Correlation of Local FOXP3-Expressing T Cells and Th1-Th2 Balance in Perennial Allergic Nasal Mucosa

Abstract

Regulatory T cells (Treg) play some important roles in allergic rhinitis. The most specific marker for Treg is FOXP3, a recently identified transcription factor that is essential for Treg development. In order to clarify the levels of Treg in allergic nasal mucosa, we studied the relationship between FOXP3-expressing cells and Th1-Th2 balance in nasal mucosa by means of immunohistochemistry. Human turbinates were obtained after turbinectomy from 26 patients (14 patients with perennial allergic rhinitis and 12 patients with nonallergic rhinitis). To identify the cells expressing the FOXP3 protein, double immunostaining was performed by using anti-FOXP3 antibody and anti-CD3 antibody. There was no significant difference in the percentage of FOXP3+CD3+ cells among CD3+ cells in the nasal mucosa of two groups. The proportion of FOXP3+CD3+ cells tend to be correlated positively with GATA3+CD3+ cells/T-bet+CD3+ cells ratio (R = 0.56, P = 0.04). A positive correlation with GATA3+CD3+/T-bet+CD3+ ratio and FOXP3+CD3+/CD3+ ratio suggests the role of local regulatory T cells as a minimal control of the chronic allergen exposure in nasal mucosa.

Figure 1

Identification of the cells expressing FOXP3 among CD3 positive cells in human allergic nasal mucosa by means of immunofluorescence technique as follows: (a) the FOXP3 protein (green), (b) CD3 (red), (c) nuclear stain with 4′,6 diamidino-2-phenylindole (DAPI: blue), and (d) The overlay images. The immunoreactivity for FOXP3 was significantly detected in nucleus of some CD3 positive cells. Scale bar = 100 μm.

Figure 2

Identification of the cells expressing T-bet and GATA-3 protein in human nasal mucosa. (a) The dual staining for CD3 (red) and T-bet (green) in allergic nasal mucosa. The arrows indicate the cell expressing T-bet in nucleus of CD3 positive cells. (b) The dual staining for CD3 (red) and GATA-3 (green) in allergic nasal mucosa. The arrows indicate the cell expressing GATA-3 in nucleus of CD3 positive cells. Scale bar = 100 μm.

Figure 3

The number of FOXP3+CD3+ cells in allergic and nonallergic nasal mucosa by means of double immunofluorescence staining for FOXP3 protein and CD3. There are no significantly differences between the groups in terms of the number of FOXP3+CD3+ cells.

Figure 4

The percentage of T-bet-, GATA3-, or FOXP3-positive T lymphocytes in allergic (n = 14) and nonallergic (n = 12) nasal mucosa. Sections were stained by double immunohistochemistry (IHC) using anti-CD3 antibody and antibody against each transcription factor protein. The percentage of double-positive cells to IHC-positive cells was calculated. Data are the mean ± SD.

Figure 5

The correlation between the percentage of FOXP3-positive T lymphocytes and Th1/Th2 ratio in allergic nasal mucosa (n = 14). Linear regression analysis was used to determine the relationship between the percentage of FOXP3-positive cell in CD3-positive cells and the ratio of GATA-3+CD3+ cells/T-bet+ CD3 cells in allergic nasal mucosa.