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Cpg 1826

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CpG 1826 is a synthetic oligonucleotide that contains an unmethylated CpG dinucleotide motif. It is commonly used as a tool in immunology research to stimulate the innate immune system.

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Lab products found in correlation

Synergy neo microplate reader, by Agilent Technologies (2 mentions) Ze5 cell analyzer, by Bio-Rad (2 mentions) Renca cell line, by American Type Culture Collection (1 mentions) Ivis lumina 3 imager, by PerkinElmer (1 mentions) Rat igg2a isotype control, by BioXCell (1 mentions) Jupiter c18 lc column, by Phenomenex (1 mentions) Zetasizer nano zsp, by Malvern Panalytical (1 mentions) Cd45 bv510, by BioLegend (1 mentions) Foxp3 pe cy7 fjk 16s, by Thermo Fisher Scientific (1 mentions) Ghost red 780 viability dye, by Cytek Biosciences (1 mentions) Cd11b v450 m1 70, by BD (1 mentions) Ox40 pe ox 86, by BioLegend (1 mentions) Foxp3 transcription factor staining buffer set, by Thermo Fisher Scientific (1 mentions) L ascorbic acid, by Thermo Fisher Scientific (1 mentions) Granulocyte macrophage colony stimulating factor gm csf, by GenScript (1 mentions) Penicillin streptomycin p s, by Thermo Fisher Scientific (1 mentions) Ack lysis buffer, by Thermo Fisher Scientific (1 mentions) Rpmi 1640, by Thermo Fisher Scientific (1 mentions) Mini hypodermic thermocouple probe, by Omega Engineering (1 mentions) Digital thermometer, by Omega Engineering (1 mentions)

9 protocols using cpg 1826

1

Syngeneic Renca Tumor Model with Adenoviral TRAIL

Cited 3 times
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The Renca cell line (syngeneic to BALB/c mice) was purchased from the American Type Culture Collection (ATCC; Manassas, VA, USA), engineered to express firefly luciferase, and cultured as described [4 (link),29 (link),32 (link)]. Cells were confirmed negative for mycoplasma, passaged, and used at the same passage number to limit experimental variation. Intrarenal tumor challenges and bioluminescent imaging (BLI) using an IVIS Lumina III Imager (Perkin Elmer; Waltham, MA, USA; University of Alabama at Birmingham (UAB) Small Animal Imaging Facility) were performed as described [4 (link),29 (link),32 (link)]. At day six post-tumor challenge, BLI was used to assess tumor burdens in live mice prior to therapy randomization. At day seven post-tumor challenge, mice were re-injected in the tumor-bearing kidney with either sterile saline or 109 pfu of replication-deficient adenovirus (Ad) encoding a membrane-bound version of full-length murine TRAIL protein (University of Iowa Viral Vector Core) plus 100 µg CpG1826 (Integrated DNA Technologies) (AdT/CpG treatment) and further randomized to receive no therapy, InVivoMAb polyclonal Armenian hamster IgG control (BioXCell; Lebanon, NH, USA), or InVivoMAb anti (α)-mouse CTLA-4 (clone UC10-4F10-11; BioXCell) i.p. at a dose of 100 μg/mouse on days 10, 13, and 16 post-tumor challenge.
Turbitt W.J., Boi S.K., Gibson J.T., Orlandella R.M, & Norian L.A. (2021). Diet-Induced Obesity Impairs Outcomes and Induces Multi-Factorial Deficiencies in Effector T Cell Responses Following Anti-CTLA-4 Combinatorial Immunotherapy in Renal Tumor-Bearing Mice. Cancers, 13(10), 2295.
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2

