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Poly(I:C)
Synthetic double-stranded RNA, namely poly(I:C), can activate the immune response through two distinct PRRs 2 . Endosomal poly(I:C) activates TLR3 while
cytosolic poly(I:C) activates RIG-I/MDA-5. Triggering the TLR3 pathway induces IL-12 and type I IFNs production, and improves MHC class II expression and
cross-presentation of antigen 2 . Stimulation of MDA-5 enhances the production of type I IFNs that play a critical role in enhancing T and B cell immunity 2 .
Poly(I:C) promotes Th1 biased immunity through its induction of IL-12 and type I IFNs 2 . Promising results have been obtained using poly(I:C) as an adjuvant
in flu vaccine delivered intranasally 9 .
MPLA
The TLR4 agonist, MPLA (monophosphoryl lipid A) is a derivative of lipid A from Salmonella minnesota R595 lipopolysaccharide (LPS or endotoxin). MPLA
is considerably less toxic than LPS whilst maintaining the immunostimulatory activity 10 . When tested in animals models as a vaccine adjuvant, MPLA induces
a strong Th1 response 11 . The adjuvant, AS04, currently approved for use in vaccines against human papilloma virus and hepatitis B, contains MPL (a modified
MPLA) combined with aluminum salt 12 .
N-glycolyl-MDP
MDP (muramyl dipeptide), is the minimal bioactive peptidoglycan motif common to all bacteria and the essential structure required for adjuvant activity in
vaccines. MDP has been shown to be recognized by NOD2, but not TLR2, nor TLR2/1 or TLR2/6 associations 13 . The cell wall of mycobacteria is known to
be extremely immunogenic. This potent activity is attributed to their MDP which is N-glycolylated in contrast to the MDP of most bacteria which is
N-acetylated. N-glycolyl-MDP has been reported to display a stronger NOD2-mediated activity than N-acetyl-MDP and thus to be a more potent vaccine
adjuvant than N-acetyl-MDP 14 . Furthermore MDP leads to the activation of the NLRP3 inflammasome 15 .
ODN 1826 and ODN 2006
Synthetic oligodeoxynucleotides containing unmethylated CpG motifs (CpG ODNs), such as ODN 2006 (also known as ODN 7909), have been extensively
studied as adjuvants. CpG ODNs are recognized by TLR9, which is expressed exclusively on human B cells and plasmacytoid dendritic cells (pDCs), thereby
inducing Th1-dominated immune responses. Pre-clinical studies conducted in rodents and non-human primates and human clinical trials have demonstrated that
CpG ODNs can significantly improve vaccine-specific antibody responses 9 . Among the three classes of CpG ODNs identified (see page 86), type B CpG ODNs
are the most studied.
1. Mbow ML. et al., 2010. New adjuvants for human vaccines. Curr Opin Immunol. 22(3):411-6. 2. Coffman RL. et al., 2010. Vaccine adjuvants: Putting innate immunity to work. Immunity
33(4):492-503. 3. Marrack P. et al., 2009. Towards an understanding of adjuvant action of aluminium. Nat Rev Immunol. 9(4): 287-93. 4. Petrovsky N. & Aguilar JC., 2004. Vaccine adjuvants:
Current state and future trends. Immunol Cell Biol. 82(5): 488-96. 5. Moreira L0. et al., 2008. Modulation of adaptive immunity by different adjuvant-antigen combinations in mice lacking
Nod2. Vaccine 26(46): 5808-13. 6. Huleatt J. et al., 2007. Vaccination with recombinant fusion proteins incorporating Toll-like receptor ligands induces rapid cellular and humoral immunity.
Vaccine 25(4): 763-75. 7. Skountzou I. et al., 2010. Salmonella flagellins are potent adjuvants for intranasally administered whole inactivated influenza vaccine. Vaccine 28(4): 4103-12. 8. Miao
EA. & Warren SE., 2010. Innate immune detection of bacterial virulence factors via the NLRC4 inflammasome. J Clin Immunol. 30(4): 502-6. 9. Steinhagen F. et al., 2010. TLR-based immune
adjuvants. Vaccine [Epub ahead of print]. 10. Casella CR. et al., 2008. Putting endotoxin to work for us: monophosphoryl lipid A as a safe and effective vaccine adjuvant. Cell Mol Life Sci. 65(20):3231-
40. 11. Fransen F. et al., 2007. Agonists of Toll-like receptors 3, 4, 7, and 9 are candidates for use as adjuvants in an outer membrane vaccine against Neisseria meningitidis serogroup . Infect Immun.
75(12) :5939-46. 12. Didierlaurent A. et al., 2009. AS04, an aluminum salt- and TLR4 agonist-based adjuvant system, induces a transient localized innate immune response leading to enhanced
adaptive immunity. J Immunol 183(10): 6186-97. 13. Girardin S. et al., 2003. Nod2 is a general sensor of peptidoglycan through muramyl dipeptide (MDP) detection. J Biol Chem. 278(11):8869-
72. 14. Coulombe F. et al., 2009. Increased NOD2-mediated recognition of N-glycolyl muramyl dipeptide. J Exp Med. 206(8):1709-16. 15. Martinon F. et al., 2004. Identification of bacterial
muramyl dipeptide as activator of the NALP3/cryopyrin inflammasome. Curr Biol 14 (21): 1929-34.
OVA Antigens NEW
Ovalbumin (OVA) is a key reference protein for immunization and biochemical studies (Western blots, ELISA). Commercially available ovalbumin
is often contaminated with endotoxins altering the results in vivo. InvivoGen provides two grades of ovalbumin and two standard OVA peptides.
• EndoFit Ovalbumin: Endotoxin level < 1 EU/mg, for in vivo use,
minimum 98% protein
• Ovalbumin: for detection use (Western blot, ELISA), minimum 98%
protein
• OVA 257-264: an H-2Kb-restricted OVA class I epitope, for
detection use (ELISPOT) - Sequence : SIINFEKL (MW 963.2)
• OVA 323-339: an H-2b-restricted OVA class II epitope, for detection
use (ELISPOT) - Sequence : ISQAVHAAHAEINEAGR (MW 1773.9)
Contents and Storage
Products are provided lyophilized and shipped at room temperature.
Store at 4°C or -20°C according to the product label.
PRODUCT QTY CAT. CODE
EndoFit Ovalbumin 10 mg vac-efova
Ovalbumin 1 g vac-ova
OVA 257-264 1 mg vac-sin
OVA 323-339 1 mg vac-isq
www.invivogen.com/vaccine-adjuvants 129
ANTIBODIES & VACCINATION 5