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Immunity induced by a broad class of inorganic crystalline materials is directly controlled by their chemistry.

Williams GR, Fierens K, Preston SG, Lunn D, Rysnik O, De Prijck S, Kool M, Buckley HC, Lambrecht BN, O'Hare D, Austyn JM - J. Exp. Med. (2014)

Bottom Line: Using a systems vaccinology approach, we find that every measured response can be modeled using a subset of just three physical and chemical properties for all compounds tested.This correlation can be reduced to a simple linear equation that enables the immunological responses stimulated by newly synthesized LDHs to be predicted in advance from these three parameters alone.This study demonstrates that immunity can be determined purely by chemistry and opens the possibility of rational manipulation of immunity for therapeutic purposes.

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Affiliation: Chemistry Research Laboratory, Department of Chemistry; Nuffield Department of Surgical Sciences, John Radcliffe Hospital; and Department of Statistics; University of Oxford, Oxford OX1 2JD, England, UKChemistry Research Laboratory, Department of Chemistry; Nuffield Department of Surgical Sciences, John Radcliffe Hospital; and Department of Statistics; University of Oxford, Oxford OX1 2JD, England, UK.

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Chemically different LDHs drive diverse DC responses in vitro. Human monocyte-derived DCs were cultured without or with the indicated LDH or commercial adjuvant preparation for a period of 24 h. The concentration of cytokines and chemokines in the supernatant was then determined using ELISA (IL-6 and TNF) or Luminex (others). Cell surface expression of co-stimulatory and co-inhibitory molecules was assessed by flow cytometry. MFI, mean fluorescence intensity. Error bars show one standard error. *, P < 0.05; and **, P < 0.01 versus cells alone. The number of individual experiments performed for each response ranges from n = 6 to n = 22 and is given in Table S3. Each experiment contained at least three biological replicates.
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fig2: Chemically different LDHs drive diverse DC responses in vitro. Human monocyte-derived DCs were cultured without or with the indicated LDH or commercial adjuvant preparation for a period of 24 h. The concentration of cytokines and chemokines in the supernatant was then determined using ELISA (IL-6 and TNF) or Luminex (others). Cell surface expression of co-stimulatory and co-inhibitory molecules was assessed by flow cytometry. MFI, mean fluorescence intensity. Error bars show one standard error. *, P < 0.05; and **, P < 0.01 versus cells alone. The number of individual experiments performed for each response ranges from n = 6 to n = 22 and is given in Table S3. Each experiment contained at least three biological replicates.

Mentions: Because alum adjuvants act on DCs, at least in vivo (Kool et al., 2008a,b; Marrack et al., 2009; McKee et al., 2009), we initially evaluated the immunological properties of LDHs in culture by assessing their ability to stimulate responses of human monocyte-derived DCs (Sallusto and Lanzavecchia, 1994). We measured the capacity of DCs to secrete proinflammatory cytokines (IL-1β, IL-6, and TNF), as well as cytokines that polarize Th1 responses (IL-12p70), maintain memory T cell responses (IL-15), and promote antiviral responses (IFN-α2). We also investigated DC production of chemokines (IL-8, MIP-1β, and MCP-1) and of membrane molecules involved in T cell activation (CD86), DC–T cell cross talk (CD40), and suppression of CD8 T cell proliferation (CD274). To study this systematically, we designed, synthesized, and characterized a series of endotoxin-free LDHs, with systematic substitutions of metal cations and anions (Table S1). The LDHs were observed to comprise crystalline hexagonal platelets (Fig. 1). DCs were co-cultured with LDHs, and the cellular responses stimulated by each compound were evaluated and compared with those elicited by the commercially available alums Imject and Alhydrogel. We found that individual LDHs stimulated distinct DC responses across many immunological outputs but with donor to donor variation. We therefore undertook a statistical repeated measures analysis, with a random effect to account for donor variability, and fitted a multilevel model to obtain estimates of the overall responses. Several compounds were found to stimulate robust DC responses, frequently much greater than the commercially used Alhydrogel adjuvant (Fig. 2 and Tables S2 and S3). In Fig. 2 and subsequently, LDHs are denoted MxMy′-X, where M and M′ are the metal cations in the LDH layers, x/y is the ratio of these ions, and X is the interlayer anion.


Immunity induced by a broad class of inorganic crystalline materials is directly controlled by their chemistry.

