Turbinate-homing IgA-secreting cells originate in the nasal lymphoid tissues.

Nasal vaccination elicits a humoral immune response that provides protection from airborne pathogens1, yet the origins and specific immune niches of antigen-specific IgA-secreting cells in the upper airways are unclear2. Here we define nasal glandular acinar structures and the turbinates as immunological niches that recruit IgA-secreting plasma cells from the nasal-associated lymphoid tissues (NALTs)3. Using intact organ imaging, we demonstrate that nasal vaccination induces B cell expansion in the subepithelial dome of the NALT, followed by invasion into commensal-bacteria-driven chronic germinal centres in a T cell-dependent manner. Initiation of the germinal centre response in the NALT requires pre-expansion of antigen-specific T cells, which interact with cognate B cells in interfollicular regions. NALT ablation and blockade of PSGL-1, which mediates interactions with endothelial cell selectins, demonstrated that NALT-derived IgA-expressing B cells home to the turbinate region through the circulation, where they are positioned primarily around glandular acinar structures. CCL28 expression was increased in the turbinates in response to vaccination and promoted homing of IgA+ B cells to this site. Thus, in response to nasal vaccination, the glandular acini and turbinates provide immunological niches that host NALT-derived IgA-secreting cells. These cellular events could be manipulated in vaccine design or in the treatment of upper airway allergic responses.

Antiviral activity of bovine type III interferon against bovine viral diarrhea virus is greatly reduced in bovine turbinate cells due to limited expression of IFN lambda receptor 1 (IL-28Rα).

Introduction: The antiviral activity of recombinant bovine interferon lambda 3 (bovIFN-λ3) against bovine viral diarrhea virus (BVDV) has been demonstrated in vitro in Madin-Darby bovine kidney cells (MDBK) and in vivo in cattle. However, anti-BVDV activity of bovIFN-λ3 has not been studied in bovine respiratory tract epithelial cells, supposedly a primary target of BVDV infection when entering the host by the oronasal route. Methods: Here we investigated the anti-BVDV activity of bovIFN-λ3 in bovine turbinate-derived primary epithelial cells (BTu) using BVDV infection and immunoperoxidase staining, TCID50, RT-qPCR, DNA and transcriptome sequencing, and transfection with plasmids containing the two subunits, IL-28Rα and IL-10Rß that constitute the bovIFN-λ3 receptor. Results: Our immunoperoxidase staining, RT-qPCR, and TCID50 results show that while BVDV was successfully cleared in MDBK cells treated with bovIFN-λ3 and bovIFN-α, only the latter, bovIFN-α, cleared BVDV in BTu cells. Preincubation of MDBK cells with bovIFN-λ3 before BVDV infection was needed to induce optimal antiviral state. Both cell types displayed intact type I and III IFN signaling pathways and expressed similar levels of IL-10Rß subunit of the type III IFN receptor. Sequencing of PCR amplicon of the IL-28Rα subunit revealed intact transmembrane domain and lack of single nucleotide polymorphisms (SNPs) in BTu cells. However, RT-qPCR and transcriptomic analyses showed a lower expression of IL-28Rα transcripts in BTu cells as compared to MDBK cells. Interestingly, transfection of BTu cells with a plasmid encoding IL-28Rα subunit, but not IL-10Rß subunit, established the bovIFN-λ3 sensitivity showing similar anti-BVDV activity to the response in MDBK cells. Conclusion: Our results demonstrate that the sensitivity of cells to bovIFN-λ3 depends not only on the quality but also of the quantity of the IL-28Rα subunit of the heterodimeric receptor. A reduction in IL-28Rα transcript expression was detected in BTu as compared to MDBK cells, despite the absence of spliced variants or SNPs. The establishment of bovIFN-λ3 induced anti-BVDV activity in BTu cells transfected with an IL-28Rα plasmid suggests that the level of expression of this receptor subunit is crucial for the specific antiviral activity of type III IFN in these cells.

The role of nasal cytology in the management of inferior turbinate hypertrophy.

