Immunopathologic Characteristics Of Different Types Of CRS And The Involved Mechanisms

Objective:CRSwNP and CRSsNP is reported to be different in inflammatory patterns of the sinonasal mucosa in white patients, the former displays Th2 skewed eosinophilic inflammation while the latter presents Th1 dominant inflammation. Studies in nonwhite populations may further be helpful to understand the pathogenic mechanisms of CRS. The present study, firstly, is to investigate the immunopathologic profiles of CRSwNP and CRSsNP in adult Chinese.Methods:Histologic characteristics of surgical samples were analyzed through HE stain in 50 controls,94 CRSsNP patients, and 151 CRSwNP patients. CRSsNP and CRSwNP were divided into eosinophilic CRSsNP and CRSwNP, non-eosinophilic CRSsNP and CRSwNP according to the percentage of eosinophils out of total inflammatory cells. Tissue samples from 17 controls,36 CRSsNP patients, and 45 CRSwNP patients were stained for CD3, CD4, CD8, CD20, CD68, myeloperoxidase (MPO), and dendritic cell lysosomeassociated membrane protein (DC-LAMP). Expression profiles of transcription factors of T-cell subsets in relation to cytokines were examined by means of quantitative RT-PCR.Results:Over half of CRSwNP patients presented noneosinophilic inflammation. CRSwNP had a higher number of eosinophils, plasma cells, and CD3+, CD8+, CD20+, and CD68+ cells and a lower MPO expression rate than CRSsNP. Expression levels of transcription factors and cytokines of Th1/Th2/Th17 were increased whereas the mRNA expression rate of FOXP3 and TGF-β1 was decreased in both CRSsNP and CRSwNP compared with controls. Comparing CRSsNP and CRSwNP, CRSsNP had higher levels of IFN-γexpression, whereas only eosinophilic CRSwNP demonstrated an enhanced expression of GATA-3 and IL-5. Compared with noneosinophilic CRSwNP, an exaggerated Th2/Th17 reaction was found in eosinophilic CRSwNP.Conclusion:Both Chinese CRSsNP and CRSwNP patients demonstrate impaired regulatory T cell function and enhanced Th1/Th2/Th17 responses. CRSsNP is confirmed to be a predominant Th1 milieu, whereas Th2 skewed inflammation with predominant Th17 reactions can be demonstrated only in eosinophilic CRSwNP, but not in noneosinophilic CRSwNP. Objective:We found that Th2-skewed eosinophilic inflammation and enhanced Th17 reaction existed only in Eos-CRSwNP but not non-Eos-CRSwNP, however, compared with Eos-CRSwNP, non-Eos CRSwNP had higher frequency of neutrophil infiltration, which suggests different pathogenesis between Eos-CRSwNP and non-Eos-CRSwNP and that distinct T helpers reaction in them may contribute to it. So the purposes of the present study were to 1. investigate the activation of eosinophils and neutrophils in Eos-CRSwNP and non-Eos-CRSwNP,2. measure Thl/Th2/Thl7 cytokines in protein level and their corresponding chemotactic chemokines and receptors of the chemotactic chemokines,3. compare the clinical manifestation between Eos-CRSwNP and non-Eos-CRSwNP,4. explore what results in the different immunopathologic feasures between Eos-CRSwNP and non-Eos-CRSwNP.Methods:First we divided CRSwNP into Eos-CRSwNP and non-Eos-CRSwNP according to the standard set up in our previous paper, and compared their clinical menifestation. Immunohistochemistry was used to detect active eosinophils (MBP+, major basic protein,) and neutrophils (MPO+, myeloperoxidase). Real time PCR or Bio-plex protein detecting technique was used to measure eosinophil chemotactic factors, total eotaxin,RANTES and GM-CSF, and neutrophil chemotactic factors, IL-8 and GRO-a in the homogenate of control,Eos-CRSwNP and non-Eos-CRSwNP tissue. Th1/Th2/Th17 cytokines IFN-gamma,IL-4,IL-5,IL-13 and IL-17 protein level were tested by bio-plex technique, Th1,Th2 chemotactic factors IP-10,TARC and