Allergy Asthma Immunol Res.  2017 Mar;9(2):142-151. 10.4168/aair.2017.9.2.142.

Vγ1+γδT Cells Are Correlated With Increasing Expression of Eosinophil Cationic Protein and Metalloproteinase-7 in Chronic Rhinosinusitis With Nasal Polyps Inducing the Formation of Edema

• Yang LY
• Li X
• Li WT
• Huang JC
• Wang ZY
• Huang ZZ
• Chang LH
• Zhang GH

• KMID: 2413388
• DOI: http://doi.org/10.4168/aair.2017.9.2.142

Abstract

PURPOSE
We have found that expression of γδT cells is increased in pathological mucosa of chronic rhinosinusitis with nasal polyps (CRSwNP) compared with normal nasal mucosa. This increase is correlated with the infiltration of eosinophils in CRSwNP. Here, we investigated the expression of γδT cells, inflammation and tissue remodeling factors as well as their probable relationships in different types of chronic rhinosinusitis (CRS) in China.
METHODS
A total of 76 surgical tissue samples that included 43 CRSwNP samples (15 eosinophilic and 28 non-eosinophilic), 17 CRS samples without nasal polyps (CRSsNP), and 16 controls were obtained. Quantitative reverse transcription-polymerase chain reaction (RT-PCR) was used to measure the mRNA expression levels of Vγ1⁺γδT cells, Vγ4⁺γδT cells, eosinophil cationic protein (ECP), interleukin (IL)-8, transforming growth factor (TGF)-β2, metalloproteinase (MMP)-7, tissue inhibitor of metalloproteinase (TIMP)-4 and hypoxia-inducible factor (HIF)-1α. Enzyme linked immunosorbent assay (ELISA) was used to measure the protein level of ECP and MMP-7 in CRSwNP. The eosinophils were counted and the level of edema was analyzed with HE staining.
RESULTS
The mRNA expression levels of the Vγ1 subset, ECP and MMP-7 were significantly increased in CRSwNP with histological characteristics of eosinophilic infiltration and edema. The expression of the Vγ1 gene in CRSwNP correlated positively with the expression of both ECP and MMP-7. No significant decreases in the mRNA expression levels of TGF-β2, TIMP-4 or HIF-1α were observed in the CRSwNP samples. The expression levels of Vγ1 gene, ECP and MMP-7 were significantly increased in eosinophilic CRSwNP compared to non-eosinophilic CRSwNP.
CONCLUSIONS
Our results suggest the associations between Vγ1⁺γδT cells, ECP and MMP-7 in CRSwNP, indicating that Vγ1⁺γδT cells can induce the eosinophilic inflammation, which has a further effect on the formation of edema.

Keyword

Chronic rhinosinusitis; nasal polyps; γδT cells; inflammation; tissue remodeling

MeSH Terms

China
Edema*
Enzyme-Linked Immunosorbent Assay
Eosinophil Cationic Protein*
Eosinophils*
Inflammation
Interleukins
Mucous Membrane
Nasal Mucosa
Nasal Polyps*
RNA, Messenger
Transforming Growth Factors
Eosinophil Cationic Protein
Interleukins
RNA, Messenger
Transforming Growth Factors

Figure

• 

  Fig. 1 The relative mRNA expression levels for Vγ1+ κ cells, Vγ4+ κ cells, ECP, IL-8, TGF-β2, MMP-7, TIMP-4, and HIF-1α.

• 

  Fig. 2 A large amount of eosinophil infiltration in the subepithelial tissue of a CRSwNP sample compared to CRSsNP and control (HE-stained section). (A) CRSwNP: epithelial cell damage and basement membrane thickness (200× magnification). (B) CRSwNP: eosinophils scattered in the subepithelial tissue with pseudocyst formation and edema, a lack of collagen and the disappearance of blood vessels and glands (400× magnification). (C) CRSsNP: lymphocytes and neutrophils scattered with increased collagen and fibrosis (400× magnification). (D) Control: normal nasal mucosa (400× magnification of a control sample).

• 

  Fig. 3 Semiquantitative edema scores and eosinophil infiltrations in the different CRS types and controls. (A) Edema: the semiquantitative score of the severity of edema. (B) Eosinophil: the percentage of eosinophils relative to the total inflammatory cells.

• 

  Fig. 4 Hellquist's classification of CRS. HE-stained tissue sections at 200× magnification. (A) Edematous type: subepithelial edema, a lack of collagen, and the disappearance of glands and blood vessels with inflammatory cell infiltration. (B) Fibrotic type: thickened basement membrane, subepithelial collagen deposition, and inflammatory cell infiltration. (C) Glandular hyperplastic type: increased amounts of glands and blood vessels.

• 

  Fig. 5 Correlations between κ cells, inflammation and remodeling. (A) The correlation coefficient for the relationship of Eos and ECP: rs=0.386, 0.011≤p<0.05. (B) The correlation coefficient for the association between MMP-7 and ECP: rs=0. 418, 0.030≤p<0.05. (C) The correlation coefficient for the association between ECP and Vγ1+ κ cells: rs=0.368, 0.019≤p<0.05. (D) The correlation coefficient for the association between MMP-7 and Vγ1+ κ cells: rs=0.353, 0.032≤ p<0.05. The data (x) regarding the relative mRNA expression levels of ECP, MMP-7 and Vγ1+ κ cells were transformed with same equation {x′=log10 (105x+1)}.

• 

  Fig. 6 The relative mRNA expression levels for (A) Vγ1+ κ cells, (B) the protein level of ECP, and (C) MMP-7 in Eos CRSwNP compared to non-Eos CRSwNP.

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