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Objectives: Investigate immune responses of adenoidal lymphocytes against outer-membrane protein P6 purified from nontypeable Haemophilus influenzae (HI). Clarify the role of adenoids in regulating the colonization of HI in the nasopharynx.Study Design: Microbiological and immunological examinations of adenoids obtained from 21 children, 15 boys and five girls, from 1 to 13 years of age (median, 5 y), suffering from adenoidal hypertrophy complicated by otitis media with effusion (OME).Methods: The incidence of HI in adenoids was compared with the number of P6-specific immunoglobulin (Ig) A-secreting cells in adenoids, determined by enzyme-linked immunoassay.Results: Quantitative culture assay showed significant correlation between the numbers of HI in adenoids and those in nasopharyngeal secretions (NS). In children aged 5 years and younger, the numbers of P6-specific IgA-secreting cells in adenoids were significantly correlated with IgA antibody titers in NS (r = 0.68, P < .05). The numbers of P6-specific IgG- and IgA-secreting cells were lower in children aged 6 years and older than in children aged 5 years and younger. Furthermore, the number of P6-specific IgA-secreting cells was significantly increased in HI-negative subjects when compared with HI-positive subjects (P < .05).Conclusions: Adenoids play an important role as an effector site of the mucosal immune system in the upper respiratory tract. IgA immune responses in adenoids are responsible for the clearance of HI from the nasopharynx. [ABSTRACT FROM AUTHOR]

Recent advances in reconstructive surgery for head and neck cancer have improved the cure rate of advanced carcinoma, and the function of the organ. However, it still remains difficult to repair the mandible and oral floor. We devised a combined flap of myocutaneous latissimus dorsi and iliac bone, and applied it to two patients with advanced carcinoma of the oral cavity that invaded to the mandible (T4N3M0). Each patient received preoperative irradiation, totalling 30 Gy and 40 Gy. Two weeks before the extensive resection, a sufficient bony mass for the presumed mandibular defect was taken from the iliac crest and transplanted beneath the latissimus dorsi muscle. Defects of the mandible and oral floor were reconstructed using this combined flap immediately after the resection. The patients began to eat 2 weeks after surgery. [ABSTRACT FROM AUTHOR]

We investigated cellular immunity against Streptococcus pyogenes in human tonsils by measuring antigen-specific immunoglobulin-secreting cells and the production of cytokines from CD4+ T cells in response to M proteins. The incidence of S pyogenes in tonsils was significantly higher in patients with recurrent tonsillitis (RT) than in patients with tonsillar hypertrophy (TH). M protein-specific immunoglobulin A (IgA) and immunoglobulin G spot-forming cells were increased in patients with RT compared with patients with TH. In RT the number of M protein-specific IgA spot-forming cells was significantly greater in the S pyogenes-negative subjects than in the S pyogenes-positive subjects. Proliferation of CD4+ T cells and production of interferon-gamma (IFN-gamma) and interleukins -2, -4, -5, and -6 (IL-2, IL-4, IL-5, and IL-6) from those T cells were observed in response to M protein. The concentrations of IFN-gamma and IL-4 were higher in RT than in TH. These findings suggest that S pyogenes is associated with the pathogenesis of RT and that immune responses against M protein may play an important role in preventing the colonization of this bacteria in tonsils. [ABSTRACT FROM AUTHOR]

Objectives/hypothesis: Intranasal immunization with phosphorylcholine (PC) is known to reduce immunoglobulin (Ig)E production. However, its effects on the occurrence of allergic rhinitis (AR) are unknown. This study was performed to evaluate the effects of PC-keyhole limpet hemocyanin (PC-KLH) and to examine the effects on the occurrence of AR in a murine model of AR., Study Design: In vivo study using an animal model., Methods: Forty-five female BALB/c mice were divided into three groups; those pretreated with intranasal administration of PC-KLH followed by intraperitoneal sensitization and nasal challenge with ovalbumin (OVA) (group A), those untreated with PC-KLH followed by sensitization and nasal challenge with OVA (group B), and those untreated with PC-KLH or OVA as controls (group C). Nasal symptoms, allergic inflammation in the nasal mucosa, OVA specific IgE production, and cytokine profile were compared among those three groups. Dendritic cells (DCs) were isolated from splenic cells and PC-KLH-stimulated interleukin (IL)-12p40 production was measured., Results: The mice pretreated with PC-KLH showed lower allergic nasal symptoms and inflammation compared to untreated mice. The levels of total IgE and OVA-specific IgE in serum, and IL-4 production by nasal and splenic CD4 + T cells were significantly reduced by PC-KLH pretreatment. Furthermore, IL-12p40 production by DCs was induced by PC-KLH in a dose-dependent manner., Conclusions: Intranasal administration of PC-KLH suppressed allergic inflammation in nasal mucosa and antigen-specific IgE production by downregulating Th2-type immune response. Intranasal immunization with PC might be useful to prevent AR and upper airway bacterial infection., Level of Evidence: NA. Laryngoscope, 128:E234-E240, 2018., (© 2017 The American Laryngological, Rhinological and Otological Society, Inc.)

