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Synthesized OVA323-339MAP octamers mitigate OVA-induced airway inflammation by regulating Foxp3 T regulatory cells.

Su W, Zhong W, Zhang Y, Xia Z - BMC Immunol. (2012)

Bottom Line: These results were further correlated with the decreased OVA specific immunoglobulin E (sIgE) level and the infiltration of inflammatory cells such as eosinophils and lymphocytes in BALF.However, OVA323-339 peptide monomers did not show any of the mentioned effects in the same animal model.Our study indicates that OVA323-339MAP had significant therapeutic effects on mice allergic airway inflammation by regulating the balance of Th1/Th2 response through Treg cells in vivo.

View Article: PubMed Central - HTML - PubMed

Affiliation: Department of Pediatrics, Ruijin Hospital Affiliated to Shanghai Jiao Tong University School of Medicine, Shanghai, China.

ABSTRACT

Background: Antigen-specific immunotherapy (SIT) has been widely practiced in treating allergic diseases such as asthma. However, this therapy may induce a series of allergic adverse events during treatment. Peptide immunotherapy (PIT) was explored to overcome these disadvantages. We confirmed that multiple antigen peptides (MAPs) do not cause autoimmune responses, which led to the presumption that MAPs intervention could alleviate allergic airway inflammation without inducing adverse effects.

Results: In this study, synthesized OVA323-339MAP octamers were subcutaneously injected into ovalbumin (OVA)-sensitized and -challenged Balb/c mice to observe its effect on allergic airway inflammation, Th2 immune response, and immune regulating function. It was confirmed that OVA sensitization and challenge led to significant peritracheal inflammatory, cell infiltration, and intensive Th2 response. Treatment of OVA323-339MAP octomers in the airway inflammation mice model increased CD4+CD25+Foxp3+ T regulatory (Treg) cells and their regulatory function in peripheral blood, mediastinal draining lymph nodes, and the spleen. Furthermore, OVA323-339MAP increased IL-10 levels in bronchial alveolar lavage fluid (BALF); up-regulated the expression of IL-10, membrane-bound TGF-β1, as well as Foxp3 in lung tissues; and up-regulated programmed death-1 (PD-1) and cytotoxic T lymphocyte associated antigen 4 (CTLA-4) on the surface of Treg cells. These results were further correlated with the decreased OVA specific immunoglobulin E (sIgE) level and the infiltration of inflammatory cells such as eosinophils and lymphocytes in BALF. However, OVA323-339 peptide monomers did not show any of the mentioned effects in the same animal model.

Conclusions: Our study indicates that OVA323-339MAP had significant therapeutic effects on mice allergic airway inflammation by regulating the balance of Th1/Th2 response through Treg cells in vivo.

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Related in: MedlinePlus

Th2 cells differentiation in spleen. Splenocytes were co-cultured with OVA (1 mg/ml) in vitro for the examination of Th2 cells differentiation. OVA323-339MAP intervention could significantly reduce the ratio of CD4+IL-4+ T cells compared with the OVA group (gating on CD4+ T cell populations). ELISA detection of the cytokines in supernatant showed that IL-4, IL-5, and IL-13 levels were significantly increased in the OVA group compared with the control group, but OVA323-339MAP intervention could significantly suppress their secretion and enhance the expression of IFN-γ, then the ratio of IFN-γ/IL-4 was increased. (* compared with the control group, P < 0.05; # compared with the OVA group, P < 0.05). Data represent one of three independent experiments.
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Figure 4: Th2 cells differentiation in spleen. Splenocytes were co-cultured with OVA (1 mg/ml) in vitro for the examination of Th2 cells differentiation. OVA323-339MAP intervention could significantly reduce the ratio of CD4+IL-4+ T cells compared with the OVA group (gating on CD4+ T cell populations). ELISA detection of the cytokines in supernatant showed that IL-4, IL-5, and IL-13 levels were significantly increased in the OVA group compared with the control group, but OVA323-339MAP intervention could significantly suppress their secretion and enhance the expression of IFN-γ, then the ratio of IFN-γ/IL-4 was increased. (* compared with the control group, P < 0.05; # compared with the OVA group, P < 0.05). Data represent one of three independent experiments.

