cohort. Frontiers in Immunology, 11, 594813.

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Reagents for Immune Stimulation

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Escherichia coli 0111:B4 lipopolysaccharide (LPS), zymosan from Saccharomyces cerevisiae, Z-Leu-Leu-Leu-al (MG-132), and D-desthiobiotin were purchased from Sigma-Aldrich (St. Louis, MO, USA). CpG oligodeoxynucleotides were purchased from Invivogen (Toulouse, France). N-Acetyl-Leu-Leu-Norleu-al (ALLN) was purchased from Santa Cruz Biotechnology (Dallas, TX, USA). Puromycin dihydrochloride was purchased from Gold Biotechnology (St. Louis, MO, USA).

Aslanidis A., Karlstetter M., Scholz R., Fauser S., Neumann H., Fried C., Pietsch M, & Langmann T. (2015). Activated microglia/macrophage whey acidic protein (AMWAP) inhibits NFκB signaling and induces a neuroprotective phenotype in microglia. Journal of Neuroinflammation, 12, 77.

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Antigen cross-presentation in A549 cells

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A549 cells (antigen donor cells) were mock-infected or infected with IAV virus or IAV-OVA virus (MOI = 10) for 10 h prior to γ-irradiation (50 Gy) to inhibit the proliferation of viruses and induce apoptosis of the cells. After washing, the irradiated cells were incubated with Mutu cells (GFP⁺) at a ratio of 1:1 or 1:2 in the presence of 500 nM CpG oligodeoxynucleotides (InvivoGen) at 37 °C and harvested at the designated time. For preliminary analysis of cross-presentation, the Mutu cells were then co-cultured with VPD450-labeled OT-1 CD8⁺ T cells at ratio of 1:1 at 37 °C for 24 h. OT-1 T-cell proliferation was assessed based on the VPD450 dye dilution in CD8⁺ T cells. Alternatively, the Mutu cells were stained with monoclonal 25-D1.16 antibody, specific for the H-2K^b bound peptide OVA_257–264 (SIINFEKL), followed by a PE-conjugated anti-mouse IgG antibody (BD Biosciences), and analyzed by flow cytometry.

Wu T., Guan J., Handel A., Tscharke D.C., Sidney J., Sette A., Wakim L.M., Sng X.Y., Thomas P.G., Croft N.P., Purcell A.W, & La Gruta N.L. (2019). Quantification of epitope abundance reveals the effect of direct and cross-presentation on influenza CTL responses. Nature Communications, 10, 2846.

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PBMC Stimulation Assay

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2 x 10⁶ overnight rested PBMC were seeded in polypropylene U-bottom 96-well microtiter plates (Fisher Scientific, Hampton, USA) with CTL-Test™ Medium and either stimulated with CpG oligodeoxynucleotides (10 µg/mL) (InvivoGen, Toulouse, France), PMA (Phorbol-12-myristat-13-acetat) (30 ng/mL) (Sigma-Aldrich, St. Louis, USA), and Ionomycin (1 µg/mL) (Sigma-Aldrich, St. Louis, USA) (referred to as “CpG+P/I”) or left unstimulated as a medium control to define background activity. After 1 h of incubation at 37°C in 5% CO₂, 10 µg/mL of secretion blocker Brefeldin A (Sigma-Aldrich, St. Louis, USA) in a total volume of 50 µL CTL-Test™ medium was added to each well. Stimulation was stopped after 4 h by transferring the plates to 4°C overnight.

Körber N., Pohl L., Weinberger B., Grubeck-Loebenstein B., Wawer A., Knolle P.A., Roggendorf H., Protzer U, & Bauer T. (2021). Hepatitis B Vaccine Non-Responders Show Higher Frequencies of CD24highCD38high Regulatory B Cells and Lower Levels of IL-10 Expression Compared to Responders. Frontiers in Immunology, 12, 713351.

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Immune Mediators and Stimulants

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Recombinant human CXCL8(6-77), TNF-α, interferon-γ (IFN-γ), IL-1β and granulocyte-macrophage colony-stimulating factor (GM-CSF) were purchased from Peprotech (Rocky Hill, NJ, USA). The bacterial tripeptide N-formyl-methionyl-leucyl-phenylalanine (fMLF), peptidoglycan from Staphylococcus aureus (PGN), phorbol 12-myristate 13-acetate (PMA), lipopolysaccharide (LPS) from Klebsiella pneumoniae and LPS from Escherichia coli were purchased from Sigma-Aldrich (St. Louis, MO, USA). CpG oligodeoxynucleotides were purchased from InvivoGen (San Diego, CA, USA).

Blanter M., Cambier S., De Bondt M., Vanbrabant L., Pörtner N., Abouelasrar Salama S., Metzemaekers M., Marques P.E., Struyf S., Proost P, & Gouwy M. (2022). Method Matters: Effect of Purification Technology on Neutrophil Phenotype and Function. Frontiers in Immunology, 13, 820058.

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Isolation and Characterization of Murine Bone Marrow and Peritoneal Neutrophils

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Mature murine bone marrow PMNs were isolated from mouse femurs and tibias by discontinuous Percoll gradient centrifugation as previously reported.^{19 (link)} This preparation was then sorted by a FACSAriaII (Becton Dickinson, Franklin Lakes, NJ) based on live (7AAD^neg), Ly6G⁺, and F4/80^{6 (link)} to yield a population that was >95% pure with 0% F4/80⁺ cells (Figure 7A). Peritoneal PMNs were harvested by peritoneal lavage 4h after i.p. Brewer's thioglycollate (2 ml, 4%). Peritoneal exudate macrophages were harvested by peritoneal lavage 96h after i.p. Brewer's thioglycollate and then plated for 24h before stimulation. Macrophages were stimulated with media, Pam3CSK4 (Invivogen, San Diego, CA; 100ng/ml), Poly I:C (Invivogen, 10μg/ml), LPS (List Biologicals, Campbell, CA; 10ng/ml), or CpG oligodeoxynucleotide (Invivogen; 10Qg /ml) for 24h and cell supernatants harvested for cytokine analysis.

Gowdy K.M., Madenspacher J.H., Azzam K.M., Gabor K.A., Janardhan K.S., Aloor J.J, & Fessler M.B. (2014). Key Role for Scavenger Receptor B-I in the Integrative Physiology of Host Defense during Bacterial Pneumonia. Mucosal immunology, 8(3), 559-571.

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Isolation and Characterization of Murine Bone Marrow and Peritoneal Neutrophils

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