Immune-Boosting Treatment for Renal Tumors

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Intrarenal tumor challenges were performed as reported.21 (link) Renal tumor growth was confirmed at day 6 post-challenge via bioluminescent imaging (BLI). Seven days post-tumor challenge, mice were reinjected in the tumor-bearing kidney with either sterile saline or 100 µg CpG1826 (Integrated DNA Technologies) plus 109 pfu of replication-deficient adenovirus encoding a membrane-bound version of full-length murine tumor necrosis factor (TNF)-related apoptosis-inducing ligand (TRAIL) (AdTR) protein (UI Viral Vector Core). On days 10, 13 and 16 post-tumor challenge, mice received intraperitoneal injections of saline, 250 µg of Rat IgG2a Isotype Control (Clone: 2A3, BioXCell), or 250 µg of anti-PD-1 (Rat IgG2a, clone: RMP1-14, BioXCell). In some experiments, tumor-challenged mice receiving AdTR/CpG/PD-1 were injected i.p. every 3 days with saline or neutralizing antibody against CCL5 (50 µg/mouse, clone: 53405, R&D Systems) or interleukin (IL)-1β (20 mg/kg, clone: B122, BioXCell) beginning at day 5 or day −5 relative to tumor challenge, respectively.
Boi S.K., Orlandella R.M., Gibson J.T., Turbitt W.J., Wald G., Thomas L., Buchta Rosean C., Norris K.E., Bing M., Bertrand L., Gross B.P., Makkouk A., Starenki D., Farag K.I., Sorge R.E., Brown J.A., Gordetsky J., Yasin H., Garje R., Nandagopal L., Weiner G.J., Lubaroff D.M., Arend R.C., Li P., Zakharia Y., Yang E., Salem A.K., Nepple K., Marquez-Lago T.T, & Norian L.A. (2020). Obesity diminishes response to PD-1-based immunotherapies in renal cancer. Journal for Immunotherapy of Cancer, 8(2), e000725.
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3

Peptide-Functionalized Nanoparticle Vaccine

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Polyethyleneimine (PEI, branched, Mw 25000), 3‐(2‐pyridyldithio)propionic acid N‐hydroxysuccinimide ester (SPDP) were obtained from Sigma‐Aldrich. Methoxy poly(ethyleneglycol) propionic acid N‐hydroxysuccinimide (Methoxy‐PEG‐NHS, Mw 5000) was purchased from Nanocs. CpG1826 was obtained from Integrated DNA Technology. Antigen peptides used in this study were synthesized by Genemed Synthesis, which include epitopes of ovalbumin peptide SIINFEKL and CSSSIINFEKL, neo‐epitopes of MC38 colon carcinoma ASMTNMELM (Adpgk) and CSSASMTNMELM (CSS‐Adpgk), neo‐epitopes of B16F10 melanoma LCPGNKYEM (M27), VDWENVSPELNSTDQ (M30), and CSSVDWENVSPELNSTDQ (CSS‐M30). All other reagents were received from Fisher scientific unless otherwise indicated. UV–Vis absorption and fluorescence spectra were obtained using BioTek synergy neo microplate reader. GPC and HPLC were performed using Shimadzu HPLC system equipped with TSKgel G3000SWxl column (Tosoh Bioscience LLC) and Jupiter® C18 LC Column (Phenomenex), respectively. TEM images were acquired using JEOL 1400‐plus. Hydrodynamic size and zeta potential were measured using Zetasizer Nano ZSP (Malvern Panalytical). Flow cytometry was performed using ZE5 Cell Analyzer (Bio‐Rad) and the data were analyzed using FlowJo 10.5 software.
Nam J., Son S., Park K.S, & Moon J.J. (2021). Modularly Programmable Nanoparticle Vaccine Based on Polyethyleneimine for Personalized Cancer Immunotherapy. Advanced Science, 8(5), 2002577.
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4

