Fibrinolytic components in nasal mucosa and nasal secretion

 We evaluated a possible role for fibrinolytic components in nasal secretion by tissue localization with immunohistochemical techniques and by measuring their antigen concentrations in nasal discharge by means of ELISA and fibrin autography. Nasal mucosa was obtained surgically from the inferior turbinate. Urokinase-type plasminogen activator (u-PA) specific staining was observed in pseudostratified ciliated epithelium and was predominant in mucous cells of the seromucinous gland, while serous cells were almost devoid of stain. The pattern of staining of plasminogen activator inhibitor-2 was similar to that of u-PA. In contrast, plasminogen activator inhibitor-1(PAI-1) immunoreactive material was localized exclusively in serous cells of seromucinous glands. Positive staining for tissue-type plasminogen activator (t-PA) was observed in endothelial cells and basal cells, which differentiate into either ciliated or goblet cells. Nasal secretions were partially fractionated by immunospecific antibody-immobilized Sepharose. Subsequent fibrin autography patterns indicated the presence of u-PA, PAI-1, and t-PA. After methacholine provocation, the level of t-PA increased transiently but decreased rapidly with subsequent challenges. These differential stainings of fibrinolytic components and the existence of PAs and PAI-1 in the nasal discharge suggest that the fibrinolytic system may play a role in the movement and fluidity of nasal secretion. Accepted: 25 May 1998     

Microbiocenosis of nasal mucous membrane during allergic rhinosinusitis

Overall quantity of microorganisms and their relative prevalence on nasal mucous membrane in patients with allergic rhinosinusitis are determined. Microbial flora in studied group was characterized qualitatively and quantitatively. Obtained results showed that there were changes of microbiocenosis of nasal mucous membrane during allergic rhinosinusitis and in characteristics of species in Staphylococcus genus.     

A quantitative analysis of cedar pollen-specific immunoglobulins in nasal lavage supported the local production of specific IgE, not of specific IgG

Many studies have proved the relevance of local immune responses, rather than systemic immunity, to the pathogenesis of allergic rhinitis. Indeed, allergen-specific B lymphocyte undergoes class switching to IgE in situ. However, the relative contribution of in situ production to the amount in serum is still ambiguous. Here, a quantitative comparison of the local concentration of allergen-specific IgE with the systemic concentration was explored for the estimation. Among seasonal rhinitis patients, total and Japanese cedar pollen (JCP)-specific IgE, IgA and IgG antibodies were quantified in nasal lavage fluid (NLF) and serum with the time-resolved fluorescence immunosorbent assay. Although the total amounts of IgE and IgG classes in the NLF, which were apparently passive discharge from the mucosal tissue, were smaller and variable, the relative proportions of JCP-specific antibodies could be quantitatively compared between NLF and serum or between subjects. The proportions of specific IgE in the NLF were remarkably higher than in serum (average 13.2-fold) in most subjects, which strongly supported the predominant in situ production of the specific IgE and subsequent dilutions in the systemic circulations. Similar but smaller values were obtained for IgA (average 3.7-fold). In contrast, the specific proportions of IgG in the NLF were surprisingly consistent with serum (average 1.0-fold), suggesting that the specific IgG was mostly produced in the downstream lymphoid organs. The local productions of specific IgE would encourage the topical therapies and the usage of the NLF for the diagnosis of allergic rhinitis.     

Histochemical study of glycoconjugates in the nasal mucosa of the rat and guinea pig

Summary A histochemical study was carried out on the glycoconjugates of the nasal mucosa of rat and guinea pig using conventional techniques and peroxidase-labelled lectins. Both the respiratory mucosa and neuroepithelium were studied. Sulphate and sialic acid groups were found in the mucous layer of the neuroepithelia, Bowman's glands and goblet cells. In contrast, the nasal glands did not possess these groups, and only a few showed neutral mucins. Carbohydrate residues were more numerous in the acini of the Jacobson glands. Thus, the nasal glands in the rat and guinea pig are probably of a serous type because of the scarcity of carbohydrate residues.     