Inferior turbinate hypertrophy (ITH) is the main cause of nasal obstruction symptom. This study aimed at investigating whether a particular cellular pattern could be a predictive factor for failure of medical treatment for ITH in patients with rhinitis. Globally, 258 patients with chronic nasal obstruction due to ITH were evaluated by: visual analogue scale assessment of symptoms, skin prick tests, fiber-endoscopy, active anterior rhinomanometry, and nasal cytology. All patients were treated with drugs for 3 months and then re-evaluated. The symptom improvement depended on the different cellular pattern. There was improvement in: 54 (51.4 percent) patients with allergic rhinitis, 72 (69.2 percent) with non-allergic rhinitis with neutrophils (NARNE), 15 (42.8 percent) with non-allergic rhinitis with eosinophils (NARES), and 9 (64.3 percent) with non-allergic rhinitis with mast cells/non-allergic rhinitis with eosinophils and mast cells (NARMA/NARESMA). The non-responders (108; 41.9 percent) were therefore directed towards surgical treatment. Both patients with allergic rhinitis and patients affected by NARES had a higher failure rate to medical treatment compared with NARMA and NARESMA groups (pless than0.01). In conclusion, elevated number of eosinophils, in the nasal secretion of both allergic (allergic rhinitis) and non-allergic (NARES) patients with ITH, can be associated to a higher medical treatment failure rate.

The airway antigen sampling system: respiratory M cells as an alternative gateway for inhaled antigens.

In this study, we demonstrated a new airway Ag sampling site by analyzing tissue sections of the murine nasal passages. We revealed the presence of respiratory M cells, which had the ability to take up OVA and recombinant Salmonella typhimurium expressing GFP, in the turbinates covered with single-layer epithelium. These M cells were also capable of taking up respiratory pathogen group A Streptococcus after nasal challenge. Inhibitor of DNA binding/differentiation 2 (Id2)-deficient mice, which are deficient in lymphoid tissues, including nasopharynx-associated lymphoid tissue, had a similar frequency of M cell clusters in their nasal epithelia to that of their littermates, Id2(+/-) mice. The titers of Ag-specific Abs were as high in Id2(-/-) mice as in Id2(+/-) mice after nasal immunization with recombinant Salmonella-ToxC or group A Streptococcus, indicating that respiratory M cells were capable of sampling inhaled bacterial Ag to initiate an Ag-specific immune response. Taken together, these findings suggest that respiratory M cells act as a nasopharynx-associated lymphoid tissue-independent alternative gateway for Ag sampling and subsequent induction of Ag-specific immune responses in the upper respiratory tract.

Comparison of the histologic changes in conchae induced by radiofrequency thermal ablation and submucosal diathermy.

Objective of study was to determine the histological change induced in the conchae by submucosal diathermy and radiofrequency thermal ablation, two techniques used in the treatment of lower conchal hypertrophy, and to compare the two methods to each other. The study was performed on 15 rabbits. Radiofrequency was applied to the study animals in Group I (n = 5) and submucosal diathermy to Group II (n = 5), while Group III (n = 5) was the untreated control. The animals were decapitated 21 days after treatment and their conchae nasales ventrales excised on both sides. Histology slides were prepared and evaluated by light microscopy for ciliary loss, increase in submucosal vascularity, loss of goblet cells, inflammatory cellular infiltration, fibrosis and epithelial damage. The differences between Groups I and III were not significant regarding ciliary loss, increase in submucosal vascularity, loss of goblet cells and epithelial damage (p > 0.05), while the inflammatory cellular infiltration and fibrosis were significantly different between these groups (p < 0.05). As for the differences between Groups II and III, they were significant for each of the compared parameters (p < 0.05), while among Groups I and II they were significant for ciliary loss (p < 0.05), increase in submucosal vascularity, loss of goblet cells, inflammatory cellular infiltration and epithelial damage but not fibrosis (p > 0.05). Based on these findings, we can state that the use of radiofrequency thermal ablation causes less change in the normal conchal histology than submucosal diathermy application.

Antigen-specific activities of CD8+ T cells in the nasal mucosa of patients with nasal allergy.