Mentions: It is well documented that OVA-induced airway inflammation in Balb/c mice is mainly mediated by a Th2 response. In light of the finding that OVA323-339MAP can attenuate allergic airway inflammation, we asked if OVA323-339MAP treatment alters the OVA-induced Th2 response in the host. To this end, we measured the expression of Th1/Th2-related cytokines (IFN-γ, IL-4, IL-5 and IL-13) and the ratio of IFN-γ/IL-4 in the BALF of these mice. The levels of IL-4, IL-5 and IL-13 were significantly elevated in mice challenged with OVA (Figure 3 A, D, E), while the ratio of IFN-γ/IL-4 decreased significantly (Figure 3 C). However, OVA323-339MAP intervention reduced local Th2 cytokines production and increased the level of IFN-γ and the ratio of IFN-γ/IL-4 (Figure 3 A ~ E). Furthermore, in order to determine if OVA323-339MAP treatment influences antigen recall response in mice sensitized with OVA, splenocytes from control and OVA-challenged mice with or without OVA323-339MAP intervention were harvested. The IL-4 and IFN-γ positive populations were analyzed by flow cytometry (FCM) in CD4+ T cells and the amount of IL-4, IL-5, IL-13 and IFN-γ in the cell culture media were quantified by ELISA. When the lymphocytes were re-stimulated with OVA in vitro, CD4+ T cells from OVA-challenged mice exhibited a significantly higher IL-4 production than the control group. In vivo treatment with OVA323-339MAP virtually blocked Th2 cytokines production in response to OVA re-stimulation (Figure 4 A, G). Likewise, ELISA demonstrated that splenocytes from OVA-challenged mice exhibited a significantly higher IL-4, IL-5 and IL-13 response to antigen re-stimulation, while this antigen recall response was diminished in OVA323-339MAP-treated mice (Figure 4 B, E, F). Interestingly, OVA323-339MAP intervention caused more IFN-γ products while the levels of IFN-γ between the control and airway inflammation groups were biologically insignificant (Figure 4 C). These results suggest that OVA323-339MAP may alter Th2 response in this allergic airway model.


Synthesized OVA323-339MAP octamers mitigate OVA-induced airway inflammation by regulating Foxp3 T regulatory cells.

Su W, Zhong W, Zhang Y, Xia Z - BMC Immunol. (2012)

Th2 cells differentiation in spleen. Splenocytes were co-cultured with OVA (1 mg/ml) in vitro for the examination of Th2 cells differentiation. OVA323-339MAP intervention could significantly reduce the ratio of CD4+IL-4+ T cells compared with the OVA group (gating on CD4+ T cell populations). ELISA detection of the cytokines in supernatant showed that IL-4, IL-5, and IL-13 levels were significantly increased in the OVA group compared with the control group, but OVA323-339MAP intervention could significantly suppress their secretion and enhance the expression of IFN-γ, then the ratio of IFN-γ/IL-4 was increased. (* compared with the control group, P < 0.05; # compared with the OVA group, P < 0.05). Data represent one of three independent experiments.
© Copyright Policy - open-access
Related In: Results  -  Collection