MDC were detected by ELISA, the receptors of Thl/Th2/Th17 chemotactic factors CXCL3,CCR4,CCR6 and Th1 attracting factors MIG,I-TAC were detected by Real time PCR. Peripheral blood and tissue homogenate total IgE,specific IgE to air-borne allergen and tissue homogenate specific IgE to Staphylococcus enterotoxin were detected by Immunocap.Results:Of 39 patients with CRSwNP,16 subjects (41%) displayed eosinophilic inflammation, while 23 subjects (59%) showed non-eosinophilic inflammation, no clinical manifestation difference was found between them. Compared with control and non-Eos-CRSwNP, active eosinophils (MBP+) were increased in Eos-CRSwNP and correlated with eosinophilic inflammation. Eos-CRSwNP had higher eotaxin expression but not RANTES and GM-CSF compared with control and non-Eos-CRSwNP. Though non-Eos-CRSwNP had higher frequency of neutrophil infiltration, but some Eos-CRSwNP also presented prominent neutrophilic inflammation. IL-8 expression was enhanced both in Eos-CRSwNP and non-Eos-CRSwNP when compared with control, whereas the expression of another neutrophil attracting factor GRO-awas higher in non-Eos-CRSwNP than control and Eos-CRSwNP. Local total IgE, CCR6 (receptor expressed by Th17), Th2 cytokines IL-5,IL-13,Th2 attracting factor TARC and CCR4 (the receptor expressed by Th2 cells) were increased in Eos-CRSwNP compared with control and non-Eos-CRSwNP. Compared with control, the expression of Thl cytokines IFN-γ, CXCR3 (the receptor of Th1 attracting factors) were enhanced in Eos-CRSwNP but not non-Eos-CRSwNP. Specific IgE to Staphylococcus enterotoxin A and B was only found in 30% Eos-CRSwNP while in none of non-Eos-CRSwNP and control. Local total IgE was correlated positively with local eosinophilic inflammation and Th2 cytokine IL-13 expression.Conclusion:Different immunopathologic characteristics between Eos-CRSwNP and non-Eos-CRSwNP was further confirmed:Th2-skewed and enhanced Th17 immunity reaction only exist in Eos-CRSwNP but not non-Eos-CRSwNP, non-Eos-CRSwNP has higher frequency of neutrophil infiltration and neutrophil activation. Local IgE may contribute to the formation of different CRSwNP phenotype and staphylococcus superantigen may only play a role in partial Eos-CRSwNP. Th2 immunity cells and eosinophilils in Eos-CRSwNP may be recruited by TARC and eotaxin respectively, while GRO-a may play an important role in neutrophil trafficing and activation in non-Eos-CRSwNP. Objective:To observe the pathologic characteristics, and investigate mast cell and its activation in Chronic rhinosinusitis (CRSsNP) and their relations with eosinophilic inflammation. Methods:HE stain was used to observe tissue features and count total inflammatory cells, mononuclear cells, plasma cells and eosinophilils in lamina propria of CRSsNP and inferior turbinate. Toluidine blue stain and immunohistochemical stain for tryptase were used to detect mast cell and its activation respectively in CRSsNP and control, and their corelations with tissue eosinophilia were analysed. Results:CRSsNP has increased total inflammatory cells, mononuclear cells and plasma cells but comparable eosinophilils and lamina propria glands compared with inferior turbinate. Mast cells corelated with activated mast cells, but there was no difference between CRSsNP and control for both of them and there were no corelation between mast cell and its activation with tissue eosinophilia. Conclusion:CRSsNP has more serious inflammation but no more mast cell and its activation and eosinophil compared with inferior turbinate, and there were no corelations between mast cell and its activation with eosinophil count which suggests that mast cell and eosinophic inflammation mediated by it may not play an important role in the pathogenesis of CRSsNP.