scaricalo in formato PDF - labogen srl

ANTIBODIES & VACCINATION 5
pBOOST - DNA Vaccine Adjuvants
pBOOST plasmids were developed as genetic adjuvants for DNA vaccines to potentiate the immune response to a specific antigen. The method
of plasmid DNA vaccine delivery is known to bias the immune response to a specific antigen towards aTh1 or Th2 response. These biases can
be further enhanced by the codelivery of interferon regulatory factors (IRFs) orTANK-binding kinase 1 (TBK1) to increase the efficacy of the
vaccination.
Description
pBOOST2-mIRF - Th1 or Th2 polarization
IRF-1, IRF-3 and IRF-7 have been shown to be able to bias T cells towards
type 1 or type 2 immune responses, leading to the activation of cytotoxic
T cells and/or the production of antibodies.
pBOOST2-mIRF plasmids carry the murine IRF-1, IRF-3 or IRF-7 genes.
These genes are either wild-type or mutant.
• Wild-type IRF-1: IRF-1 primarily increases Th2 antibody responses 1 .
Following intramuscular or gene gun injections of DNA vaccines, IRF-1 can
increase the titers of antibodies up to 10-fold.
• Mutant IRF-3: IRF-3 primarily increases Th1 T-cell responses 1 . A
constitutively active form of IRF-3 was generated by creating a single point
mutation of Ser 396 to Asp. This super-activated IRF-3 presents a >10-fold
enhanced transactivating potential over the wild-type IRF-3 for the IFN-a
and IFN-b promoters 2 .
• Chimeric IRF7/3: IRF-3 and IRF-7 increase both Th1 and Th2 responses
by transactivating different target promoters 1 . To exploit the biological
features of both IRFs, a chimeric form of IRF-7 and IRF-3 was generated
by combining the DNA binding specificity of IRF-7 with the strong
transactivation capacity of super-activated IRF-3. IRF-7/3 chimera provides
>10-fold greater induction of IFN-a and IFN-b promoters than superactivated
IRF-3 alone 3 .
pBOOST2-mIRF plasmids express the transgene under the control of the
strong and ubiquitous EF-1a/HTLV composite promoter and are selectable
in E. coli with Zeocin .
pBOOST3-mTBK1 - Enhancement of DNA vaccine immunogenicity
pBOOST3-mTBK1 plasmid expresses the mouse TBK1 gene. TANKbinding
kinase 1 (TBK1), a non-canonical IkB kinase, was recently shown
to mediate the adjuvant effect of DNA vaccines 4 . Administration of DNA
vaccines induces the production of type I interferons and inflammatory
cytokines in a CpG-independent manner but in a TBK1-dependent manner.
Therefore, co-administration of a TBK1-expressing plasmid is expected to
further boost DNA vaccine-induced immunogenicity.
pBOOST3-mTBK1 contains the EF-1a/HTLV composite promoter
coupled to the SV40 enhancer and is selectable in E. coli with Zeocin .
1. Sasaki S. et al., 2002. Regulation of DNA-raised immune responses by cotransfected
interferon regulatory factors. J Virol.76(13):6652-9. 2. Servant MJ. et al., 2003.
Identification of the minimal phosphoacceptor site required for in vivo activation of
interferon regulatory factor 3 in response to virus and double-stranded RNA. J Biol
Chem. 278(11):9441-7. 3. Bramson JL. et al., 2003. Super-activated interferon-regulatory
factors can enhance plasmid immunization. Vaccine. 21(13-14):1363-70. 4. Ishii KJ. et al.,
2008. TANK-binding kinase-1 delineates innate and adaptive immune responses to DNA
vaccines. Nature 451: 725-729
Contents and Storage
130 www.invivogen.com/dna-vaccination
Each pBOOST plasmid is provided as 20 μg of lyophilized DNA. Product
is shipped at room temperature. Store at -20°C. Plasmids are stable up to
one year when properly stored. pBOOST plasmids come with 4 pouches
of E. coli Fast-Media ® Zeo (2 TB and 2 Agar, see pages 44-45).
PRODUCT QTY CAT. CODE
pBOOST2-wtmIRF1 20 μg pbst2-wtmirf1
pBOOST2-samIRF3 20 μg pbst2-samirf3
pBOOST2-samIRF7/3 20 μg pbst2-samirf73
pBOOST2-mcs (control) 20 μg pbst2-mcs
pBOOST3-mTBK1 20 μg pbst3-mtbk1
pBOOST3-mcs (control) 20 μg pbst3-mcs