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Figure 4: Th2 cells differentiation in spleen. Splenocytes were co-cultured with OVA (1 mg/ml) in vitro for the examination of Th2 cells differentiation. OVA323-339MAP intervention could significantly reduce the ratio of CD4+IL-4+ T cells compared with the OVA group (gating on CD4+ T cell populations). ELISA detection of the cytokines in supernatant showed that IL-4, IL-5, and IL-13 levels were significantly increased in the OVA group compared with the control group, but OVA323-339MAP intervention could significantly suppress their secretion and enhance the expression of IFN-γ, then the ratio of IFN-γ/IL-4 was increased. (* compared with the control group, P < 0.05; # compared with the OVA group, P < 0.05). Data represent one of three independent experiments.
Mentions: It is well documented that OVA-induced airway inflammation in Balb/c mice is mainly mediated by a Th2 response. In light of the finding that OVA323-339MAP can attenuate allergic airway inflammation, we asked if OVA323-339MAP treatment alters the OVA-induced Th2 response in the host. To this end, we measured the expression of Th1/Th2-related cytokines (IFN-γ, IL-4, IL-5 and IL-13) and the ratio of IFN-γ/IL-4 in the BALF of these mice. The levels of IL-4, IL-5 and IL-13 were significantly elevated in mice challenged with OVA (Figure 3 A, D, E), while the ratio of IFN-γ/IL-4 decreased significantly (Figure 3 C). However, OVA323-339MAP intervention reduced local Th2 cytokines production and increased the level of IFN-γ and the ratio of IFN-γ/IL-4 (Figure 3 A ~ E). Furthermore, in order to determine if OVA323-339MAP treatment influences antigen recall response in mice sensitized with OVA, splenocytes from control and OVA-challenged mice with or without OVA323-339MAP intervention were harvested. The IL-4 and IFN-γ positive populations were analyzed by flow cytometry (FCM) in CD4+ T cells and the amount of IL-4, IL-5, IL-13 and IFN-γ in the cell culture media were quantified by ELISA. When the lymphocytes were re-stimulated with OVA in vitro, CD4+ T cells from OVA-challenged mice exhibited a significantly higher IL-4 production than the control group. In vivo treatment with OVA323-339MAP virtually blocked Th2 cytokines production in response to OVA re-stimulation (Figure 4 A, G). Likewise, ELISA demonstrated that splenocytes from OVA-challenged mice exhibited a significantly higher IL-4, IL-5 and IL-13 response to antigen re-stimulation, while this antigen recall response was diminished in OVA323-339MAP-treated mice (Figure 4 B, E, F). Interestingly, OVA323-339MAP intervention caused more IFN-γ products while the levels of IFN-γ between the control and airway inflammation groups were biologically insignificant (Figure 4 C). These results suggest that OVA323-339MAP may alter Th2 response in this allergic airway model.

Bottom Line: These results were further correlated with the decreased OVA specific immunoglobulin E (sIgE) level and the infiltration of inflammatory cells such as eosinophils and lymphocytes in BALF.However, OVA323-339 peptide monomers did not show any of the mentioned effects in the same animal model.Our study indicates that OVA323-339MAP had significant therapeutic effects on mice allergic airway inflammation by regulating the balance of Th1/Th2 response through Treg cells in vivo.

View Article: PubMed Central - HTML - PubMed

Affiliation: Department of Pediatrics, Ruijin Hospital Affiliated to Shanghai Jiao Tong University School of Medicine, Shanghai, China.

ABSTRACT

Background: Antigen-specific immunotherapy (SIT) has been widely practiced in treating allergic diseases such as asthma. However, this therapy may induce a series of allergic adverse events during treatment. Peptide immunotherapy (PIT) was explored to overcome these disadvantages. We confirmed that multiple antigen peptides (MAPs) do not cause autoimmune responses, which led to the presumption that MAPs intervention could alleviate allergic airway inflammation without inducing adverse effects.

Results: In this study, synthesized OVA323-339MAP octamers were subcutaneously injected into ovalbumin (OVA)-sensitized and -challenged Balb/c mice to observe its effect on allergic airway inflammation, Th2 immune response, and immune regulating function. It was confirmed that OVA sensitization and challenge led to significant peritracheal inflammatory, cell infiltration, and intensive Th2 response. Treatment of OVA323-339MAP octomers in the airway inflammation mice model increased CD4+CD25+Foxp3+ T regulatory (Treg) cells and their regulatory function in peripheral blood, mediastinal draining lymph nodes, and the spleen. Furthermore, OVA323-339MAP increased IL-10 levels in bronchial alveolar lavage fluid (BALF); up-regulated the expression of IL-10, membrane-bound TGF-β1, as well as Foxp3 in lung tissues; and up-regulated programmed death-1 (PD-1) and cytotoxic T lymphocyte associated antigen 4 (CTLA-4) on the surface of Treg cells. These results were further correlated with the decreased OVA specific immunoglobulin E (sIgE) level and the infiltration of inflammatory cells such as eosinophils and lymphocytes in BALF. However, OVA323-339 peptide monomers did not show any of the mentioned effects in the same animal model.

Conclusions: Our study indicates that OVA323-339MAP had significant therapeutic effects on mice allergic airway inflammation by regulating the balance of Th1/Th2 response through Treg cells in vivo.

Show MeSH
Related in: MedlinePlus