Objective: Pneumococcal infection caused by Streptococcus pneumoniae is a major upper respiratory tract disease that causes severe illness and mortality. Therefore, it is important to develop safe and effective vaccines to prevent pneumococcal infections. The goal of the study was to investigate the effectiveness of transcutaneous immunization (TCI) for induction of pneumococcal surface protein A (PspA) responses in the upper respiratory tract., Methods: C57BL/6 mice were transcutaneously immunized with 1 μg of PspA and 2 μg of cholera toxin (CT) six times at weekly intervals and compared with transcutaneously treated controls (PBS alone/PspA alone/CT alone). Two weeks after the final immunization, nasal washes (NWs), saliva, and plasma samples were collected and subjected to a PspA-specific ELISA. Three weeks after the final immunization, mice were challenged with S. pneumoniae strain EF3030, and the numbers of CFUs in NWs and nasal passages (NPs) were determined., Results: Higher levels of PspA-specific IgM, IgG, and IgA Abs were noted in plasma of TCI with PspA plus CT compared with controls. Transcutaneous immunization mice also had significantly increased PspA-specific S-IgA Ab responses in NWs and saliva and, importantly, showed significantly lower numbers of bacteria CFUs in NWs and NPs compared with controls., Conclusion: These results show that TCI with PspA plus CT induces antigen-specific mucosal and systemic immune responses. This suggests that this method is an effective mucosal immunization strategy for induction of protective pneumococcal-specific Ab responses in blockade of S. pneumoniae colonization of the nasal cavity., Level of Evidence: NA. Laryngoscope, 128:E91-E96, 2018., (© 2017 The American Laryngological, Rhinological and Otological Society, Inc.)

Objectives/hypothesis: Viral upper respiratory tract infections (URIs) are often followed by secondary bacterial infections. To better understand this phenomenon, we examined the impact of the viral agent polyinosine-polycytidylic acid [Poly (I:C)] on the adherence of Streptococcus pneumoniae (Spn) to pharyngeal epithelial cells., Study Design: In vitro model of cultured human pharyngeal epithelial cells., Methods: Detroit 562 cells, a human pharyngeal carcinoma cell line, were pretreated with Poly (I:C). Poly (I:C)-induced expression of platelet-activating factor receptor (PAF-R) was assayed using real-time polymerase chain reaction, flow cytometry, and immunofluorescence microscopy. Bacterial adhesion to these epithelial cells was assessed using immunofluorescence microscopy and colony formation assays., Results: Pretreatment with Poly (I:C) increased mRNA and protein expression of PAF-R in Detroit 562 cells and enhanced the adherence of Spn to these epithelial cells., Conclusions: RNA viral infection can enhance PAF-R expression in epithelial cells and increase the adherence of Spn. These findings might explain in part the mechanisms that underlie the increase in bacterial infection following URIs., (Copyright © 2011 The American Laryngological, Rhinological, and Otological Society, Inc.)

Objectives/hypothesis: One of the major functions of vascular endothelial growth factor (VEGF) is increasing vascular permeability. We previously reported that VEGF is present in nasal secretions in rhinosinusitis, particularly in allergic rhinitis, and is hyperproduced immediately after antigen provocation. However, its function in nasal mucosa has not yet been investigated. This study was designed to estimate the increased vascular permeability by VEGF in nasal mucosa and to discuss its possible role in allergic rhinitis., Study Design: Prospective animal experiment., Methods: Hartley guinea pigs were used. Ethyl carbamate was injected into the abdomen for anesthesia, followed by intravenous injection of Evans blue into the femoral vein. Phosphate buffered saline, histamine, or VEGF was then applied to nasal cavities. Nasal mucosa was harvested after saline reflux from heart with phlebotomy by amputating the abdominal aorta. Tissue samples were soaked in formamide for 12 hours at 63 degrees C to extract dye exudates from nasal mucosa. To analyze vascular permeability, dye was quantified by colorimetry., Results: VEGF increased vascular permeability in nasal mucosa, and was more than 1 x 10(6) times more potent than histamine on the basis of molar units. Although increased vascular permeability by VEGF was not inhibited by histamine 1 receptor antagonist, it was markedly inhibited by anti-VEGF receptor antibodies., Conclusions: VEGF increased vascular permeability to a greater degree than histamine in nasal mucosa via a different pathway. Regulating VEGF may be a new therapeutic option for persistent nasal symptoms in allergic rhinitis.