BACKGROUND: The prevalence of chronic rhinitis is increasing rapidly. Its pathogenesis is not fully understood but immune inflammation is one plausible causative factor. Antigen specific CD8+ T cells play a critical role in the induction of chronic inflammation. This study aims to investigate the role of antigen specific CD8+ T cells in the pathogenesis of chronic AR. METHODS: Nasal mucosal epithelial samples obtained by the surface of the nasal mucosaof patients with AR complicated with inferior turbinate hypertrophy. Exosomes were purified from the scratching samples and examined by immune gold electron microscopy. Cell culture models were employed to evaluate the effect of exosomes on modulating CD8+ T cell activity. RESULTS: Exosomes purified from patients with chronic AR carried microbial products, Staphylococcal enterotoxin B (SEB), and airborne antigen, Derp1. Dendritic cells pulsed by SEB/Derp1-carrying exosomes showed high levels of CD80, CD86 and the major histocompatibility class I (MHCI). Exosome-pulsed dendritic cells could induce naive CD3+ T cells to differentiate into CD8+ T cells. Upon exposure to a specific antigen, the CD8+ T cells released granzyme B and perforin and more than 30% antigen specific CD8+ T cells proliferated. CONCLUSIONS: Antigen specific CD8+ T cells play an important role in the pathogenesis of chronic AR complicated with inferior turbinate hypertrophy.

Different regulation of cigarette smoke induced inflammation in upper versus lower airways.

BACKGROUND: Cigarette smoke (CS) is known to initiate a cascade of mediator release and accumulation of immune and inflammatory cells in the lower airways. We investigated and compared the effects of CS on upper and lower airways, in a mouse model of subacute and chronic CS exposure. METHODS: C57BL/6 mice were whole-body exposed to mainstream CS or air, for 2, 4 and 24 weeks. Bronchoalveolar lavage fluid (BAL) was obtained and tissue cryosections from nasal turbinates were stained for neutrophils and T cells. Furthermore, we evaluated GCP-2, KC, MCP-1, MIP-3alpha, RORc, IL-17, FoxP3, and TGF-beta1 in nasal turbinates and lungs by RT-PCR. RESULTS: In both upper and lower airways, subacute CS-exposure induced the expression of GCP-2, MCP-1, MIP-3alpha and resulted in a neutrophilic influx. However, after chronic CS-exposure, there was a significant downregulation of inflammation in the upper airways, while on the contrary, lower airway inflammation remained present. Whereas nasal FoxP3 mRNA levels already increased after 2 weeks, lung FoxP3 mRNA increased only after 4 weeks, suggesting that mechanisms to suppress inflammation occur earlier and are more efficient in nose than in lungs. CONCLUSIONS: Altogether, these data demonstrate that CS induced inflammation may be differently regulated in the upper versus lower airways in mice. Furthermore, these data may help to identify new therapeutic targets in this disease model.

Differential expression and role of S100 proteins in chronic rhinosinusitis.

PURPOSE OF REVIEW: Immune system modulators have been under investigation to help elucidate the underlying pathophysiologies of chronic rhinosinusitis (CRS). Psoriasin (S100A7) and calgranulins (S100A8, S100A9, and S100A12) are S100 proteins that have been studied for their immune-mediating responses to pathogens within the context of CRS. This review highlights the expression patterns and proposed roles of S100 proteins in CRS with (CRSwNP) and without (CRSsNP) nasal polyps. RECENT FINDINGS: Elevated levels of S100A7 and S100A12 were measured in the sinonasal tissues of patients with CRSsNP compared with CRSwNP and controls. S100A12 expression in CRSsNP was significantly correlated to disease severity. Contrastingly, increased S100A8, S100A9, and S100A8/A9 levels were demonstrated in the nasal polyp tissues of patients with CRSwNP compared with those in inferior turbinate and uncinate tissues of patients with CRSsNP and controls. SUMMARY: The reported differential expression patterns and activities of psoriasin and calgranulins suggest that S100 proteins exert unique and concerted roles in mediating immunity in different subtypes of CRS. These studies will enable further investigations focused on understanding the immune-modulating mechanisms of S100 proteins in different inflammatory signaling pathways and disease phenotypes of CRS toward the pursuit of identifying new biomarkers and targets for improved outcomes.

Regulatory B Lymphocytes Colonize the Respiratory Tract of Neonatal Mice and Modulate Immune Responses of Alveolar Macrophages to RSV Infection in IL-10-Dependant Manner.

