Linear doggybone DNA vaccine induces similar immunological responses to conventional plasmid DNA independently of immune recognition by TLR9 in a pre-clinical model

Allen, Alexandra J; Wang, Chuan; Caproni, Lisa J.; Sugiyarto, Gessa; Harden, Elena L.; Douglas, Leon; Duriez, Patrick J.; Karbowniczek, Kinga; Extance, Jon; Rothwell, Paul J.; Orefo, Ifeayinwa; Tite, J P; Stevenson, Freda K.; Ottensmeier, Christian; Savelyeva, Natalia

doi:10.1007/s00262-017-2111-y

Cited by 31 publications

(27 citation statements)

References 45 publications

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“…P.Dom without the epitope served as a control. One injection containing 50 mg of DNA in PBS was given and any repeat doses were given 3 weeks after initial immunization (27). NOX4 inhibitor GKT137831 (Setanaxib, Genkyotex SA) was reconstituted in 1.2% methyl cellulose (Sigma) with 0.1% Polysorbate (Sigma) and administered to mice by oral gavage 5Â/week at 40 mg/kg when tumors were palpable.…”

Section: Murine Tumor Modelsmentioning

confidence: 99%

NOX4 Inhibition Potentiates Immunotherapy by Overcoming Cancer-Associated Fibroblast-Mediated CD8 T-cell Exclusion from Tumors

et al. 2020

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Determining mechanisms of resistance to aPD-1/PD-L1 immune-checkpoint immunotherapy is key to developing new treatment strategies. Cancer-associated fibroblasts (CAF) have many tumor-promoting functions and promote immune evasion through multiple mechanisms, but as yet, no CAF-specific inhibitors are clinically available. Here we generated CAF-rich murine tumor models (TC1, MC38, and 4T1) to investigate how CAFs influence the immune microenvironment and affect response to different immunotherapy modalities [anticancer vaccination, TC1 (HPV E7 DNA vaccine), aPD-1, and MC38] and found that CAFs broadly suppressed response by specifically excluding CD8 þ T cells from tumors (not CD4 þ T cells or macrophages); CD8 þ T-cell exclusion was similarly present in CAF-rich human tumors. RNA sequencing of CD8 þ T cells from CAF-rich murine tumors and immunochemistry analysis of human tumors identified significant upregulation of CTLA-4 in the absence of other exhaustion markers; inhibiting CTLA-4 with a nondepleting antibody overcame the CD8 þ T-cell exclusion effect without affecting Tregs. We then examined the potential for CAF targeting, focusing on the ROS-producing enzyme NOX4, which is upregulated by CAF in many human cancers, and compared this with TGFb1 inhibition, a key regulator of the CAF phenotype. siRNA knockdown or pharmacologic inhibition [GKT137831 (Setanaxib)] of NOX4 "normalized" CAF to a quiescent phenotype and promoted intratumoral CD8 þ T-cell infiltration, overcoming the exclusion effect; TGFb1 inhibition could prevent, but not reverse, CAF differentiation. Finally, NOX4 inhibition restored immunotherapy response in CAFrich tumors. These findings demonstrate that CAF-mediated immunotherapy resistance can be effectively overcome through NOX4 inhibition and could improve outcome in a broad range of cancers. Significance: NOX4 is critical for maintaining the immunesuppressive CAF phenotype in tumors. Pharmacologic inhibition of NOX4 potentiates immunotherapy by overcoming CAFmediated CD8 þ T-cell exclusion.

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Section: Murine Tumor Modelsmentioning

confidence: 99%

NOX4 Inhibition Potentiates Immunotherapy by Overcoming Cancer-Associated Fibroblast-Mediated CD8 T-cell Exclusion from Tumors

et al. 2020

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“…The double-stranded structure of the DNA plasmid is also thought to be an immune stimulant [48] through non-TLR mechanisms. In fact, plasmid DNA also acts on the TBK1-STING pathway through cytosolic receptors [49,50]. This results in the generation of Type 1 interferons, which then act as adjuvants for the generation of immune responses against the antigen(s) encoded by the plasmid DNA vaccine.…”

Section: Inflammatory Responses and Toxicitiesmentioning

confidence: 99%

A Comparison of Plasmid DNA and mRNA as Vaccine Technologies

Liu¹

2019

Vaccines

310

297

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This review provides a comparison of the theoretical issues and experimental findings for plasmid DNA and mRNA vaccine technologies. While both have been under development since the 1990s, in recent years, significant excitement has turned to mRNA despite the licensure of several veterinary DNA vaccines. Both have required efforts to increase their potency either via manipulating the plasmid DNA and the mRNA directly or through the addition of adjuvants or immunomodulators as well as delivery systems and formulations. The greater inherent inflammatory nature of the mRNA vaccines is discussed for both its potential immunological utility for vaccines and for the potential toxicity. The status of the clinical trials of mRNA vaccines is described along with a comparison to DNA vaccines, specifically the immunogenicity of both licensed veterinary DNA vaccines and select DNA vaccine candidates in human clinical trials.

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“…The in vitro amplification technology produces DNA containing no bacterial origin of replication sequences or antibiotic selection marker. These vectors are capable of immunizing against influenza infection, with a response comparable to that of plasmid DNA [68], as well as improving tumor growth control in a Human papillomavirus (HPV) driven head and neck cancer model by delivering a therapeutic vaccine encoding for HPV16 E6 and E7 antigens [69]. Furthermore, dbDNA constructs have been used to generate functional lentiviral vectors [14] and are promising candidates for the production of recombinant adeno-associated virus (AAV) vectors [13].…”

Section: Applications In Biotechnologymentioning

confidence: 99%

The Unusual Linear Plasmid Generating Systems of Prokaryotes

Knott¹,

Rothwell²

2020

Bacteriophages - Perspectives and Future

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Linear DNA is vulnerable to exonuclease degradation and suffers from genetic loss due to the end replication problem. Eukaryotes overcome these problems by locating repetitive telomere sequences at the end of each chromosome. In humans and other vertebrates this noncoding terminal sequence is repeated between hundreds and thousands of times, ensuring important genetic information is protected. In most prokaryotes, the end-replication problem is solved by utilizing circular DNA molecules as chromosomes. However, some phage and bacteria do store genetic information in linear constructs, and the ends of these structures form either invertrons or hairpin telomeres. Hairpin telomere formation is catalyzed by a protelomerase, a unique protein that modifies DNA by a two-step transesterification reaction, proceeding via a covalent protein bound intermediate. The specifics of this mechanism are largely unknown and conflicting data suggests variations occur between different systems. These proteins, and the DNA constructs they produce, have valuable applications in the biotechnology industry. They are also an essential component of some human pathogens, an increased understanding of how they operate is therefore of fundamental importance. Although this review will focus on phage encoded protelomerase, protelomerases found from Agrobacterium and Borellia will be discussed in terms of mechanism of action.

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Linear doggybone DNA vaccine induces similar immunological responses to conventional plasmid DNA independently of immune recognition by TLR9 in a pre-clinical model

Cited by 31 publications

References 45 publications

NOX4 Inhibition Potentiates Immunotherapy by Overcoming Cancer-Associated Fibroblast-Mediated CD8 T-cell Exclusion from Tumors

NOX4 Inhibition Potentiates Immunotherapy by Overcoming Cancer-Associated Fibroblast-Mediated CD8 T-cell Exclusion from Tumors

A Comparison of Plasmid DNA and mRNA as Vaccine Technologies

The Unusual Linear Plasmid Generating Systems of Prokaryotes

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