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Objective: In allergic asthma, regulatory T cell (Treg) number and function are decreased. Antigen-primed CD8+ T cells play an indispensable role in the full development of airway inflammation and airway hyper-responsiveness (AHR) occurring in asthma. In this study, we investigated the relationship between subpopulations of CD8+ T cells and CD39+ Tregs. Methods: Female C57BL/6 mice were used to develop the model of allergic asthma. Experimental mice were immunized with ovalbumin (OVA) by intra-peritoneal (i.p) injection and then challenged with OVA by intra-tracheal administration. Control mice were immunized with vehicle by i.p injection and challenged with OVA. Airway inflammation was determined by histology and AHR was measured by an invasive method. Levels of interferon (IFN)-γ, IL-4, and IL-17 in bronchoalveolar lavage fluid (BALF) were determined by enzyme-linked immunosorbent assay. The frequencies of CD8+IFN-γ+ cells (Tc1), CD8+IL-4+ cells (Tc2), CD8+IL-17+cells (Tc17), and CD39+Tregs were measured by flow cytometry. The correlation between CD39+Tregs and Tc subsets was analyzed by Pearson's test. Results: Experimental mice displayed phenotypes of allergic asthma, including inflammatory cell infiltration into the lungs, goblet cell hyperplasia, increased airway resistance, and increased IL-4 and IL-17 in BALF. Compared to control mice, experimental mice displayed lower CD39+Tregs and Tc1 but higher Tc2 and Tc17. There was a negative correlation between CD39+Tregs and Tc2 or Tc17. Conclusion: In allergic asthma, increased Tc2 and Tc17 are possibly related to insufficient CD39+Tregs. [ABSTRACT FROM AUTHOR]

Objective: In allergic asthma, regulatory T cell (Treg) number and function are decreased. Antigen-primed CD8 + T cells play an indispensable role in the full development of airway inflammation and airway hyper-responsiveness (AHR) occurring in asthma. In this study, we investigated the relationship between subpopulations of CD8 + T cells and CD39 + Tregs., Methods: Female C57BL/6 mice were used to develop the model of allergic asthma. Experimental mice were immunized with ovalbumin (OVA) by intra-peritoneal (i.p) injection and then challenged with OVA by intra-tracheal administration. Control mice were immunized with vehicle by i.p injection and challenged with OVA. Airway inflammation was determined by histology and AHR was measured by an invasive method. Levels of interferon (IFN)-γ, IL-4, and IL-17 in bronchoalveolar lavage fluid (BALF) were determined by enzyme-linked immunosorbent assay. The frequencies of CD8 + IFN-γ + cells (Tc1), CD8 + IL-4 + cells (Tc2), CD8 + IL-17 + cells (Tc17), and CD39 + Tregs were measured by flow cytometry. The correlation between CD39 + Tregs and Tc subsets was analyzed by Pearson's test., Results: Experimental mice displayed phenotypes of allergic asthma, including inflammatory cell infiltration into the lungs, goblet cell hyperplasia, increased airway resistance, and increased IL-4 and IL-17 in BALF. Compared to control mice, experimental mice displayed lower CD39 + Tregs and Tc1 but higher Tc2 and Tc17. There was a negative correlation between CD39 + Tregs and Tc2 or Tc17., Conclusion: In allergic asthma, increased Tc2 and Tc17 are possibly related to insufficient CD39 + Tregs.

Aim: To create a method of detecting typeII natural killer T (NKT) cells of mice., Methods: Biotinylated mouse CD1d monomers were mixed with sulfatide at a molar ratio of 1:3 (protein:lipid) and incubated at room temperature overnight, and then 80 μg of streptavidin-PE was added into 200 μg of the CD1d-sulfatide mixture and incubated at room temperature for 4 h to get sulfatide/CD1d tetramer. Flow cytometry was used to detect the percentage of typeII NKT cells in mononuclear cells (MNCs) of lung and spleen of normal mice, as well as the percentage of typeII NKT cells in spleen MNCs of mice after stimulated with sulfatide., Results: In normal mice, the percentage of typeII NKT cells accounted for (0.875±0.096)% and (1.175±0.263)% in MNCs of spleen and lung; the percentage in spleen MNCs after activated with sulfatide was (2.75±0.603)%, which significantly increased as compared with that in normal mice (P<0.01)., Conclusion: Sulfatide-loaded CD1d tetramer is an effective method of detecting typeII NKT cells in mice.