In Vivo Immunotherapy Combination Protocol

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CpG 1826 was purchased from Integrated DNA Technologies. OX40 antibody [Anti-OX40 (CD134) antibody, rat immunoglobulin G1, OX86 clone, European Collection of Cell Cultures] was harvested and isolated from the ascites of immunodeficient mice, as previously described (33 (link)). CpG (50 µg) and/or OX40 (4 µg, 20 µg, or 100 µg) were injected IT with a 29 ½ gauge insulin syringe in 60 µL PBS every other day for three total doses (days 0, 2, 4 or days 5, 7, 9 depending on the experiment).
The following antibodies were used for flow cytometry analysis: anti-CD16/32 (93), CD45 BV510 (30-F11), CD45 FITC (30-F11), CD3 PE-Cy5 (145-2C11), CD4 BV785 (GK1.5), CD19 PE-Cy5 (6D5), CD19 APC (6D5), CD19 BV421 (6D5), Ly6G Alexa647 (1A8), IFNγ PE-Cy7 (XMG1.2), and OX40 PE (OX-86) all from BioLegend; CD8 APC-R700 (53-6.7), CD25 BB515 (PC61), NK1.1 PE-CF594 (PK136), Ly6C BV605 (AL-21), and CD11b V450 (M1/70) all from BD Biosciences; FoxP3/Transcription Factor Staining Buffer Set and FoxP3 PE-Cy7 (FJK-16s) from eBioscience. GhostRed780 Viability Dye (Tonbo Biosciences) was used for live/dead staining.
Pieper A.A., Zangl L.M., Speigelman D.V., Feils A.S., Hoefges A., Jagodinsky J.C., Felder M.A., Tsarovsky N.W., Arthur I.S., Brown R.J., Birstler J., Le T., Carlson P.M., Bates A.M., Hank J.A., Rakhmilevich A.L., Erbe A.K., Sondel P.M., Patel R.B, & Morris Z.S. (2021). Radiation Augments the Local Anti-Tumor Effect of In Situ Vaccine With CpG-Oligodeoxynucleotides and Anti-OX40 in Immunologically Cold Tumor Models. Frontiers in Immunology, 12, 763888.
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5

Photothermal Immunotherapy with Adpgk Peptide

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L-ascorbic acid was obtained from Fisher Chemical. pIC (high molecular weight, 1.5–8 kb) was purchased from InvivoGen, and CpG1826 was obtained from Integrated DNA Technology. Adpgk peptide (ASMTNMELM) was synthesized by Genemed Synthesis. RPMI 1640, penicillin-streptomycin (PS), beta-mercaptoethanol (β-ME), and ACK lysis buffer were purchased from Gibco. Fetal bovine serum (FBS) was obtained from Corning. Granulocyte-macrophage colony-stimulating factor (GM-CSF) was received from Genscript. Other chemicals were obtained from Sigma-Aldrich unless otherwise stated. UV-Vis absorption spectra were obtained using BioTek synergy neo microplate reader. Transmission electron microscope images were acquired using JEOL 1400-plus and analyzed using the ImageJ software (NIH, Bethesda, MD). Laser irradiation was performed using a 808 nm continuous wave diode laser (China Daheng Group Inc., Beijing, China). Tumor temperature was measured using a mini-hypodermic thermocouple probe coupled with digital thermometer (OMEGA engineering, Inc.). Flow cytometry was performed using ZE5 Cell Analyzer (Bio-Rad) and the data were analyzed using FlowJo 10.5 software.
Nam J., Son S., Park K.S, & Moon J.J. (2021). Photothermal therapy combined with neoantigen cancer vaccination for effective immunotherapy against large established tumors and distant metastasis. Advanced therapeutics, 4(8), 2100093.
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6

Bone Marrow Dendritic Cell Generation

Cited 1 time
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Bone marrow-derived DC (DC) were generated as previously described (45 (link)), with minor changes. In brief, bone marrow was flushed from femurs and tibia from either young BALB/c ByJ or from Il6 −/− BALB/c ByJ mice and plated at 8 × 105 cells/ml in complete RPMI 1640 plus 10 ng/ml recombinant murine GM-CSF (PeproTech, Rocky Hill, NJ). On days 3 and 5, half of the culture media was replaced with fresh complete RPMI 1640 + GM-CSF. On day 6 the TLR9 agonist CpG 1826 (1μM; type B unmethylated CpG-oligodeoxynucleotides [ODNs]) (Integrated DNA Technologies, Coralville, IA), was added to the “activated” DC group followed 3 hours later by Poly I:C (10 μg/ml; Invivogen, San Diego, CA), a combination shown to have a synergistic adjuvant effect (31 (link)). On day 7 non-adherent unactivated DC and TLR agonist activated *DC were harvested and enriched by positive selection using directly conjugated CD11c magnetic beads and a MACS column, as per manufacturer’s instructions (Miltenyi Biotec).
Brahmakshatriya V., Kuang Y., Devarajan P., Xia J., Zhang W., Vong A.M, & Swain S.L. (2017). IL-6 production by TLR-activated APC broadly enhances aged cognate CD4 helper and B cell antibody responses in vivo. Journal of immunology (Baltimore, Md. : 1950), 198(7), 2819-2833.
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7