Neuroendocrine cells in the nasal mucosa - preliminary report

The role of neuroendocrine cells (NC) in physiology and pathology of the human's respiratory tract is not fully understood. The aim of the study was the quantitative and morphometric assessment of NC in nasal mucosa in some pathological states. PATIENTS AND METHOD: 40 patients, aged 28-63 years, with clinical signs of chronic, hypertrophic rhinosinusitis were qualified for the study. Rhinitis chronica hypertrophica coexisted with aspirin triad or asthma in 10 patients (group I), with advanced obstructive sleep apnea syndrome (OSAS) in 10 patients (respiratory disturbance index, RDI>40, group II). Group III consisted of 10 patients with simple rhinitis chronica hypertrophica who habitually smoked cigarettes (at least 20 cigarettes a day) while 10 non-smoking patients with simple rhinitis chronica hypertrophica were qualified to the control group. Fragments of nasal mucosa of approximately 0.5 cm(2) were collected from medial or inferior turbinate during mucoplasty procedures. NC were detected immunohistochemically using antibodies against chromogranin A (DAKO). The microscopic sections were evaluated in the light microscopy. RESULTS: The study did not reveal the increased number of NC in examined fragments of nasal mucosa. Scattered NC were detected in single preparations of nasal mucous membrane in some patients in all groups. The number of detected neuroendocrine cells did not differ statistically between groups.     

Polarized cultures of human airway epithelium from nasal scrapings and bronchial brushings

Airway epithelial cultures are generally derived from tracheas postmortem or from surgical specimens of nasal polyps or turbinates. Scrapings of the mucosal surface have been little used as starting material for cultures because of their low yield of epithelial cells and their contamination with mucous secretions, blood, and underlying connective tissue. For the first time, we report that human airway epithelial cells obtained from nasal scrapings or bronchial brushings can be grown in culture to produce polarized cell sheets suitable for studies of vectorial transport.     

Mucosal adjuvant activity of oligomannose-coated liposomes for nasal immunization

In the present study, we investigated the effectiveness of liposomes coated with a neoglycolipid consisting of mannotriose and dipalmitoylphosphatidylcholine (Man3-DPPE) as an adjuvant for induction of mucosal immunity. Immunization of BALB/c mice with ovalbumin (OVA)-encapsulated Man3-DPPE-coated liposomes (oligomannose-coated liposomes; OMLs) by a nasal route produced high levels of OVA-specific IgG and IgA antibodies in serum of immunized mice 1 week after the last nasal immunization, whereas no significant serum antibody responses were observed in mice that received OVA in uncoated liposomes or OVA alone. Seven weeks after the last nasal immunization, nasal challenge with an excess amount of OVA in mice that had received OVA/OMLs led to an anamnestic response to the antigen that resulted in 5- to 10-fold increases of antigen-specific serum IgG and IgA antibodies. Only mice immunized nasally with OML/OVA secreted antigen-specific secretory IgA in nasal washes and produced interferon-gamma secreting cells in nasopharyngeal-associated lymphoreticular tissue. Taken together, these results show that nasal administration of OMLs induces mucosal and systemic immunity that are specific for the entrapped antigen in the liposomes. Thus, liposomes coated with synthetic neoglycolipids might be useful as adjuvants for induction of mucosal immunity.     

Expression and localization of the thromboxane A2 receptor in human nasal mucosa

Thromboxane A2 (TXA2) has been thought a potent mediator involved in allergic rhinitis, because TXA2 was recovered from the nasal lavage fluid of allergic rhinitis patients after allergen provocation and TXA2 receptor antagonists relief nasal allergic symptoms. In order to clarify the expression of TXA2 receptor in human nasal mucosa, we investigated TXA2 receptor mRNA expression and its protein localization by polymerase chain reaction (PCR) and immunohistochemistry, respectively. Human turbinates were obtained after turbinectomy from 10 patients with nasal obstruction refractory to medical therapy. RT-PCR analysis of total RNA from nasal mucosa demonstrated the expression of TXA2 receptor alpha mRNA. The immunohistochemical studies revealed that anti-TXA2 receptor alpha antibody labeled vascular smooth muscle cells, vascular endothelial cells, epithelial cells and submucosal glands in the nasal mucosa. The results may have an important clinical implication for understanding the role of TXA2 receptor on upper airway diseases such as allergic rhinitis and non-allergic rhinitis.     

Participation of complement in nasal polyposis

No changes in immunoglobulin (IgG, IgM and IgA) and complement components C3 and C4 levels could be observed in patients suffering from both recurrent and acute nasal polyposis. However, in both groups of patients a significant decrease of the alternative pathway of complement activation (up to 80%) was found. In addition, patients suffering from the acute disease showed even decreased classical complement pathway (up to 20%). Examination of complement activation pathways was shown to be advantageous for the follow-up of the course of this disease.     

