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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 PIC (TLR3) or LPS (TLR4) agonists onto OVA/alum sensitization on OVA-induced cellular and humoral responses. (a) Protocol: C57BL/6 WT mice sensitized with s.c. OVA/alum in the presence or not of PIC (10 μg) or LPS (10 μg) on days 0 and 7 and challenged with OVA i.n. on days 14 and 21. Samples obtained on day 22. (b) Total number of cells and (c) eosinophils in BAL; ((d), (e), and (f)) levels of IL-5, IL-13, and IFN- g in BAL. ((g), (h), and (i)) Levels of serum isotypes: total IgE, OVA-specific IgG1, and OVA-specific IgG2a. Data shown as mean ± SE, one-way ANOVA: #P < 0.05 different from OVA/alum/PBS group (n = 5), and experiment was repeated twice.
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fig1: Effects of adsorption of PIC (TLR3) or LPS (TLR4) agonists onto OVA/alum sensitization on OVA-induced cellular and humoral responses. (a) Protocol: C57BL/6 WT mice sensitized with s.c. OVA/alum in the presence or not of PIC (10 μg) or LPS (10 μg) on days 0 and 7 and challenged with OVA i.n. on days 14 and 21. Samples obtained on day 22. (b) Total number of cells and (c) eosinophils in BAL; ((d), (e), and (f)) levels of IL-5, IL-13, and IFN- g in BAL. ((g), (h), and (i)) Levels of serum isotypes: total IgE, OVA-specific IgG1, and OVA-specific IgG2a. Data shown as mean ± SE, one-way ANOVA: #P < 0.05 different from OVA/alum/PBS group (n = 5), and experiment was repeated twice.

Mentions: We have previously shown that TLR4 agonist (LPS) adsorbed to OVA/alum prevented the development of asthma-like responses via MyD88, but not TRIF pathway [18]. In order to ascertain more directly the effect of TRIF signaling, we used the OVA model to compare the effect of Poly I:C, a TLR3 synthetic agonist analog of dsRNA, which signals solely thorough TRIF; with LPS, a TLR4 agonist that signals thorough MyD88 and TRIF pathways. For this, BALB/c mice were sensitized to OVA adsorbed to alum in the absence (allergic group) or presence of agonists of TLR3 (Poly-I:C) or TLR4 (LPS). Overall, both TLRs agonists when adsorbed to OVA/alum dampened Th2 responses when compared to allergic (OVA/alum) group (Figure 1). However, LPS was consistently more effective than PIC in decreasing total cell counts and eosinophil number in BAL fluid (Figures 1(b)-1(c)), IL-5, and IL-13 production (Figures 1(d)-1(e)), and IgE levels (Figure 1(g)). Importantly, the levels of IFNγ in BAL in PIC or LPS groups were not increased and were similar to naive or allergic (PBS) groups (Figure 1(f)). Regarding antibody production, again LPS was more effective than PIC in decreasing IgE (Figure 1(g)). PIC but not LPS decreased OVA-specific IgG1 isotype (Figure 1(h)) while LPS increased IgG2a production (Figure 1(i)). Altogether, these results indicate that LPS was more efficient than PIC in inhibiting Th2-mediated airway allergic response.


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 PIC (TLR3) or LPS (TLR4) agonists onto OVA/alum sensitization on OVA-induced cellular and humoral responses. (a) Protocol: C57BL/6 WT mice sensitized with s.c. OVA/alum in the presence or not of PIC (10 μg) or LPS (10 μg) on days 0 and 7 and challenged with OVA i.n. on days 14 and 21. Samples obtained on day 22. (b) Total number of cells and (c) eosinophils in BAL; ((d), (e), and (f)) levels of IL-5, IL-13, and IFN- g in BAL. ((g), (h), and (i)) Levels of serum isotypes: total IgE, OVA-specific IgG1, and OVA-specific IgG2a. Data shown as mean ± SE, one-way ANOVA: #P < 0.05 different from OVA/alum/PBS group (n = 5), and experiment was repeated twice.
© Copyright Policy - open-access
Related In: Results  -  Collection

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getmorefigures.php?uid=PMC4562177&req=5

fig1: Effects of adsorption of PIC (TLR3) or LPS (TLR4) agonists onto OVA/alum sensitization on OVA-induced cellular and humoral responses. (a) Protocol: C57BL/6 WT mice sensitized with s.c. OVA/alum in the presence or not of PIC (10 μg) or LPS (10 μg) on days 0 and 7 and challenged with OVA i.n. on days 14 and 21. Samples obtained on day 22. (b) Total number of cells and (c) eosinophils in BAL; ((d), (e), and (f)) levels of IL-5, IL-13, and IFN- g in BAL. ((g), (h), and (i)) Levels of serum isotypes: total IgE, OVA-specific IgG1, and OVA-specific IgG2a. Data shown as mean ± SE, one-way ANOVA: #P < 0.05 different from OVA/alum/PBS group (n = 5), and experiment was repeated twice.
Mentions: We have previously shown that TLR4 agonist (LPS) adsorbed to OVA/alum prevented the development of asthma-like responses via MyD88, but not TRIF pathway [18]. In order to ascertain more directly the effect of TRIF signaling, we used the OVA model to compare the effect of Poly I:C, a TLR3 synthetic agonist analog of dsRNA, which signals solely thorough TRIF; with LPS, a TLR4 agonist that signals thorough MyD88 and TRIF pathways. For this, BALB/c mice were sensitized to OVA adsorbed to alum in the absence (allergic group) or presence of agonists of TLR3 (Poly-I:C) or TLR4 (LPS). Overall, both TLRs agonists when adsorbed to OVA/alum dampened Th2 responses when compared to allergic (OVA/alum) group (Figure 1). However, LPS was consistently more effective than PIC in decreasing total cell counts and eosinophil number in BAL fluid (Figures 1(b)-1(c)), IL-5, and IL-13 production (Figures 1(d)-1(e)), and IgE levels (Figure 1(g)). Importantly, the levels of IFNγ in BAL in PIC or LPS groups were not increased and were similar to naive or allergic (PBS) groups (Figure 1(f)). Regarding antibody production, again LPS was more effective than PIC in decreasing IgE (Figure 1(g)). PIC but not LPS decreased OVA-specific IgG1 isotype (Figure 1(h)) while LPS increased IgG2a production (Figure 1(i)). Altogether, these results indicate that LPS was more efficient than PIC in inhibiting Th2-mediated airway allergic response.

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