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The Carbomer-Lecithin Adjuvant Adjuplex Has Potent Immunoactivating Properties and Elicits Protective Adaptive Immunity against Influenza Virus Challenge in Mice.

Wegmann F, Moghaddam AE, Schiffner T, Gartlan KH, Powell TJ, Russell RA, Baart M, Carrow EW, Sattentau QJ - Clin. Vaccine Immunol. (2015)

Bottom Line: The continued discovery and development of adjuvants for vaccine formulation are important to safely increase potency and/or reduce the antigen doses of existing vaccines and tailor the adaptive immune response to newly developed vaccines.Adjuplex neither triggered classical maturation of DCs nor activated a pathogen recognition receptor (PRR)-expressing NF-κB reporter cell line, suggesting a mechanism of action different from that reported for classical pathogen-associated molecular pattern (PAMP)-activated innate immunity.Taken together, these data reveal Adjuplex to be a potent and well-tolerated adjuvant with application for subunit vaccines.

View Article: PubMed Central - PubMed

Affiliation: The Sir William Dunn School of Pathology, The University of Oxford, Oxford, United Kingdom Janssen Infectious Diseases and Vaccines, Leiden, The Netherlands.

No MeSH data available.


Related in: MedlinePlus

Lack of innate immune cell and receptor triggering by Adjuplex. (a) Bone marrow-derived dendritic cells (BMDCs) were incubated with the concentrations of Adjuplex or the PRR ligands indicated, and surface marker expression was analyzed by flow cytometry and expressed as the percentage of positively stained cells compared to isotype controls. (b) Thp1-Blue reporter cells were pulsed with the compounds shown for 24 h, and the supernatants were assayed for secreted embryonic alkaline phosphatase. Data represent the mean ± SEM. *, P < 0.05; ***, P < 0.001; n.s., not significant; OD630nm, optical density at 630 nm.
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Figure 7: Lack of innate immune cell and receptor triggering by Adjuplex. (a) Bone marrow-derived dendritic cells (BMDCs) were incubated with the concentrations of Adjuplex or the PRR ligands indicated, and surface marker expression was analyzed by flow cytometry and expressed as the percentage of positively stained cells compared to isotype controls. (b) Thp1-Blue reporter cells were pulsed with the compounds shown for 24 h, and the supernatants were assayed for secreted embryonic alkaline phosphatase. Data represent the mean ± SEM. *, P < 0.05; ***, P < 0.001; n.s., not significant; OD630nm, optical density at 630 nm.

Mentions: Adjuvants that bind PRRs trigger NF-κB activation pathways, leading to the maturation of immature DCs and upregulation of costimulatory surface molecules that allow efficient antigen-specific priming of T cells. We therefore tested whether Adjuplex might represent a PRR ligand by pulsing DCs with two concentrations of Adjuplex and reading out surface HLA and costimulatory marker expression. The highest dose (5%) of Adjuplex generated a high fluorescent background in the BMDCs and so could not be used for the analysis. In contrast with lipopolysaccharide (LPS), which upregulated MHC-II, CD80, CD86, CD40, and OX40L expression, there was no significant increase in any surface markers in the presence of Adjuplex (Fig. 7a). To confirm that Adjuplex was unable to directly trigger PRRs, we pulsed an NF-κB myeloid reporter cell line, Thp1-Blue, expressing many functional PRRs, including TLR1/2, TLR2, TLR2/6, TLR4, TLR5, TLR8, NOD1, and NOD2, with a range of Adjuplex concentrations and read out NF-κB activation. Whereas the known ligands LPS (TLR4), lipoteichoic acid from S. aureus (LTA-SA) (TLR2), FSL-1 (TLR2/6), and muramyl dipeptide (MDP) (NOD2) all activated NF-κB, none of the 3 concentrations of Adjuplex induced any significant activation (Fig. 7b). Taken together with the lack of canonical DC maturation, we therefore conclude that the major adjuvant activity of Adjuplex is unlikely to depend upon direct DC activation via PRRs.