Objectives: Although otitis media with effusion (OME) is still a common disease in children and adults, the pathogenesis is not yet fully understood. We studied the effects of intratympanic injection with endotoxin purified from nontypeable Haemophilus influenzae on the characteristics of middle ear effusion (MEE)., Methods: Murine model of OME was developed by eustachian tube (ET) blockage followed by intratympanic inoculation with endotoxin (endotoxin group) or saline (control group). The mice were decapitated and histological changes and the production of inflammatory cytokines in MEEs were examined 3 days, 2 weeks, and 2 months after injection., Results: All mice showed OME until 2 months after ET blockage. Most MEEs in the control group were serous, and mucoid or pultaceous MEEs were found only in the endotoxin group. Subepithelial space of middle ear mucosa was severely thickened with the infiltration of a large number of mononuclear cells in the endotoxin group. The levels of tumor necrosis factor-alpha (TNF-alpha) in MEEs were significantly higher in the endotoxin group than in the control group at all time points. Further, in situ hybridization showed that TNF-alpha messenger RNA was expressed not only by leukocytes and macrophages in MEEs but mononuclear cells present in the subepithelial space of middle ear mucosa., Conclusions: These results indicate that ET blockage is essential for the induction of serous MEE and additional administration of endotoxin is associated with the production of mucoid MEE accompanied by histological changes with inflammatory cell infiltration and cytokine production in the tympanic cavity.

Objectives/hypothesis: The production of cytokines by adenoids is known to be associated with inflammation of nasopharynx and the pathogenesis of otitis media with effusion. However, the role of adenoids in producing inflammatory cytokines such as interleukin-8 (IL-8) is not yet clear. In the present study, expression of IL-8 in adenoidal fibroblasts was investigated at the level of transcription factors. Further, the effects of clarithromycin, a 14-member ring macrolide, on IL-8 gene expression and nuclear factor-kappa B (NF-kappa B) activation in adenoidal fibroblasts were evaluated., Study Design: In vitro study for the production of inflammatory cytokine from human adenoidal fibroblasts., Methods: Adenoidal fibroblasts were incubated with nontypeable Haemophilus influenzae endotoxin or interleukin-1 beta. Then the expression of IL-8 and the influence of NF-kappa B inhibitor and clarithromycin were evaluated. Interleukin-8 protein production was assessed by ELISA, and IL-8 messenger RNA production was measured by Northern blot analysis and reverse transcriptase-polymerase chain reaction. Activation of NF-kappa B and inhibition of its activation were determined by electrophoretic mobility shift assay., Results: The expression of both IL-8 protein and messenger RNA in adenoidal fibroblasts was enhanced by Haemophilus influenzae endotoxin and interleukin-1 beta and was positively correlated with increases in NF-kappa B activity. Treatment of cells with the NF-kappa B inhibitor N-tosyl-(L)-phenylalanine chloromethyl ketone, as well as with clarithromycin, reduced expression of IL-8 and NF-kappa B activity in a dose-dependent manner., Conclusions: Results suggest that adenoidal fibroblasts produce IL-8 in response to endotoxin through NF-kappa B activation. The inhibitory effects of clarithromycin on NF-kappa B activation and IL-8 production in adenoidal fibroblasts might explain, in part, the mechanism of this drug in improving otitis media with effusion.

BALB/c mice were immunized orally or subcutaneously with formalin-killed nontypeable Haemophilus influenzae (NTHi). Salivary immunoglobulin A (IgA) antibody titers against NTHi were significantly increased by oral immunization, but not by subcutaneous immunization. Both immunization procedures remarkably increased the levels of serum antibody activities of both IgA and immunoglobulin G. Live NTHi were then inoculated into the naso-pharynx, and the clearance of the pathogen from the nasopharynx was observed. Significantly fewer bacteria were present in the nasopharynx of the orally immunized mice than in the control mice. However, there was no significant difference between the subcutaneously immunized mice and the control mice. The results indicate that oral immunization can enhance the ability of mice to clear NTHi from the nasopharynx.

Reflectance spectrophotometry analysis has been applied to the human nasal mucosa in vivo to determine the objective parameters of the colors of the mucous membrane, as well as to study the hemodynamics and enzymatic activities related to respiratory chain metabolism in the nasal mucosa. In 24 normal subjects, there were three main spectral peaks: the peak at around 500 nm (PI), the smallest at 560 nm (P2) and the highest one at 660 nm (P3). In 28 patients with nasal allergy, spectral P3 shifted remarkably toward the shorter wavelength at around 640 nm, compared to that of normal subjects. Additionally, the increment of the absorption band at P2 was also observed. These phenomena could be seen in the nasal mucosa of the subjects after use of decongestant spray. Reflectance spectrophotometric analysis will serve as a non-invasive method of giving us useful information for a further understanding of the mucosal pathophysiology of the upper airway.