Respiratory syncytial virus (RSV) is the prevalent pathogen of lower respiratory tract infections in children. The presence of neonatal regulatory B lymphocytes (nBreg) has been associated with a poor control of RSV infection in human newborns and with bronchiolitis severity. So far, little is known about how nBreg may contribute to neonatal immunopathology to RSV. We tracked nBreg in neonatal BALB/c mice and we investigated their impact on lung innate immunity, especially their crosstalk with alveolar macrophages (AMs) upon RSV infection. We showed that the colonization by nBreg during the first week of life is a hallmark of neonatal lung whereas this population is almost absent in adult lung. This particular period of age when nBreg are abundant corresponds to the same period when RSV replication in lungs fails to generate a type-I interferons (IFN-I) response and is not contained. When neonatal AMs are exposed to RSV in vitro, they produce IFN-I that in turn enhances IL-10 production by nBreg. IL-10 reciprocally can decrease IFN-I secretion by AMs. Thus, our work identified nBreg as an important component of neonatal lungs and pointed out new immunoregulatory interactions with AMs in the context of RSV infection.

Cytochrome P4502A6 stability in a mini organ culture model of human nasal mucosa for genotoxicology studies as detected by flow cytometry.

Three dimensional mini organ cultures (MOCs) of human nasal turbinate epithelia have been shown to be a relevant tool in genotoxicology studies. MOCs allow repetitive or chronic exposure of cells in an organ specific mucosal architecture for an extended period of time and monitoring of possible adverse effects with, e.g., the comet assay. It is the aim to demonstrate whether the proteins of key enzymes of xenobiotic metabolism, represented by cytochrome P450 2A6 (CYP2A6), remain on a stable level for a culture period that allows repetitive or chronic exposure to xenobiotics. Culture of mini organs was performed by cutting pieces of 1 mm(3) from fresh specimens of human nasal turbinates. MOCs of five tissue donors were incubated on multi-well plates with BEBM, on days 0, 4, 7, 9, and 11 aliquots were transmitted to flow cytometric quantification of the CYP2A6 protein. The CYP2A6 protein could be demonstrated on all days of culture investigated. Interindividual differences were more pronounced on day 0 than at later stages of culture. Although there appeared to be a slight decrease over the culture period, flow cytometric analysis did not reveal a significant loss of the signals up to day 11. The present data could show a pre-requisite of metabolic competence of MOCs that is in contrast to single cell cultures. Thus, this type of organ culture provides an in vitro model suitable for the assessment of genotoxic effects of environmental pollutants mimicking the in vivo situation with target cells of carcinogens in their functional organ specific architecture.

Staphylococcus aureus enterotoxin B, protein A, and lipoteichoic acid stimulations in nasal polyps.

BACKGROUND: Increasing evidence points toward a modifying role of Staphylococcus aureus and its products in the pathogenesis of nasal polyposis. OBJECTIVE: The aim of this study was to investigate cytokine and mediator production after stimulation with S aureus-derived proteins enterotoxin B, protein A, and lipoteichoic acid in nasal polyp and control inferior turbinate tissue. METHODS: Tissue fragments were stimulated with RPMI (negative control), enterotoxin B, protein A, and lipoteichoic acid for 30 minutes and 24 hours. Supernatants were measured by multiplex for proinflammatory cytokines (IL-1beta, TNF-alpha) and T-cell and subset-related cytokines (IFN-gamma, IL-2, IL-4, IL-5, IL-8, IL-10, IL-12p70, IL-13). Histamine, TGF-beta1, cysteinyl leukotrienes, and prostaglandin D(2) were analyzed by ELISA. RESULTS: Thirty minutes of protein A stimulation resulted in a significant increase of histamine, leukotrienes, and prostaglandin D(2). Enterotoxin B stimulation over a period of 24 hours induced a significant increase of IL-1beta, TNF-alpha, IFN-gamma, IL-2, IL-4, IL-5, IL-10, and IL-13 in both groups, with this increase significantly higher in nasal polyps compared with controls. CONCLUSION: We here show that S aureus products have various effects on mucosal tissues: surface protein A induces mast cell degranulation, whereas enterotoxins induce the release of cytokines, with a T(H)2-skewed pattern in nasal polyps, supporting the stimulatory role of superantigens in the development of this inflammatory disease.

Turbinate-Specific IgE in Normal and Rhinitic Patients.

