Limits...
Adsorption of Toll-Like Receptor 4 Agonist to Alum-Based Tetanus Toxoid Vaccine Dampens Pro-T Helper 2 Activities and Enhances Antibody Responses.

Bortolatto J, Mirotti L, Rodriguez D, Gomes E, Russo M - J Immunol Res (2015)

Bottom Line: Here we found that alum-based tetanus toxoid (TT) vaccine was biased toward a Th-2 profile as shown by TT-induced airway eosinophilic inflammation, type 2 cytokine production, and high levels of IgE anaphylactic antibodies.Importantly, in a situation with antigenic competition (OVA plus TT), TT-specific IgG1 or IgG2a was decreased compared with TT sensitization.In conclusion, the addition of LPS induces a more robust IgG1 and IgG2a TT-specific antibody production and concomitantly decreases Th2-cellular and humoral responses, indicating a potential use of alum/TLR-based vaccines.

View Article: PubMed Central - PubMed

Affiliation: Department of Immunology, Institute of Biomedical Science, University of São Paulo, 05508-000 São Paulo, SP, Brazil.

ABSTRACT
Aluminum salts gels (alum) are TLR-independent adjuvants and have been used to boost antibody responses in alum-based vaccines such as diphtheria, pertussis, and tetanus toxoid (DPT) triple vaccine. However, the pro-Th2 activity of alum-based vaccine formulations has not been fully appreciated. Here we found that alum-based tetanus toxoid (TT) vaccine was biased toward a Th-2 profile as shown by TT-induced airway eosinophilic inflammation, type 2 cytokine production, and high levels of IgE anaphylactic antibodies. The adsorption into alum of prototypic TLR4 agonists such as lipopolysaccharides (LPS) derived from Escherichia coli consistently dampened TT-induced Th2 activities without inducing IFNγ or Th1-like responses in the lung. Conversely, adsorption of monophosphoryl lipid A (MPLA) extracted from Salmonella minnesota, which is a TIR-domain-containing adapter-inducing interferon-β- (TRIF-) biased TLR4 agonist, was less effective in decreasing Th-2 responses. Importantly, in a situation with antigenic competition (OVA plus TT), TT-specific IgG1 or IgG2a was decreased compared with TT sensitization. Notably, LPS increased the production of IgG1 and IgG2a TT-specific antibodies. In conclusion, the addition of LPS induces a more robust IgG1 and IgG2a TT-specific antibody production and concomitantly decreases Th2-cellular and humoral responses, indicating a potential use of alum/TLR-based vaccines.

No MeSH data available.


Related in: MedlinePlus

Effects of adsorption of LPS or MPLA onto TT/alum sensitization on TT-mediated airway allergic responses. (a) Protocol: BALB/c mice sensitized with s.c. TT/alum in the presence or not of LPS (10 μg) or MPLA (10 μg) on days 0 and 7, and challenged with TT i.n. on days 14 and 21. Samples obtained on day 22: (b) total number of cells and (c) eosinophils in BAL; ((d) and (e)) levels of IL-5 and IL-13 in BAL. Data shown as mean ± SE, one-way ANOVA: #P < 0.05; different from TT/alum/PBS group (n = 5), and experiment was repeated twice.
© Copyright Policy - open-access
Related In: Results  -  Collection


getmorefigures.php?uid=PMC4562177&req=5

fig2: Effects of adsorption of LPS or MPLA onto TT/alum sensitization on TT-mediated airway allergic responses. (a) Protocol: BALB/c mice sensitized with s.c. TT/alum in the presence or not of LPS (10 μg) or MPLA (10 μg) on days 0 and 7, and challenged with TT i.n. on days 14 and 21. Samples obtained on day 22: (b) total number of cells and (c) eosinophils in BAL; ((d) and (e)) levels of IL-5 and IL-13 in BAL. Data shown as mean ± SE, one-way ANOVA: #P < 0.05; different from TT/alum/PBS group (n = 5), and experiment was repeated twice.

