The Journal of Allergy and Clinical Immunology
Volume 125, Issue 2 , Pages 507-508, February 2010

Reply

  • Ping-Ping Cao, MD

      Affiliations

    • Department of Otolaryngology-Head and Neck Surgery, Tongji Hospital, Tongji Medical College, Huazhong University of Science and Technology, Wuhan, PR China
    • ,
  • Hua-Bin Li, MD, PhD

      Affiliations

    • Otolaryngology Hospital, the First Affiliated Hospital of Sun Yat-sen University, Guangzhou, PR China
    • ,
  • De-Yun Wang, MD, PhD

      Affiliations

    • Department of Otolaryngology, Yong Loo Lin School of Medicine, National University of Singapore
    • ,
  • Martin Desrosiers, MD

      Affiliations

    • Department of Otolaryngology-Head and Neck Surgery, Universite de Montreal, Quebec, Canada
    • Department of Otolaryngology-Head and Neck Surgery, McGill University, Montreal, Quebec, Canada
    • ,
  • Zheng Liu, MD, PhD

      Affiliations

    • Department of Otolaryngology-Head and Neck Surgery, Tongji Hospital, Tongji Medical College, Huazhong University of Science and Technology, Wuhan, PR China

published online 11 January 2010.

Article Outline

 

To the Editor:

We appreciate the critical comments from Zhang et al1 on our study.2 The major criticism questions the sensitivity and accuracy of the measurements of TGF-β1 and forkhead box protein 3 (Foxp3) mRNA expression in our study,2 because these results are discordant with previous reports by their group.1, 3, 4 In response to this question, we have retested TGF-β1 and Foxp3 mRNA expression, using both the primers constructed for our previous study2 and new ones constructed from the sequences reported by their group3, 4 in a new set of samples obtained from 10 patients with septal deviation (controls) and 14 patients with chronic rhinosinusitis without nasal polyps (CRSsNP). With regard to TGF-β1, with our primers, we could again only identify positive expression in a limited number of samples, for an expression frequency of 50% in CRSsNP and 70% in controls, which is comparable to our previous results.2 Using the primers constructed according to the description by their group,3, 4 we were able to detect the expression of TGF-β1 in all samples, but the expression intensity was relatively weak. Such discrepancy might be a result of the different amplification efficiencies between different primers.5 Furthermore, when using the primers of their group, TGF-β1 mRNA expression levels were almost 2-fold lower in CRSsNP compared with controls, which again confirms our previous findings,2 but not those reported by Zhang et al.1

As for Foxp3, whether using our primers or the primers of their group, we again did not detect Foxp3 mRNA expression in all of the samples, and the same expression frequencies (CRSsNP vs controls: 50% vs 70%) were found, which is also consistent with our previous report.2

The discrepancies noted between our results and those of Zhang et al1, 3 are not unique. Contradictory results of TGF-β1 and Foxp3 expression levels in both healthy and inflamed nasal tissues have been reported in the literature.6

Our classification of chronic rhinosinusitis with nasal polyps (CRSwNP) into eosinophilic and noneosinophilic subgroups on the basis of eosinophil count in the polyp tissue,2 a commonly used histologic assessment, was questioned by Zhang et al1 because they assumed that eosinophils identified in CRSwNP samples may not be activated.3 However, we believe that measuring eosinophilic cationic protein levels in tissue homogenates as described in the study by Zhang et al3 may be not a reliable way to confirm the activation state of eosinophils.7 First, eosinophilic cationic protein is found not only in eosinophils but also in neutrophils.8 Second, such measurement is unable to differentiate between secreted (an indicator of cell activation) and intracellular eosinophilic cationic protein. In additional support of our classification method, using our classification criteria, 2 distinct types of CRSwNP with different immunopathologic phenotypes were revealed.2

With respect to criticisms regarding statistical analysis, although 5 groups were included in Fig 1 in our article,2 the comparisons were made only among controls, CRSsNP, and CRSwNP, and among controls, CRSsNP, and noneosinophilic and eosinophilic CRSwNP.

We agree with the comment that our mRNA data need to be confirmed at the protein level. Nevertheless, the weak expression intensity of these markers may represent a significant obstacle to reliable quantitative analysis. To attain a definitive conclusion about TGF-β1 and Foxp3 in chronic rhinosinusitis, we believe not only that more sensitive protein detection techniques should be used, but also that more proteins need to be assessed in what is obviously a highly complex network.

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References 

  1. Zhang N, Li S, Lin P, Li X, van Bruaene N, Zhang J, et al. Remodeling and inflammation in Chinese versus white patients with chronic rhinosinusitis. J Allergy Clin Immunol. 2010;125:507
  2. Cao PP, Li HB, Wang BF, Wang SB, You XJ, Cui YH, et al. Distinct immunopathologic characteristics of various types of chronic rhinosinusitis in adult Chinese. J Allergy Clin Immunol. 2009;124:478–484
  3. Zhang N, Van Zele T, Perez-Novo C, Van Bruaene N, Holtappels G, DeRuyck N, et al. Different types of T-effector cells orchestrate mucosal inflammation in chronic sinus disease. J Allergy Clin Immunol. 2008;122:961–968
  4. Van Bruaene N, Perez-Novo C, Basinski T, Van Zele T, Holtappels G, Schmidt-Weber C, et al. T cell regulation in chronic paranasal sinus disease. J Allergy Clin Immunol. 2008;121:1435–1441
  5. Liu W, Saint DA. A new quantitative method of real time reverse transcription polymerase chain reaction assay based on stimulation of polymerase chain reaction kinetics. Anal Biochem. 2002;302:52–59
  6. Peters AT, Kato A, Carr T, Suh L, Norton J, Grammer LC, et al. Analysis of the Th17 pathway in chronic rhinosinusitis. J Allergy Clin Immunol. 2009;123 (Suppl 2):145
  7. Khan DA, Cody DT, George TJ, Gleich GJ, Leiferman KM. Allergic fungal sinusitis: an immuohistologic analysis. J Allergy Clin Immunol. 2000;106:1096–1101
  8. Sur S, Glitz DG, Kita H, Kujawa SM, Peterson EA, Weiler DA, et al. Localization of eosinophil-derived neurotoxin and eosinophil cationic protein in neutrophilic leukocytes. J Leukoc Biol. 1998;63:715–722

 Disclosure of potential conflict of interest: The authors have declared that they have no conflict of interest.

PII: S0091-6749(09)01543-7

doi:10.1016/j.jaci.2009.10.016

Refers to article:

  • Remodeling and inflammation in Chinese versus white patients with chronic rhinosinusitis , 11 January 2010

    Nan Zhang, Shixi Liu, Ping Lin, Xiaoyuan Li, Nicholas van Bruaene, Jie Zhang, Thibaut van Zele, Claus Bachert
    The Journal of Allergy and Clinical Immunology February 2010 (Vol. 125, Issue 2, Page 507)

The Journal of Allergy and Clinical Immunology
Volume 125, Issue 2 , Pages 507-508, February 2010

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