BACKGROUND: Specific immunoglobulin E (sIgE) within the nasal airway is likely to be the most ideal marker of allergic status, but little is known of the normative values in asymptomatic patients and those with rhinitis. OBJECTIVE: The aim of this study was to assess the diagnostic characteristics of inferior turbinate tissue biopsy sIgE in asymptomatic and rhinitic patients. METHODS: A diagnostic cross-sectional study was undertaken, involving patients who underwent inferior turbinate surgery with or without other surgical interventions. Inferior turbinate tissue biopsy was performed during surgery and was assessed for allergen sIgE (dust mite, grass [temperate or subtropical], and animal epithelium) using an automated immunoassay. Tissue sIgE was assessed among asymptomatic patients and those with nasal symptoms. Data were presented as median (interquartile range). A receiver operating curve was used to predict the diagnostic utility of turbinate tissue sIgE in determining allergic rhinitis. RESULTS: A total of 160 patients (41.89 ± 14.65 years, 36.9% females) were included. The median tissue sIgE concentration among the asymptomatic nonatopic group of patients was 0.09 (0.08-0.10) kUA/L and tissue sIgE > 0.10 kUA/L was determined as a positive threshold. Inferior turbinate tissue sIgE was shown to be a predictive test for allergic rhinitis (area under curve: 0.87, 95% confidence interval: 0.84-0.90) with 90% sensitivity and 89% negative predictive value. CONCLUSION: Inferior turbinate tissue biopsy sIgE is a sensitive tool to predict allergic rhinitis. The threshold value of 0.1 kUA/L corresponded well with the asymptomatic nonatopic group of patients. This method detects sIgE in the nasal mucosa and may be a useful test for allergic rhinitis in future research.

Chitosan microspheres containing Bordetella bronchiseptica antigens as novel vaccine against atrophic rhinitis in pigs.

The immune-stimulating activities of Bordetella bronchiseptica antigens containing dermonecrotoxin (BBD) loaded in chitosan microspheres (CMs) have already been reported in vitro and in vivo with a mouse alveolar macrophage cell line (RAW264.7) and mice. Therefore, this study attempted to demonstrate the successful induction of mucosal immune responses after the intranasal administration of BBD loaded in CMs (BBD-CMs) in colostrum-deprived pigs. The BBD was introduced to the CMs using an ionic gelation process involving tripolyphosphate (TPP). Colostrum-deprived pigs were then directly immunized through intranasal administration of the BBD-CMs. A challenge with a field isolate of B. bronchiseptica was performed ten days following the final immunization. The BBD-specific IgG and IgA titers, evident in the nasal wash and serum from the vaccinated pigs, increased with time (p<0.05). Following the challenge, the clinical signs of infection were about 6-fold lower in the vaccinated pigs compared with the nonvaccinated pigs. The grades for gross morphological changes in the turbinate bones from the vaccinated pigs were also significantly lower than the grades recorded for the nonvaccinated pigs (p<0.001). Therefore, the mucosal and systemic immune responses induced in the current study would seem to indicate that the intranasal administration of BBD-CMs may be an effective vaccine against atrophic rhinitis in pigs.

Submucous turbinectomy combined with posterior nasal neurectomy in the management of severe allergic rhinitis: clinical outcomes and local cytokine changes.

OBJECTIVES: Submucous resection of the inferior turbinate is one of the recommended methods to alleviate nasal symptoms in patients with severe allergic rhinitis patients in terms of postoperative results and preservation of nasal function. Posterior nasal neurectomy, recently developed by Kikawada, is a novel method to selectively cut the neural bundles out from the sphenopalatine foramen and to diminish the complaints of hypersecretion. This study was carried to examine the clinical effectiveness and changes in local cytokine levels of this combined surgical procedure. METHODS: Twenty-three patients with severe perennial allergic rhinitis underwent submucous turbinectomy combined with posterior nasal neurectomy under general anesthesia. The patients' subjective nasal symptoms were examined at each visit. The levels of interleukin-5 (IL-5), eotaxin and regulated on activation, normal T cell expressed and secreted (RANTES) in nasal lavages were measured before and 6 month after surgery. Nasal mucosa of the inferior turbinate was also obtained for histopathological examination in some cases. RESULTS: The mean symptom scores for sneeze, rhinorrhea, nasal obstruction, and total severity were all statistically decreased after surgery. Therapeutic effects continued to be apparent as long as 3 years after surgery. The mean levels of both IL-5 and eotaxin significantly decreased after surgery, but that of RANTES remained unchanged. Histopathological examination revealed that the number of inflammatory cells and nasal glands markedly reduced in lamina propria and the epithelial layer became covered with stratified columnar cells. CONCLUSION: Submucosal turbinectomy with posterior nasal neurectomy has remarkably improved subjective nasal symptoms in patients with severe allergic rhinitis on a long-term follow-up basis. The present study also demonstrates that the clinical effectiveness of the procedure is accompanied by decreases in local inflammatory cell infiltration and the related cytokine production.

