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Gpr97 Is Dispensable for Inflammation in OVA-Induced Asthmatic Mice.

Shi JP, Li XN, Zhang XY, Du B, Jiang WZ, Liu MY, Wang JJ, Wang ZG, Ren H, Qian M - PLoS ONE (2015)

Bottom Line: In our study, no obvious alteration in the inflammatory response or airway remodeling was found in the Gpr97-deficient mice with OVA-induced asthma.Neither the secretion of cytokines, including IL-4, IL-6 and IFN-γ, nor inflammatory cell recruitment was altered in the Gpr97-deficient mice.Our findings imply that Gpr97 might not be required for the development of airway inflammation in OVA-induced allergic asthma in mice.

View Article: PubMed Central - PubMed

Affiliation: Shanghai Key Laboratory of Regulatory Biology, School of Life Sciences, East China Normal University, Shanghai, China; Institute of Biomedical Sciences, School of Life Sciences, East China Normal University, Shanghai, China.

ABSTRACT

Background: Asthma is a complex inflammatory disorder involving the activation and invasion of various immune cells. GPR97 is highly expressed in some immunocytes, including mast cells and eosinophils, which play critical roles in asthma development. However, the role of Gpr97 in regulating airway inflammation in asthma has rarely been reported. In this study, we investigated the potential role of Gpr97 in the development of allergic asthma in mice.

Methods: Relevant airway asthmatic mouse models were constructed with both wild-type and Gpr97-/- mice sensitized to 250 μg ovalbumin (OVA). The levels of interleukin IL-4, IL-6 and IFN-γ, which are involved in OVA-induced asthma, in the bronchoalveolar lavage fluid (BALF) and the IgE level in the serum were examined by enzyme-linked immunosorbent assay (ELISA). The invasion of mast cells and eosinophils into lung tissues was assessed by immunohistochemical and eosinophil peroxidase activity assays, respectively. Goblet cell hyperplasia and mucus production were morphologically evaluated with periodic acid-Schiff (PAS) staining.

Results: In our study, no obvious alteration in the inflammatory response or airway remodeling was found in the Gpr97-deficient mice with OVA-induced asthma. Neither the secretion of cytokines, including IL-4, IL-6 and IFN-γ, nor inflammatory cell recruitment was altered in the Gpr97-deficient mice. Moreover, Gpr97 deficiency did not affect airway remodeling or mucus production in the asthma mouse model.

Conclusion: Our findings imply that Gpr97 might not be required for the development of airway inflammation in OVA-induced allergic asthma in mice.

No MeSH data available.


Related in: MedlinePlus

Detection of airway inflammation and morphological alterations following treatments with different concentrations of OVA in OVA-induced asthma mouse model.(A) ELISA analyses of the expression levels of IgE in the serum and IL-4, IL-6, and IFN-γ in the BALF following treatments with different concentrations of OVA in OVA-induced asthmatic mice. (B) The number of cells was quantified using a blood cell counting plate in the BALF for the different concentrations of OVA in OVA-induced asthma mice. (C) Eosinophil peroxidase activity was measured in the lung tissues of the asthmatic mice. (D) Lung edema was determined according to the lung/body weight data in OVA-induced asthma mice. (E) The staining of tissue for the detection of mast cells with anti-CD117 antibody in the lung tissues of the mice challenged with OVA. H&E staining of lung sections revealed the thickness of the alveolar walls, and PAS staining showed goblet cell hyperplasia along with mucus production in the lung tissues of the OVA-induced asthmatic mice. Lung sections were examined at the original magnification of 10× and at a higher magnification of 40×, as shown in the black square in the upper right corner of each picture. Scale bars: 100 μm in the panel. The data were normalized to the PBS sample data and are presented as the mean ± SEM (n = 6 animals for each genotype, as indicated by asterisks (* P < 0.05, ** P < 0.01, and *** P < 0.001).
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pone.0131461.g001: Detection of airway inflammation and morphological alterations following treatments with different concentrations of OVA in OVA-induced asthma mouse model.(A) ELISA analyses of the expression levels of IgE in the serum and IL-4, IL-6, and IFN-γ in the BALF following treatments with different concentrations of OVA in OVA-induced asthmatic mice. (B) The number of cells was quantified using a blood cell counting plate in the BALF for the different concentrations of OVA in OVA-induced asthma mice. (C) Eosinophil peroxidase activity was measured in the lung tissues of the asthmatic mice. (D) Lung edema was determined according to the lung/body weight data in OVA-induced asthma mice. (E) The staining of tissue for the detection of mast cells with anti-CD117 antibody in the lung tissues of the mice challenged with OVA. H&E staining of lung sections revealed the thickness of the alveolar walls, and PAS staining showed goblet cell hyperplasia along with mucus production in the lung tissues of the OVA-induced asthmatic mice. Lung sections were examined at the original magnification of 10× and at a higher magnification of 40×, as shown in the black square in the upper right corner of each picture. Scale bars: 100 μm in the panel. The data were normalized to the PBS sample data and are presented as the mean ± SEM (n = 6 animals for each genotype, as indicated by asterisks (* P < 0.05, ** P < 0.01, and *** P < 0.001).

