Allergy Asthma Immunol Res.  2012 Jan;4(1):24-30. 10.4168/aair.2012.4.1.24.

Decreased Expression of FOXP3 in Nasal Polyposis

• Roongrotwattanasiri K
• Pawankar R
• Kimura S
• Mori S
• Nonaka M
• Yagi T

• KMID: 2130375
• DOI: http://doi.org/10.4168/aair.2012.4.1.24

Abstract

PURPOSE
The pathogenesis of nasal polyposis (NP) is unclear. Eosinophils and mast cells are considered to play important roles in this process. In addition, the levels of Th2-type cells are increased, irrespective of the atopic status of the patient with NP. In this context, we and others have shown high levels of thymus and activation-related chemokine/CCL17, macrophage-derived chemokine, eotaxin, and RANTES in patients with NP. Forkhead box P3 (FOXP3) plays a key role in CD4+CD25+ regulatory T-cell function and represents a specific marker for regulatory T cells (Tregs). Decreased expression of FOXP3 has been reported in allergic diseases. The present study was designed to evaluate the presence and potential roles of Tregs, defined by the expression of FOXP3 protein, in NP.
METHODS
Using immunohistochemistry, we estimated the numbers of FOXP3+ cells in the epithelium and lamina propria of the NPs of 17 patients with chronic rhinosinusitis with NP and the nasal mucosa of 15 patients with allergic rhinitis (AR). The number of FOXP3+ cells in NPs was compared with that in the nasal mucosa of patients with AR, and the numbers of FOXP3+ cells in atopic and non-atopic NP were also compared.
RESULTS
The number of FOXP3+ cells in the lamina propria of patients with NP was significantly lower than that in the nasal mucosa of the AR patients (2.79 vs. 5.99, P=0.008). There was no statistically significant difference noted for the numbers of FOXP3+ cells between the epithelium of the NP and the nasal mucosa (3.60 vs. 2.39, P=0.180). Furthermore, the numbers of CD4+FOXP3+ cells were lower in NPs than in the allergic nasal mucosa. There was no difference in the number of FOXP3+ cells between the atopic and non-atopic NP patients.
CONCLUSIONS
Fewer Tregs (i.e., decreased FOXP3 expression) are found in NPs than in the nasal mucosa of AR patients. As the severity of eosinophilic, Th2-type inflammation and the levels of inflammatory mediators are much higher in NPs than in the nasal mucosa of AR patients, an inverse co-relationship may exist between these parameters and the number of Tregs. The deficiency of Tregs in NP may account for the more pronounced Th2-type inflammation seen in these patients.

Keyword

Nasal polyposis; FOXP3; regulatory T cells; Th2; inflammation; IgE

MeSH Terms

Chemokine CCL22
Chemokine CCL5
Eosinophils
Epithelium
Humans
Immunoglobulin E
Immunohistochemistry
Inflammation
Mast Cells
Mucous Membrane
Nasal Mucosa
Rhinitis
Rhinitis, Allergic, Perennial
T-Lymphocytes
T-Lymphocytes, Regulatory
Thymus Gland
Chemokine CCL22
Chemokine CCL5
Immunoglobulin E

Figure

• 

  Fig. 1 Immunoreactivity and localization of forkhead box P3 (FOXP3) protein in nasal polyps. The immunoreactivity for FOXP3 was analyzed by immunohistochemistry using the peroxidase-based avidin-biotin complex method, as described in the text. FOXP3 immunoreactivity is shown in (A), with FOXP3 expression being detected mainly in the immune cells of the lamina propria. (B) The negative control shows no immunoreactivity for FOXP3. Magnification ×400 HPF.

• 

  Fig. 2 Immunoreactivity and localization of forkhead box P3 (FOXP3) in the nasal mucosa of patients with allergic rhinitis. Immunoreactivity for FOXP3 was analyzed by immunohistochemistry using the peroxidase-based avidin-biotin complex method, as described in the text. FOXP3 immunoreactivity is shown in (A), with many FOXP3-expressing cells being evident in the lamina propria. (B) The negative control shows no immunoreactivity for FOXP3. Magnification ×400 HPF.

• 

  Fig. 3 Immunoreactivity and localization of forkhead box P3 (FOXP3)+CD4+ cells. Double-staining using labeled streptavidin biotin (LSAB) detection with 3,3'-diaminobenzidine (DAB-brown) for FOXP3 expression and LSAB alkaline phosphatase detection with Fast Red substrate (red) for CD4+ cells was performed as described in the text (A). Symbols used: ☆, FOXP3+/CD4+ cell; △, FOXP3-/CD4+ cell. (B) Double-staining with anti-FOXP3 mAb and IgG1 (negative control for CD4). The arrow indicates FOXP3+ cells. Immunoreactivity is evident for only FOXP3+ cells. There is no immunoreactivity for IgG1. (C) The negative control for both FOXP3 and CD4 shows no immunoreactivity for FOXP3 or CD4. Magnification ×400 HPF

• 

  Fig. 4 (A) The numbers of forkhead box P3 (FOXP3)-positive cells in the epithelia and lamina propria of nasal polyps. Immunoreactivity for FOXP3 was analyzed by immunohistochemistry using the peroxidase-based avidin-biotin complex (ABC) method, as described in the text. Positively stained cells were counted as described in the text. Magnification ×400 HPF. There is no statistically significant difference between the numbers of FOXP3+ cells in the epithelia and lamina propria. (B) The numbers of FOXP3+ cells in the epithelia and lamina propria of the nasal mucosa of patients with allergic rhinitis. Immunoreactivity for FOXP3 was analyzed by immunohistochemistry using the peroxidase-based ABC method, as described in the text. Positively stained cells were counted as described in the text. Magnification ×400 HPF. The number of FOXP3+ cells is significantly higher in the lamina propria than in the epithelium. *P<0.01.

• 

  Fig. 5 (A) Comparison of the numbers of forkhead box P3 (FOXP3)-positive cells in the lamina propria of nasal polyps and allergic nasal mucosa. Immunoreactivity for FOXP3 was analyzed by immunohistochemistry using the peroxidase-based avidin-biotin complex (ABC) method, as described in the text. Positively stained cells were counted as described in the text. Magnification ×400 HPF. The number of FOXP3+ cells is significantly lower in the lamina propria of the nasal polyps than in those of the allergic nasal mucosa. *P<0.01. (B) Comparison of the numbers of FOXP3+ cells in the epithelia of nasal polyps and allergic nasal mucosa. Immunoreactivity for FOXP3 was analyzed by immunohistochemistry using the peroxidase-based ABC method, as described in the text. Positively stained cells were counted as described in the text. Magnification ×400 HPF. Although the number of FOXP3+ cells is higher in the epithelia of the allergic nasal mucosa than in those of the nasal polyps, the difference is not statistically significant (NS).

• 

  Fig. 6 Comparison of the numbers of forkhead box P3 (FOXP3)-positive cells in the epithelia of nasal polyps and allergic nasal mucosa. Double-staining for FOXP3 and CD4 was performed as described in the text. The number of FOXP3+CD4+ cells in the nasal mucosa of allergic rhinitis patients is significantly higher than that in the nasal polyps. *P<0.01.

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