Late production of CXCL8 in ruminant oro-nasal turbinate cells in response to Chlamydia abortus infection.

Chlamydia abortus is an obligate intracellular bacterium that is an important cause of ovine abortion worldwide. There are reports of abortions in cattle, but these are very rare compared to the reported incidence in sheep. The bacterium is transmitted oro-nasally and can establish a sub-clinical infection until pregnancy, when it can invade the placenta and induce an inflammatory cascade leading to placentitis and abortion. Early host-pathogen interactions could explain differential pathogenesis and subsequent disease outcome in ruminant species. In this study, we assessed the ability of sheep and cattle oro-nasal turbinate cells to sense and respond to C. abortus infection. The cells expressed toll like receptor (TLR) 2, TLR4, nucleotide oligomerization domain (NOD) 1 and NOD-like receptor pyrin domain containing 3 (NLRP3) mRNA. In response to C. abortus infection, both ovine and bovine turbinate cells produce CXCL8 mRNA and protein late in the bacterial developmental cycle, but do not produce IL-1ß or TNF-α. The UV-inactivated bacteria did not elicit a CXCL8 response, suggesting that intracellular multiplication of the bacteria is important for activating the signalling pathways. The production of innate immune cytokines from cattle and sheep turbinate cells in response to C. abortus infection was found to be largely similar.

Characteristics of Human Turbinate-Derived Mesenchymal Stem Cells Are Not Affected by Allergic Condition of Donor.

The characteristics of mesenchymal stem cells (MSCs) derived from human turbinates (hTMSCs) have not been investigated in allergic rhinitis. We evaluated the influence of allergic state of the donor on the characteristics, proliferation, and differentiation potential of hTMSCs, compared with hTMSCs derived from non-allergic patients. hTMSCs were isolated from five non-allergic and five allergic patients. The expression of toll-like receptors (TLRs) in hTMSCs was measured by FACS, and cell proliferation was measured using a cell counting kit. Cytokine secretion was analyzed using multiplex immunoassays. The osteogenic, chondrogenic, and adipogenic differentiation potentials of hTMSCs were evaluated by histology and gene expression analysis. In allergic patients, FACS analysis showed that TLR3 and TLR4 were more highly expressed on the surface of hTMSCs than TLR2 and TLR5. The proliferation of hTMSCs was not influenced by the presence of TLR priming. The expression of IL-6, IL-8, IL-12, IP-10, and RANTES was upregulated after the TLR4 priming. The differentiation potential of hTMSCs was not influenced by TLR priming. These characteristics of hTMSCs were similar to those of hTMSCs from non-allergic patients. We conclude that the allergic condition of the donor does not influence TLR expression, proliferation, or immunomodulatory potential of hTMSCs.

Nitric oxide modulation of the immune response against cholera toxin-adjuvated ovalbumin administered by the intranasal route.

Here, we studied the effect of aminoguanidine (AG) treatment, a nitric oxide synthase (NOS)-2 inhibitor, during the immune response against intranasal administration of ovalbumin (OVA) mixed with cholera toxin (CT) in BALB/c mice. NOS-2 mRNA was detected by reverse transcription-PCR (RT-PCR) in samples of lungs and turbinates early post-inoculation of the antigen. Animals intranasally treated with AG, showed an increase in the levels of seric specific IgG and IgM. A higher IgG1/IgG2a ratio against OVA was also observed in sera of same animals. Moreover, high levels of specific IgA were detected in samples of pulmonar washings obtained from treated animals. On the contrary, treated animals showed a lower DTH response while splenocytes obtained from the same animals showed a reduced proliferative capability against OVA compared to controls. Finally, RT-PCR analysis showed increased expression of TGF-beta in turbinates, lungs and cells from pulmonar washings obtained from AG treated mice. Taken together, these data suggest a role of nitric oxide (NO) in modulating the primary immune response against intranasal antigens.