Mentions: We next adapted the OVA model protocol to tetanus toxoid antigen. As depicted in Figure 2(a), sensitizations of tetanus toxoid adsorbed to alum (TT/alum group) followed by i.n. challenges resulted in airway allergic inflammation, as revealed by increased total cell counts of inflammatory cells in BAL fluid, constituted mainly of eosinophils when compared to control group (Figures 2(b)-2(c)). We also found in TT/alum group an increased level of type 2 cytokines (IL-5 and IL-13) in BAL when compared to control group (Figures 2(d)-2(e)) thus, confirming the development of airway allergic inflammation. Having established in the OVA model the role of MyD88 and TRIF signaling in the prevention of allergic responses, we now tested the effects of absorbing two different preparations of LPS/lipid A onto tetanus toxoid/alum. One preparation was obtained from Escherichia coli 055:B5 that signals thorough TLR4 via MyD88 and TRIF pathways and other designated MPLA, which is a TRIF-biased TLR4 agonist [21]. As shown in Figure 2, the addition of LPS to tetanus toxoid alum preparation inhibited significantly the development allergic airway inflammation, as evidenced by lower number of total cell counts and eosinophils in BAL compared to TT/alum group (Figures 2(b)-2(c)). Also IL-5, but not IL-13, levels in BAL were significantly decreased in LPS group (Figures 2(d)-2(e)). In contrast, although Th2 responses of MPLA group were lower than TT/alum group, these responses did not reach statistical significance (Figure 2). We conclude tetanus toxoid adsorbed to alum behaves like an allergen and that LPS, but not MPLA, efficiently dampens alum pro-Th2 activity. To confirm this, we determined systemic antibody production by measuring serum levels of total IgE. As shown in Figure 3, sensitization and challenge with tetanus toxoid increased IgE levels when compared to control group. The addition of LPS, but not MPLA, to alum decreased significantly IgE levels (Figure 3(a)). Conversely, IgG1 antibodies against tetanus toxoid were similar in PBS, LPS, or MPLA groups (Figure 3(b)) while IgG2a specific antibodies were increased in LPS when compared to PBS group (Figure 3(c)). These results indicate that LPS prevented the production of IgE anaphylactic antibodies, did not interfere significantly in TT-specific IgG1, and augmented TT-specific IgG2a antibody production.


Adsorption of Toll-Like Receptor 4 Agonist to Alum-Based Tetanus Toxoid Vaccine Dampens Pro-T Helper 2 Activities and Enhances Antibody Responses.

Bortolatto J, Mirotti L, Rodriguez D, Gomes E, Russo M - J Immunol Res (2015)

Effects of adsorption of LPS or MPLA onto TT/alum sensitization on TT-mediated airway allergic responses. (a) Protocol: BALB/c mice sensitized with s.c. TT/alum in the presence or not of LPS (10 μg) or MPLA (10 μg) on days 0 and 7, and challenged with TT i.n. on days 14 and 21. Samples obtained on day 22: (b) total number of cells and (c) eosinophils in BAL; ((d) and (e)) levels of IL-5 and IL-13 in BAL. Data shown as mean ± SE, one-way ANOVA: #P < 0.05; different from TT/alum/PBS group (n = 5), and experiment was repeated twice.
© Copyright Policy - open-access
Related In: Results  -  Collection

