DNA Vaccine: A Defense against Bioterrorism?

Gema Puspa Sari

Budiman BelaVirology and Cancer Pathobiology Research Center for

Health Service (VCPRC) FKUI-RSCM

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Introduction

• DNA Vaccine

– Using genetically engineered DNA so cells directly produce an antigens, resulting protective immunological response

• Advantages:

– Can be prepared from small amounts of DNA isolated from a pathogen; no need to culture or attenuate large stock of deadly bacteria or viruses

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DNA Vaccines

• Bioterrorism

– Terrorism involving the intentional release or dissemination of biological agent

– Bacteria, virus, toxin; natural or modified

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Distribution: up to 2,5 months

Packaging, transportation

Control agency testing: up to 2 months

Testing further toxicology, efficacy and clinical trial testing

Product finishing: 1-2,5 months

Production of ampoules, vials or syringes from bulk, testing and correct volumes

Quality control: up to 3 months

Single bulk tested: identity, potency, efficacy, purity, sterility, pH composition, or buffer, etc.

Production: 4-9 months

Preparation of raw materials, cultivation of viruses or bacteria, production of a single bulk

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(Forde, 2005)

Distribution: up to 2,5 months

Packaging, transportation

Control agency testing: up to 2 months

Testing further toxicology, efficacy and clinical trial testing

Product finishing: 1-2,5 months

Production of ampoules, vials or syringes from bulk, testing and correct volumes

Quality control: up to 3 months

Single bulk tested: identity, potency, efficacy, purity, sterility, pH composition, or buffer, etc.

Production: 4-9 months

Preparation of raw materials, cultivation of viruses or bacteria, production of a single bulk

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1 month

(Forde, 2005)

• Foreign DNA (from bacteria, parasites, yeast, and insects) may induce a protective innate immune response– Immunostimulatory oligodeoxynucleotides containing

CpG motif (CpG ODN) -> able to induce cytokines and activate B cells, monocyte, dendritic cell, and NK cells

– CpG ODN (CpG oligodeoxynucleotide) is short single-stranded synthetic DNA molecule that contain a cytosine triphosphate deoxynucleotide (‘C’) followed by a guanine triphosphate deoxynucleotid (‘G’), ‘p’ refers to phosphodiester linked between them.

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• Advantages of CpG ODN:

– Boost the immune response -> higher IFN-y, antigen specific CTL

– Enhance mucosal immunity

– Boost immunity in group with reduce immune function

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PATHOGEN

DNA Vaccine CpG ODN

B

Th

CTL

B

NK

MФ,

DC

T

ProliferationMemory

IFN-y ; IL-4

ProliferationMemory

Ag-specificIgG

Ag-specificcytotoxicity

IL-6, 12, proliferation

Activation IFN α, β, γ

IL-6, 12, 18, TNFα,

activation,

maturation

Activation,proliferation

Direct

Indirect

Polyreactive IgM

NKkilling

phagocytosis

Killing

(Klinman et al, 1999)

Mechanism of Protection

Examples

• Ebola and Yersinia Vaccine, targeting:

– Cell curface (Vanderzanden et al, 1998)

– Envelope (Xu et al, 1998)

– Nucleoprotein (Noll, 1999)

• Anthrax -> pathogenic agent component from anthrax toxin (Gu et al, 1999)

• Integrating into the host genome

• Inducing autoimmune disease

• Altering immune homeostatsis

• Toxic

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Risks

Conclusion• DNA vaccine strategy has potential to

contribute in short-term and persistent protection from the threat of bioterrorism.

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References

Ashkar AA, Rosenthal KL. Toll-like receptor 9, CpG DNA and innate immunity. Current Molecular Medicine 2002, 2, 545-56

Forde GM. 2005. Rapid-response vaccines—does DNA offer a solution? Nature Biotechnology 23; 9; 1059-62

Klinman DM, Verthelyi D, Takeshitas F, et al. 1999. Immune recognition of foreign DNA: A cure for bioterrorism? Immunity 11, 123-9.

Krieg AM. Immune effects and mechanism of action of CpG motifs. Vaccine 19 (2001) 618–622

Ulmer JB, Donnelly JJ, Parker SE, et al. 1993. Heterologous protection against influenza by injection of DNA encoding a viral protection. Science 259, 1745-49.

Weiner GJ, Liu HM, Wooldridge JE, et al. 1997. Immunostimulatory oligodeoxynucloetides containing the CpG motif are effective as immune adjuvants in tumor antigen immunization. Proc Natl Acad Sci USA 94, 10833-7.

DNA Vaccine: A Defense against Bioterrorism?Gema Puspa Sari, Budiman Bela

Virology and Cancer Pathobiology Research Center for Health Service FKUI-RSCMCorresponding author: gemapuspasari@yahoo.com

AbstractBiological substances have a potential to be used as biological weapon in terrorism. To counter bioterrorism threat,researchers are developing many strategies to boosting the host’s resistance to infectious agent. DNA vaccinescomposed gene that coding the antigen whose expression is regulated by mammalian promoter which are expressedon a plasmid backbone of bacterial DNA.1 One of several advantages in using DNA plasmids over conventional vaccine isit can be prepared from small amount of DNA isolated from the pathogen without needing to culture or attenuate thepathogen source such as bacteria or virus. Futhermore, DNA vaccines capable to induce innate and adaptive immuneresponse to counter biothreat agents in bioterrorism.2 DNA vaccines have immunostimulatory oligodeoxynucleotidescontaining CpG motif (CpG ODN) which able to induce cytokines and activate B cells, monocyte, dendritic cell, and NKcells.3 Combining immunomodulatory properties of CpG ODN with the antigen-specific responses induced by DNAvaccine can be a strategy to reduce the threat from bioterrorism.2

ConclusionDNA vaccine strategy has potential to contribute in short-term and persistent protection from the threat of bioterrorism.

Keywords: DNA vaccine, CpG ODN, immune response, bioterrorism, biothreat

References 1. Ulmer JB, Donnelly JJ, Parker SE, et al. 1993. Heterologous protection against influenza by injection of DNA

encoding a viral protection. Science 259, 1745-49.2. Klinman DM, Verthelyi D, Takeshitas F, et al. 1999. Immune recognition of foreign DNA: A cure for bioterrorism?

Immunity 11, 123-9.3. Weiner GJ, Liu HM, Wooldridge JE, et al. 1997. Immunostimulatory oligodeoxynucloetides containing the CpG motif

are effective as immune adjuvants in tumor antigen immunization. Proc Natl Acad Sci USA 94, 10833-7.