Frequency of specific anti-Toxoplasma gondii IgM, IgA and IgE in colombian patients with acute and chronic ocular toxoplasmosis

We studied the frequency of specific anti-Toxoplasma IgM, IgA and IgE antibodies in serum of 28 immunocompetent Colombian patients, selected by ophthalmologists and with lesions that were compatible with ocular toxoplasmosis. Patients were classified in three groups: (i) group 1 consisted of ten patients with a first episode; (ii) group 2, with seven patients with a recurrence and (iii) group 3, consisted of eleven patients with chronic chorioretinal lesion without uveitis. We found that 10/28 (35%) of Colombian patients with ocular toxoplasmosis possessed at least one serological marker for Toxoplasma infection different from IgG. In group 1 (first episode), we found simultaneous presence of specific IgM plus IgA plus IgE in 1/10 (10%). In group 2 (recurrences) in 1/7 (14%) we found IgM and IgA test positives and in 1/7 (14%) we found IgM and IgE tests positives. In group 3 (toxoplasmic chorioretinal scar) the IgA serological test was positive in 2/11 (18%). These results show that serum IgM or IgA or IgE can be present during recurrences.     

An immunohistochemical study of cells with surface and cytoplasmic immunoglobulins in situ in Peyer's patches and lamina propria of rat small intestine

The distribution of cells with surface and cytoplasmic immunoglobulins was studied in Peyer's patches (PP) and intestine of rats, using both frozen and paraffin sections, with a two-step peroxidase technique. Anti IgM, IgG, IgA and IgE sera were used. Surface staining was found within PP with all antisera used. Although the villi contained predominantly IgA plasma cells (PC), IgG PC and a few IgM and IgE PC were also found. Within PP, however, no IgA PC were found but IgM and IgG PC were present in all stages of development, mainly in the dome. PC of all types, but mostly IgA cells, were present in and around high endothelial venules (HEV). The results suggest that IgM and IgG PC precursors can develop to PC within PP whereas IgA precursors do not. PC appear to home to the gut preferentially via HEV.     

小鼠胚胎发育过程中表皮生长因子受体的免疫荧光定位研究

用间接免疫荧光法研究了第12—19天小鼠胚胎的表皮生长因子受体(EGFR)在各种组织中的分布。结果表明,特异性荧光出现在EGFR阳性细胞的细胞膜上。第14.5天鼠胚鼻粘膜上皮首先显示很强的EGFR特异性荧光,此后荧光稍为减弱,直至第19天后消失。消化系统中,舌味觉上皮在第16天、胃粘膜上皮在第17—18天间、十二指肠上皮在第17.5—18.5天、直肠粘膜上皮在第15.5—16天和肛管粘膜上皮在第15天均显示强特异性荧光。肝细胞从第14.5天起有弱阳性反应,随胎龄增大强度缓慢地增强。此外,在胚胎发育不同时期,还看到若干组织呈现阳性反应,包括膀胱粘膜变移上皮、眼睑原基、腺垂体上皮、舌下腺腺泡细胞、附属腺上皮细胞、胰岛细胞、颌下腺腺上皮和导管上皮细胞、降主动脉内皮以及卵巢髓质部富含血管的疏松结缔组织中的成纤维细胞等。     

A roentgenographic method for measuring nasal mucous velocity

Nasal mucous velocity was estimated by following the motion of radiopaque discs of Teflon by means of a fluoroscopic image intensifier. From 5 to 10 discs were deposited on the superior surface of the inferior turbinate with a forceps. No local anesthesia was employed and the subjects experienced no discomfort. The linear velocity of the discs was obtained by playing the videotape onto a television monitor, measuring distance with a ruler, and dividing by elapsed time. Duplicate runs of 1-2 min, 15 min apart were very reproducible but runs at 4-h intervals or daily over a 5-day period had a coefficient of variation of 30%. Average nasal velocity for individual ranged from 0 to 22.5 mm/min and group means ranged from 6. 8 to 10.8 mm/min. There was no statistically significant difference in nasal mucous velocity between young and elderly subjects nor was there a sexual difference. The saccharin test of nasal mucous transport was unsatisfactory because of inability to repeat the test more often than 1-2 h and its propensity to produce mild discomfort in a significant number of subjects. Saccharin times did not correlate significantly with values of nasal mucous velocity.     