The Carbomer-Lecithin Adjuvant Adjuplex Has Potent Immunoactivating Properties and Elicits Protective Adaptive Immunity against Influenza Virus Challenge in Mice.

Wegmann F, Moghaddam AE, Schiffner T, Gartlan KH, Powell TJ, Russell RA, Baart M, Carrow EW, Sattentau QJ - Clin. Vaccine Immunol. (2015)

Lack of innate immune cell and receptor triggering by Adjuplex. (a) Bone marrow-derived dendritic cells (BMDCs) were incubated with the concentrations of Adjuplex or the PRR ligands indicated, and surface marker expression was analyzed by flow cytometry and expressed as the percentage of positively stained cells compared to isotype controls. (b) Thp1-Blue reporter cells were pulsed with the compounds shown for 24 h, and the supernatants were assayed for secreted embryonic alkaline phosphatase. Data represent the mean ± SEM. *, P < 0.05; ***, P < 0.001; n.s., not significant; OD630nm, optical density at 630 nm.
© Copyright Policy - open-access
Related In: Results  -  Collection

License
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getmorefigures.php?uid=PMC4550664&req=5

Figure 7: Lack of innate immune cell and receptor triggering by Adjuplex. (a) Bone marrow-derived dendritic cells (BMDCs) were incubated with the concentrations of Adjuplex or the PRR ligands indicated, and surface marker expression was analyzed by flow cytometry and expressed as the percentage of positively stained cells compared to isotype controls. (b) Thp1-Blue reporter cells were pulsed with the compounds shown for 24 h, and the supernatants were assayed for secreted embryonic alkaline phosphatase. Data represent the mean ± SEM. *, P < 0.05; ***, P < 0.001; n.s., not significant; OD630nm, optical density at 630 nm.
Mentions: Adjuvants that bind PRRs trigger NF-κB activation pathways, leading to the maturation of immature DCs and upregulation of costimulatory surface molecules that allow efficient antigen-specific priming of T cells. We therefore tested whether Adjuplex might represent a PRR ligand by pulsing DCs with two concentrations of Adjuplex and reading out surface HLA and costimulatory marker expression. The highest dose (5%) of Adjuplex generated a high fluorescent background in the BMDCs and so could not be used for the analysis. In contrast with lipopolysaccharide (LPS), which upregulated MHC-II, CD80, CD86, CD40, and OX40L expression, there was no significant increase in any surface markers in the presence of Adjuplex (Fig. 7a). To confirm that Adjuplex was unable to directly trigger PRRs, we pulsed an NF-κB myeloid reporter cell line, Thp1-Blue, expressing many functional PRRs, including TLR1/2, TLR2, TLR2/6, TLR4, TLR5, TLR8, NOD1, and NOD2, with a range of Adjuplex concentrations and read out NF-κB activation. Whereas the known ligands LPS (TLR4), lipoteichoic acid from S. aureus (LTA-SA) (TLR2), FSL-1 (TLR2/6), and muramyl dipeptide (MDP) (NOD2) all activated NF-κB, none of the 3 concentrations of Adjuplex induced any significant activation (Fig. 7b). Taken together with the lack of canonical DC maturation, we therefore conclude that the major adjuvant activity of Adjuplex is unlikely to depend upon direct DC activation via PRRs.

Bottom Line: The continued discovery and development of adjuvants for vaccine formulation are important to safely increase potency and/or reduce the antigen doses of existing vaccines and tailor the adaptive immune response to newly developed vaccines.Adjuplex neither triggered classical maturation of DCs nor activated a pathogen recognition receptor (PRR)-expressing NF-κB reporter cell line, suggesting a mechanism of action different from that reported for classical pathogen-associated molecular pattern (PAMP)-activated innate immunity.Taken together, these data reveal Adjuplex to be a potent and well-tolerated adjuvant with application for subunit vaccines.

View Article: PubMed Central - PubMed

Affiliation: The Sir William Dunn School of Pathology, The University of Oxford, Oxford, United Kingdom Janssen Infectious Diseases and Vaccines, Leiden, The Netherlands.

No MeSH data available.


Related in: MedlinePlus