Conjugated Polymer-based Nanovaccine Formulation

Cited 1 time
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Conjugated polymer between PEI and PEG (PEI-PEG) was prepared using branched PEI (Sigma-Aldrich, molecular weight ~25,000) and methoxy poly(ethyleneglycol) propionic acid N-hydroxysuccinimide (Nanocs, molecular weight 5000) as reported previously [27 (link)]. Antigen peptides employed in this study include SIINFEKL, CSSSIINFEKL, ASMTNMELM (Adpgk), CSSASMTNMELM (CSS-Adpgk), and LCPGNKYEM (M27). All antigen peptides were obtained from Genemed Synthesis. For PEI conjugates (PEI-PEG/CSS-antigen or PEI-PEG/M27), PEI-PEG was dissolved in DMSO and added with 3-(2-Pyridyldithio)propionic acid N-hydroxysuccinimide ester (SPDP, Sigma-Aldrich) to create thiol-reactive disulfide bond. After stirring for 3h, the mixture was reacted with CSS-antigen overnight, followed by dialysis-purification using Amicon ultra 10 kDa MW cutoff centrifugal filters. The resulting PEI conjugate was vigorously mixed with CpG (CpG 1826, Integrated DNA Technology) in PBS at varying PEI conjugate/CpG weight ratio of 0.5, 1, 2, or 3 to form respective NP Vacc by CpG nano-complexation.
Park K.S., Nam J., Son S, & Moon J.J. (2021). Personalized combination nano-immunotherapy for robust induction and tumor infiltration of CD8+ T cells.. Biomaterials, 274, 120844.
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8

Nanoparticle-based Vaccine Formulation

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Poly(lactic
coglycolic acid) 50:50 (0.45–0.60
dL/g, MW = 38,000–54 000
Da, ester-terminated) and poly(vinyl alcohol) (87–89% hydrolyzed MW = 13,000–23,000) were purchased from
Milipore-Sigma. CpG 1826 was created with a thioester backbone and
purchased from Integrated DNA Technologies. Purified ovalbumin was
purchased from Worthington Biomedical. Honokiol was obtained from
TCI Chemicals (Japan), and capsaicin was purchased from Millipore
Sigma. SN50 was created using a Liberty Blue peptide synthesizer and
purified via HPLC. All other materials and assays were purchased from
commercial sources.
Cassaidy B.J., Moser B.A., Solanki A., Chen Q., Shen J., Gotsis K., Lockhart Z., Rutledge N., Rosenberger M.G., Dong Y., Davis D, & Esser- Kahn A.P. (2024). Immune Potentiation of PLGA Controlled-Release Vaccines for Improved Immunological Outcomes. ACS Omega, 9(10), 11608-11614.
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9

Nanotoxoid Preparation and Vaccination

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α7-Neuro-2a cell membrane-coated nanoparticles (α7-NPs) were incubated with αBgt (Millipore Sigma) or Alexa Fluor 647-labeled αBgt at the prescribed polymer to αBgt weight ratios at room temperature for 2 h to prepare nanotoxoids (α7-NTs). For in vivo studies, α7-NTs were adjusted to a polymer concentration of 5 mg/mL in 10 % sucrose in water. For vaccination studies, α7-NTs were mixed with CpG 1826 (Integrated DNA Technologies) at a 20:1 polymer to DNA weight ratio right before administration. To generate a heat-inactivated αBgt (iαBgt) control, the toxin was heated at 99 °C for 12 h.
Guo Z., Zhu A.T., Wei X., Jiang Y., Yu Y., Noh I., Gao W., Fang R.H, & Zhang L. (2024). A genetically engineered neuronal membrane-based nanotoxoid elicits protective immunity against neurotoxins. Bioactive Materials, 38, 321-330.
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