Characterization of inflammatory reaction in upper airways of cystic fibrosis patients.

Inflammatory cell populations have not been yet precisely evaluated in cystic fibrosis (CF) airways. We intended to characterize morphological modifications, inflammatory cell infiltration and cell proliferation in nasal tissues obtained from 15 CF patients and from 6 non-CF patients with nasal polyposis. Morphological analysis showed an intense inflammatory infiltration in CF and non-CF tissues with only few modifications in the epithelium from CF tissues. Inflammatory cell populations characterized by specific immunolabeling were quantified, showing a predominance of macrophages and T- and B-lymphocytes and only moderate numbers of neutrophils in CF tissues; in non-CF polyps, lymphocytes and eosinophils were abundant. Proliferating cell percentages quantified after proliferating cell nuclear antigen immunolabeling were 5.3+/-4.1% (mean +/- SD) in CF polyps and 3.1+/-1.2% in non-CF polyps in epithelium but were very low in lamina propria. Intense inflammation in nasal tissues from CF patients is therefore dominated by macrophages and lymphocytes rather than by neutrophils. While morphology is preserved, proliferation is high in epithelium from CF polyps. These findings should be regarded in the future for a better understanding of inflammation in CF airway disease.

Enhancement of nasal inflammatory and epithelial responses after ozone and allergen coexposure in Brown Norway rats.

Repeated exposures to ozone cause inflammation and mucous cell metaplasia (MCM) in the nasal mucosa of laboratory animals. Similar cellular responses occur in humans during allergic rhinitis. We tested the hypothesis that exposure to ozone will enhance the inflammatory and epithelial responses associated with allergic rhinitis. Ovalbumin (OVA)-sensitized Brown Norway rats were exposed to ozone (0.5 ppm, 8 h/day) for 1 day or 3 consecutive days. Immediately after each ozone exposure, animals were challenged intranasally (IN) with either sterile saline or OVA dissolved in saline (1%, 50 microg/nasal passage). Twenty-four h after the last IN challenge rats were sacrificed; nasal tissues were removed and processed for light microscopic examination and morphometric analysis of numeric densities of inflammatory and epithelial cell populations and volume densities of intraepithelial mucosubstances. A single OVA challenge caused a significant influx of neutrophils and eosinophils into the submucosa of all nasal tissues. Ozone exposure further enhanced the appearance of eosinophils in the maxilloturbinates of OVA-challenged rats but did not increase inflammation in other nasal tissues. After 3 days of ozone/OVA coexposures, the nasal transitional epithelium lining the maxilloturbinates had increased numbers of epithelial cells as well as the appearance of mucus-containing cells in areas normally absent of these secretory cells (i.e., MCM). Multiple challenges with OVA caused increased epithelial mucosubstances in the respiratory epithelium lining the septum without increasing the number of epithelial cells. Multiple exposures to both ozone and OVA caused greater increases in intraepithelial mucosubstances in the septum than those elicited by OVA alone. These results demonstrate that exposure to ozone exacerbates epithelial and inflammatory responses associated with allergen challenge. In addition, coexposure of these agents enhanced the induced production of nasal mucosubstances caused by either agent alone.

Immunoglobulin E distribution in atopic nasal mucosa.

The distribution of B lymphocytes and immunoglobulins G, A, M, and E in nasal mucosa was studied in frozen biopsy sections of nasal turbinate from 16 allergic patients and 8 controls. The immunoperoxidase technique was used with monoclonal and polyclonal antibodies. Comparative analyses of serum immunoglobulin levels were also performed. Few B lymphocytes were observed in the nasal mucosa linings in specimens from allergic and non-allergic patients. In both groups, high positivity for IgG and IgA was observed in the nasal mucosa linings in the specimens. IgM concentration was minimal in both groups. IgE was absent in the nasal turbinate specimens of nonallergic subjects, but was present discontinuously in low concentrations in 7 of the 16 allergic patients. There was no significant difference between allergic and nonallergic patients in the tissue and serum IgG, IgA, and IgM concentrations found. IgE was detected slightly in the nasal mucosa of patients with high IgE serum concentrations (greater than 1000 IU/mL) as well as in patients with very low IgE serum concentration readings. This result raises some doubt on the hypothesis concerning the local production of IgE.