Mentions: To study inflammation in OVA-induced allergic asthma mice, the expression levels of IL-4, IL-6, IFN-γ in the BALF and IgE in the serum were determined by ELISA [18–22]. We found that there were increases in the levels of IgE, IL-4, and IL-6 in an OVA concentration-dependent manner, while the level of IFN-γ, which is a Th1-related cytokine, was decreased (Fig 1A). To investigate inflammatory cell invasion, we counted the number of BALF cells. The total number of infiltrating cells in the BALF was increased under the OVA challenge (Fig 1B). At the same time, eosinophil invasion, which is characteristic of allergic asthma, was determined according to eosinophil peroxidase (EPO) activity. The results showed that there was a marked increase in eosinophil activity, even at the low concentration of OVA (Fig 1C). To explore changes in the lung airway wall in OVA-induced allergic asthma mice, the lungs were weighed, and the morphological characteristics were observed. The results showed that the lungs of the OVA-induced mice increased in weight (Fig 1D). On the other hand, an increase in CD117-positive mast cells was detected in the investigation of mast cell invasion in the lung tissues, which is anther characteristic of asthma. Immunohistochemical analysis of the slices showed that the amount of CD117-positive mast cells in the mouse lungs was much higher following induction with OVA (Fig 1E). The number of PAS-stained cells in the bronchus of the lung was also increased sharply, and airway epithelial extensions along with mucus production were evident under a light microscope (Fig 1E). According to our results, 250 μg ovalbumin, which was the suitable concentration for the sensitized treatment in the allergic asthma mouse model, was used for the Gpr97-deficient asthma mouse model.


Gpr97 Is Dispensable for Inflammation in OVA-Induced Asthmatic Mice.

Shi JP, Li XN, Zhang XY, Du B, Jiang WZ, Liu MY, Wang JJ, Wang ZG, Ren H, Qian M - PLoS ONE (2015)

Detection of airway inflammation and morphological alterations following treatments with different concentrations of OVA in OVA-induced asthma mouse model.(A) ELISA analyses of the expression levels of IgE in the serum and IL-4, IL-6, and IFN-γ in the BALF following treatments with different concentrations of OVA in OVA-induced asthmatic mice. (B) The number of cells was quantified using a blood cell counting plate in the BALF for the different concentrations of OVA in OVA-induced asthma mice. (C) Eosinophil peroxidase activity was measured in the lung tissues of the asthmatic mice. (D) Lung edema was determined according to the lung/body weight data in OVA-induced asthma mice. (E) The staining of tissue for the detection of mast cells with anti-CD117 antibody in the lung tissues of the mice challenged with OVA. H&E staining of lung sections revealed the thickness of the alveolar walls, and PAS staining showed goblet cell hyperplasia along with mucus production in the lung tissues of the OVA-induced asthmatic mice. Lung sections were examined at the original magnification of 10× and at a higher magnification of 40×, as shown in the black square in the upper right corner of each picture. Scale bars: 100 μm in the panel. The data were normalized to the PBS sample data and are presented as the mean ± SEM (n = 6 animals for each genotype, as indicated by asterisks (* P < 0.05, ** P < 0.01, and *** P < 0.001).
© Copyright Policy
Related In: Results  -  Collection

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getmorefigures.php?uid=PMC4489018&req=5

