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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 LPS onto OVA/TT/alum sensitization on OVA-induced airway allergic inflammation. (a) Protocol: BALB/c mice sensitized with s.c. OVA/TT/alum in the presence or not of LPS (10 μg) on days 0 and 7 and challenged with i.n. OVA 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 IFN-γ in BAL. Data shown as mean ± SE, one-way ANOVA: #P < 0.05; different from OVA/TT/alum/PBS group (n = 5), and experiment was repeated twice.
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fig4: Effects of adsorption of LPS onto OVA/TT/alum sensitization on OVA-induced airway allergic inflammation. (a) Protocol: BALB/c mice sensitized with s.c. OVA/TT/alum in the presence or not of LPS (10 μg) on days 0 and 7 and challenged with i.n. OVA 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 IFN-γ in BAL. Data shown as mean ± SE, one-way ANOVA: #P < 0.05; different from OVA/TT/alum/PBS group (n = 5), and experiment was repeated twice.

Mentions: The benefits of combined vaccines such as diphtheria, pertussis, tetanus (DPT) vaccine are multiple including fewer injections and lower cost. However, special attention must be taken with the immunogenicity of individual antigens when combining unrelated antigens since it can be impaired, enhanced, or not affected [23, 24]. To test antigen combination in our model, we used OVA and TT antigens as unrelated antigens, adsorbing them to alum in the presence or absence of LPS, as depicted in Figure 4(a). To monitor Th2 activities we determined OVA-specific responses such as airway allergic inflammation and serum titers of anaphylactic IgE after i.n. OVA challenge. TT-specific responses were determined by serum isotype antibody production after TT s.c challenge. Regarding OVA-specific airway allergic responses, we found that addition of LPS inhibited total cells and eosinophil numbers recruitment to BAL when compared to (PBS) allergic group (Figures 4(b)-4(c)). In addition, IL-5 and, of note, IFNγ levels in BAL were also decreased in LPS group as compared to PBS group (Figures 4(d)-4(e)), indicating that LPS do not shift towards an OVA-induced lung Th1 response. As expected, LPS blocked OVA-specific IgE anaphylactic antibodies as revealed by passive cutaneous anaphylaxis titers (Figure 5(a)), thus confirming the antiallergic effect of LPS in our model. In addition, OVA-specific IgG1 and IgG2a antibodies were increased in LPS group compared to PBS group (Figures 5(b)-5(c)). Regarding TT-specific responses, LPS addition showed a similar effect to that obtained with OVA-specific responses (Figures 5(d)–5(f)). Of note, we found that the concentrations of TT-specific antibodies (IgG1 and IgG2a) were lower (Figures 5(e)-5(f)) than those obtained when sensitization and challenge were done with TT alone (Figures 3(b)-3(c)), indicating that the phenomenon known as antigenic completion has occurred [24]. Nevertheless and, notably, addition of LPS to OVA/TT/alum resulted in a higher production of IgG1 and IgG2a anti-TT antibodies when compared to TT/OVA/alum group. As expected, TT-specific IgE titers were lower in LPS group when compared with PBS group (Figure 5(d)). Therefore, the addition of LPS to alum-based vaccine with two unrelated antigens was able to increase IgG1 and IgG2a antibody production and decrease anaphylactic IgE titers, a more desirable profile for neutralizing antibodies.


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 onto OVA/TT/alum sensitization on OVA-induced airway allergic inflammation. (a) Protocol: BALB/c mice sensitized with s.c. OVA/TT/alum in the presence or not of LPS (10 μg) on days 0 and 7 and challenged with i.n. OVA 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 IFN-γ in BAL. Data shown as mean ± SE, one-way ANOVA: #P < 0.05; different from OVA/TT/alum/PBS group (n = 5), and experiment was repeated twice.
© Copyright Policy - open-access
Related In: Results  -  Collection

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fig4: Effects of adsorption of LPS onto OVA/TT/alum sensitization on OVA-induced airway allergic inflammation. (a) Protocol: BALB/c mice sensitized with s.c. OVA/TT/alum in the presence or not of LPS (10 μg) on days 0 and 7 and challenged with i.n. OVA 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 IFN-γ in BAL. Data shown as mean ± SE, one-way ANOVA: #P < 0.05; different from OVA/TT/alum/PBS group (n = 5), and experiment was repeated twice.
Mentions: The benefits of combined vaccines such as diphtheria, pertussis, tetanus (DPT) vaccine are multiple including fewer injections and lower cost. However, special attention must be taken with the immunogenicity of individual antigens when combining unrelated antigens since it can be impaired, enhanced, or not affected [23, 24]. To test antigen combination in our model, we used OVA and TT antigens as unrelated antigens, adsorbing them to alum in the presence or absence of LPS, as depicted in Figure 4(a). To monitor Th2 activities we determined OVA-specific responses such as airway allergic inflammation and serum titers of anaphylactic IgE after i.n. OVA challenge. TT-specific responses were determined by serum isotype antibody production after TT s.c challenge. Regarding OVA-specific airway allergic responses, we found that addition of LPS inhibited total cells and eosinophil numbers recruitment to BAL when compared to (PBS) allergic group (Figures 4(b)-4(c)). In addition, IL-5 and, of note, IFNγ levels in BAL were also decreased in LPS group as compared to PBS group (Figures 4(d)-4(e)), indicating that LPS do not shift towards an OVA-induced lung Th1 response. As expected, LPS blocked OVA-specific IgE anaphylactic antibodies as revealed by passive cutaneous anaphylaxis titers (Figure 5(a)), thus confirming the antiallergic effect of LPS in our model. In addition, OVA-specific IgG1 and IgG2a antibodies were increased in LPS group compared to PBS group (Figures 5(b)-5(c)). Regarding TT-specific responses, LPS addition showed a similar effect to that obtained with OVA-specific responses (Figures 5(d)–5(f)). Of note, we found that the concentrations of TT-specific antibodies (IgG1 and IgG2a) were lower (Figures 5(e)-5(f)) than those obtained when sensitization and challenge were done with TT alone (Figures 3(b)-3(c)), indicating that the phenomenon known as antigenic completion has occurred [24]. Nevertheless and, notably, addition of LPS to OVA/TT/alum resulted in a higher production of IgG1 and IgG2a anti-TT antibodies when compared to TT/OVA/alum group. As expected, TT-specific IgE titers were lower in LPS group when compared with PBS group (Figure 5(d)). Therefore, the addition of LPS to alum-based vaccine with two unrelated antigens was able to increase IgG1 and IgG2a antibody production and decrease anaphylactic IgE titers, a more desirable profile for neutralizing antibodies.

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