Show All Figures
getmorefigures.php?uid=PMC4562177&req=5

fig2: Effects of adsorption of LPS or MPLA onto TT/alum sensitization on TT-mediated airway allergic responses. (a) Protocol: BALB/c mice sensitized with s.c. TT/alum in the presence or not of LPS (10 μg) or MPLA (10 μg) on days 0 and 7, and challenged with TT i.n. on days 14 and 21. Samples obtained on day 22: (b) total number of cells and (c) eosinophils in BAL; ((d) and (e)) levels of IL-5 and IL-13 in BAL. Data shown as mean ± SE, one-way ANOVA: #P < 0.05; different from TT/alum/PBS group (n = 5), and experiment was repeated twice.
Mentions: We next adapted the OVA model protocol to tetanus toxoid antigen. As depicted in Figure 2(a), sensitizations of tetanus toxoid adsorbed to alum (TT/alum group) followed by i.n. challenges resulted in airway allergic inflammation, as revealed by increased total cell counts of inflammatory cells in BAL fluid, constituted mainly of eosinophils when compared to control group (Figures 2(b)-2(c)). We also found in TT/alum group an increased level of type 2 cytokines (IL-5 and IL-13) in BAL when compared to control group (Figures 2(d)-2(e)) thus, confirming the development of airway allergic inflammation. Having established in the OVA model the role of MyD88 and TRIF signaling in the prevention of allergic responses, we now tested the effects of absorbing two different preparations of LPS/lipid A onto tetanus toxoid/alum. One preparation was obtained from Escherichia coli 055:B5 that signals thorough TLR4 via MyD88 and TRIF pathways and other designated MPLA, which is a TRIF-biased TLR4 agonist [21]. As shown in Figure 2, the addition of LPS to tetanus toxoid alum preparation inhibited significantly the development allergic airway inflammation, as evidenced by lower number of total cell counts and eosinophils in BAL compared to TT/alum group (Figures 2(b)-2(c)). Also IL-5, but not IL-13, levels in BAL were significantly decreased in LPS group (Figures 2(d)-2(e)). In contrast, although Th2 responses of MPLA group were lower than TT/alum group, these responses did not reach statistical significance (Figure 2). We conclude tetanus toxoid adsorbed to alum behaves like an allergen and that LPS, but not MPLA, efficiently dampens alum pro-Th2 activity. To confirm this, we determined systemic antibody production by measuring serum levels of total IgE. As shown in Figure 3, sensitization and challenge with tetanus toxoid increased IgE levels when compared to control group. The addition of LPS, but not MPLA, to alum decreased significantly IgE levels (Figure 3(a)). Conversely, IgG1 antibodies against tetanus toxoid were similar in PBS, LPS, or MPLA groups (Figure 3(b)) while IgG2a specific antibodies were increased in LPS when compared to PBS group (Figure 3(c)). These results indicate that LPS prevented the production of IgE anaphylactic antibodies, did not interfere significantly in TT-specific IgG1, and augmented TT-specific IgG2a antibody production.

Bottom Line: Here we found that alum-based tetanus toxoid (TT) vaccine was biased toward a Th-2 profile as shown by TT-induced airway eosinophilic inflammation, type 2 cytokine production, and high levels of IgE anaphylactic antibodies.Importantly, in a situation with antigenic competition (OVA plus TT), TT-specific IgG1 or IgG2a was decreased compared with TT sensitization.In conclusion, the addition of LPS induces a more robust IgG1 and IgG2a TT-specific antibody production and concomitantly decreases Th2-cellular and humoral responses, indicating a potential use of alum/TLR-based vaccines.

View Article: PubMed Central - PubMed

Affiliation: Department of Immunology, Institute of Biomedical Science, University of São Paulo, 05508-000 São Paulo, SP, Brazil.

ABSTRACT
Aluminum salts gels (alum) are TLR-independent adjuvants and have been used to boost antibody responses in alum-based vaccines such as diphtheria, pertussis, and tetanus toxoid (DPT) triple vaccine. However, the pro-Th2 activity of alum-based vaccine formulations has not been fully appreciated. Here we found that alum-based tetanus toxoid (TT) vaccine was biased toward a Th-2 profile as shown by TT-induced airway eosinophilic inflammation, type 2 cytokine production, and high levels of IgE anaphylactic antibodies. The adsorption into alum of prototypic TLR4 agonists such as lipopolysaccharides (LPS) derived from Escherichia coli consistently dampened TT-induced Th2 activities without inducing IFNγ or Th1-like responses in the lung. Conversely, adsorption of monophosphoryl lipid A (MPLA) extracted from Salmonella minnesota, which is a TIR-domain-containing adapter-inducing interferon-β- (TRIF-) biased TLR4 agonist, was less effective in decreasing Th-2 responses. Importantly, in a situation with antigenic competition (OVA plus TT), TT-specific IgG1 or IgG2a was decreased compared with TT sensitization. Notably, LPS increased the production of IgG1 and IgG2a TT-specific antibodies. In conclusion, the addition of LPS induces a more robust IgG1 and IgG2a TT-specific antibody production and concomitantly decreases Th2-cellular and humoral responses, indicating a potential use of alum/TLR-based vaccines.

No MeSH data available.


Related in: MedlinePlus