pone.0131461.g001: Detection of airway inflammation and morphological alterations following treatments with different concentrations of OVA in OVA-induced asthma mouse model.(A) ELISA analyses of the expression levels of IgE in the serum and IL-4, IL-6, and IFN-γ in the BALF following treatments with different concentrations of OVA in OVA-induced asthmatic mice. (B) The number of cells was quantified using a blood cell counting plate in the BALF for the different concentrations of OVA in OVA-induced asthma mice. (C) Eosinophil peroxidase activity was measured in the lung tissues of the asthmatic mice. (D) Lung edema was determined according to the lung/body weight data in OVA-induced asthma mice. (E) The staining of tissue for the detection of mast cells with anti-CD117 antibody in the lung tissues of the mice challenged with OVA. H&E staining of lung sections revealed the thickness of the alveolar walls, and PAS staining showed goblet cell hyperplasia along with mucus production in the lung tissues of the OVA-induced asthmatic mice. Lung sections were examined at the original magnification of 10× and at a higher magnification of 40×, as shown in the black square in the upper right corner of each picture. Scale bars: 100 μm in the panel. The data were normalized to the PBS sample data and are presented as the mean ± SEM (n = 6 animals for each genotype, as indicated by asterisks (* P < 0.05, ** P < 0.01, and *** P < 0.001).
Mentions: To study inflammation in OVA-induced allergic asthma mice, the expression levels of IL-4, IL-6, IFN-γ in the BALF and IgE in the serum were determined by ELISA [18–22]. We found that there were increases in the levels of IgE, IL-4, and IL-6 in an OVA concentration-dependent manner, while the level of IFN-γ, which is a Th1-related cytokine, was decreased (Fig 1A). To investigate inflammatory cell invasion, we counted the number of BALF cells. The total number of infiltrating cells in the BALF was increased under the OVA challenge (Fig 1B). At the same time, eosinophil invasion, which is characteristic of allergic asthma, was determined according to eosinophil peroxidase (EPO) activity. The results showed that there was a marked increase in eosinophil activity, even at the low concentration of OVA (Fig 1C). To explore changes in the lung airway wall in OVA-induced allergic asthma mice, the lungs were weighed, and the morphological characteristics were observed. The results showed that the lungs of the OVA-induced mice increased in weight (Fig 1D). On the other hand, an increase in CD117-positive mast cells was detected in the investigation of mast cell invasion in the lung tissues, which is anther characteristic of asthma. Immunohistochemical analysis of the slices showed that the amount of CD117-positive mast cells in the mouse lungs was much higher following induction with OVA (Fig 1E). The number of PAS-stained cells in the bronchus of the lung was also increased sharply, and airway epithelial extensions along with mucus production were evident under a light microscope (Fig 1E). According to our results, 250 μg ovalbumin, which was the suitable concentration for the sensitized treatment in the allergic asthma mouse model, was used for the Gpr97-deficient asthma mouse model.

Bottom Line: In our study, no obvious alteration in the inflammatory response or airway remodeling was found in the Gpr97-deficient mice with OVA-induced asthma.Neither the secretion of cytokines, including IL-4, IL-6 and IFN-γ, nor inflammatory cell recruitment was altered in the Gpr97-deficient mice.Our findings imply that Gpr97 might not be required for the development of airway inflammation in OVA-induced allergic asthma in mice.

View Article: PubMed Central - PubMed

Affiliation: Shanghai Key Laboratory of Regulatory Biology, School of Life Sciences, East China Normal University, Shanghai, China; Institute of Biomedical Sciences, School of Life Sciences, East China Normal University, Shanghai, China.

ABSTRACT

Background: Asthma is a complex inflammatory disorder involving the activation and invasion of various immune cells. GPR97 is highly expressed in some immunocytes, including mast cells and eosinophils, which play critical roles in asthma development. However, the role of Gpr97 in regulating airway inflammation in asthma has rarely been reported. In this study, we investigated the potential role of Gpr97 in the development of allergic asthma in mice.

Methods: Relevant airway asthmatic mouse models were constructed with both wild-type and Gpr97-/- mice sensitized to 250 μg ovalbumin (OVA). The levels of interleukin IL-4, IL-6 and IFN-γ, which are involved in OVA-induced asthma, in the bronchoalveolar lavage fluid (BALF) and the IgE level in the serum were examined by enzyme-linked immunosorbent assay (ELISA). The invasion of mast cells and eosinophils into lung tissues was assessed by immunohistochemical and eosinophil peroxidase activity assays, respectively. Goblet cell hyperplasia and mucus production were morphologically evaluated with periodic acid-Schiff (PAS) staining.

Results: In our study, no obvious alteration in the inflammatory response or airway remodeling was found in the Gpr97-deficient mice with OVA-induced asthma. Neither the secretion of cytokines, including IL-4, IL-6 and IFN-γ, nor inflammatory cell recruitment was altered in the Gpr97-deficient mice. Moreover, Gpr97 deficiency did not affect airway remodeling or mucus production in the asthma mouse model.

Conclusion: Our findings imply that Gpr97 might not be required for the development of airway inflammation in OVA-induced allergic asthma in mice.

No MeSH data available.


Related in: MedlinePlus