Williams GR, Fierens K, Preston SG, Lunn D, Rysnik O, De Prijck S, Kool M, Buckley HC, Lambrecht BN, O'Hare D, Austyn JM - J. Exp. Med. (2014)

Chemically different LDHs drive diverse DC responses in vitro. Human monocyte-derived DCs were cultured without or with the indicated LDH or commercial adjuvant preparation for a period of 24 h. The concentration of cytokines and chemokines in the supernatant was then determined using ELISA (IL-6 and TNF) or Luminex (others). Cell surface expression of co-stimulatory and co-inhibitory molecules was assessed by flow cytometry. MFI, mean fluorescence intensity. Error bars show one standard error. *, P < 0.05; and **, P < 0.01 versus cells alone. The number of individual experiments performed for each response ranges from n = 6 to n = 22 and is given in Table S3. Each experiment contained at least three biological replicates.
© Copyright Policy - openaccess
Related In: Results  -  Collection

License 1 - License 2
Show All Figures
getmorefigures.php?uid=PMC4042647&req=5

fig2: Chemically different LDHs drive diverse DC responses in vitro. Human monocyte-derived DCs were cultured without or with the indicated LDH or commercial adjuvant preparation for a period of 24 h. The concentration of cytokines and chemokines in the supernatant was then determined using ELISA (IL-6 and TNF) or Luminex (others). Cell surface expression of co-stimulatory and co-inhibitory molecules was assessed by flow cytometry. MFI, mean fluorescence intensity. Error bars show one standard error. *, P < 0.05; and **, P < 0.01 versus cells alone. The number of individual experiments performed for each response ranges from n = 6 to n = 22 and is given in Table S3. Each experiment contained at least three biological replicates.
Mentions: Because alum adjuvants act on DCs, at least in vivo (Kool et al., 2008a,b; Marrack et al., 2009; McKee et al., 2009), we initially evaluated the immunological properties of LDHs in culture by assessing their ability to stimulate responses of human monocyte-derived DCs (Sallusto and Lanzavecchia, 1994). We measured the capacity of DCs to secrete proinflammatory cytokines (IL-1β, IL-6, and TNF), as well as cytokines that polarize Th1 responses (IL-12p70), maintain memory T cell responses (IL-15), and promote antiviral responses (IFN-α2). We also investigated DC production of chemokines (IL-8, MIP-1β, and MCP-1) and of membrane molecules involved in T cell activation (CD86), DC–T cell cross talk (CD40), and suppression of CD8 T cell proliferation (CD274). To study this systematically, we designed, synthesized, and characterized a series of endotoxin-free LDHs, with systematic substitutions of metal cations and anions (Table S1). The LDHs were observed to comprise crystalline hexagonal platelets (Fig. 1). DCs were co-cultured with LDHs, and the cellular responses stimulated by each compound were evaluated and compared with those elicited by the commercially available alums Imject and Alhydrogel. We found that individual LDHs stimulated distinct DC responses across many immunological outputs but with donor to donor variation. We therefore undertook a statistical repeated measures analysis, with a random effect to account for donor variability, and fitted a multilevel model to obtain estimates of the overall responses. Several compounds were found to stimulate robust DC responses, frequently much greater than the commercially used Alhydrogel adjuvant (Fig. 2 and Tables S2 and S3). In Fig. 2 and subsequently, LDHs are denoted MxMy′-X, where M and M′ are the metal cations in the LDH layers, x/y is the ratio of these ions, and X is the interlayer anion.

Bottom Line: Using a systems vaccinology approach, we find that every measured response can be modeled using a subset of just three physical and chemical properties for all compounds tested.This correlation can be reduced to a simple linear equation that enables the immunological responses stimulated by newly synthesized LDHs to be predicted in advance from these three parameters alone.This study demonstrates that immunity can be determined purely by chemistry and opens the possibility of rational manipulation of immunity for therapeutic purposes.

View Article: PubMed Central - HTML - PubMed

Affiliation: Chemistry Research Laboratory, Department of Chemistry; Nuffield Department of Surgical Sciences, John Radcliffe Hospital; and Department of Statistics; University of Oxford, Oxford OX1 2JD, England, UKChemistry Research Laboratory, Department of Chemistry; Nuffield Department of Surgical Sciences, John Radcliffe Hospital; and Department of Statistics; University of Oxford, Oxford OX1 2JD, England, UK.

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