Application of chitosan microspheres for nasal delivery of vaccines

Nasal vaccines offer several benefits, such as highly vascular mucous membranes, low enzymatic degradation compared to oral vaccines, and greater acceptability to patients. Nasal vaccines, however, have to overcome several limitations, including mucociliary clearance and the inefficient uptake of soluble antigens. Therefore, nasal vaccines require potent adjuvants and delivery systems to enhance their immunogenicity and to protect their antigens. Chitosan is a cheap, biocompatible, biodegradable, mucoadhesive, and nontoxic natural polymer. Chitosan microspheres have been investigated to determine whether they allow the controlled release of drugs and vaccines. They have figured in various studies on the vaccine delivery system through the nasal route. Several researchers have developed modified chitosan microspheres through their concomitant use with adjuvants or immunomodulators for an additive and a synergistic effect, and through the mannosylation of chitosan for receptor-mediated targeting antigen-presenting cells. The results of the recent researches on chitosan microspheres used as a nasal vaccine delivery system are discussed in this review.     

Serum immunoglobulins in patients with carcinoma of the oral cavity,uterine cervix and breast

Summary Serum immunoglobulin levels (IgG, IgA, IgM, IgD and IgE) were estimated in 196 patients with carcinoma of the oral cavity, 172 patients with cervical cancer and 166 patients with breast cancer. The values were compared with those of 50 patients with benign lesions of the breast and cervix and 100 healthy adult controls. Only the serum IgE levels were found to be elevated in the benign group. Serum IgA, IgD and IgE levels were found to be elevated in all the three types of cancers and the levels were found to increase with clinical stage. In carcinoma of uterine cervix IgG levels were also found to be elevated. Immunoglobulins A and D returned to normal after clinical cure whereas IgE remained slightly elevated. IgD and IgE remained high in patients who had residual cancer.     

Combined oral/nasal immunization protects mice from Sendai virus infection

Based on the concept of a common mucosal immune system wherein mucosal associated lymphocytes traffic among the various mucous membranes, the murine gastrointestinal tract was immunized with Sendai virus antigens in order to elicit a virus-specific immune response in the respiratory tract. Multiple intragastric (oral) administration of live or killed Sendai virus induced IgA and IgG antiviral antibodies in both gastrointestinal secretions and serum. When cholera toxin as an adjuvant was included along with virus, gut IgA and IgG as well as serum IgA responses were enhanced. Antiviral antibodies induced in respiratory secretions by oral killed virus plus cholera toxin, however, were variable and protection from virus challenge was not demonstrated. Significantly higher levels of respiratory antiviral antibodies were induced if immunization with oral killed Sendai virus/cholera toxin was combined with intranasal administration of small amounts of killed virus. The combined immunization also resulted in protection of both the upper and lower respiratory tracts from virus infection. Protection of the upper respiratory tract was correlated with the presence of IgA antiviral antibodies in nasal washings. On the other hand, protection of the lower respiratory tract was correlated with IgG antiviral antibodies in bronchoalveolar lavage fluids. Immunization with intranasal killed virus alone conferred partial protection to the lower respiratory tract and no protection to the upper respiratory tract. Thus, oral immunization with killed virus antigen could prime for a protective immune response in the murine respiratory tract and this protective response included IgA antibodies.     

Localization of the human neonatal Fc receptor (FcRn) in human nasal epithelium

The airway epithelium is a central player in the defense against pathogens including efficient mucociliary clearance and secretion of immunoglobulins, mainly polymeric IgA, but also IgG. Pulmonary administration of therapeutic antibodies on one hand, and intranasal immunization on the other, are powerful tools to treat airway infections. In either case, the airway epithelium is the primary site of antibody transfer. In various epithelia, bi-polar transcytosis of IgG and IgG immune complexes is mediated by the human neonatal Fc receptor, FcRn, but FcRn expression in the nasal epithelium had not been demonstrated, so far. We prepared affinity-purified antibodies against FcRn α-chain and confirmed their specificity by Western blotting and immunofluorescence microscopy. These antibodies were used to study the localization of FcRn α-chain in fixed nasal tissue. We here demonstrate for the first time that ciliated epithelial cells, basal cells, gland cells, and endothelial cells in the underlying connective tissue express the receptor. A predominant basolateral steady state distribution of the receptor was observed in ciliated epithelial as well as in gland cells. Co-localization of FcRn α-chain with IgG or with early sorting endosomes (EEA1-positive) but not with late endosomes/lysosomes (LAMP-2-positive) in ciliated cells was observed. This is indicative for the presence of the receptor in the recycling/transcytotic pathway but not in compartments involved in lysosomal degradation supporting the role of FcRn in IgG transcytosis in the nasal epithelium.     

Distribution of human PLUNC/BPI fold-containing (BPIF) proteins

Although gene expression studies have shown that human PLUNC (palate, lung and nasal epithelium clone) proteins are predominantly expressed in the upper airways, nose and mouth, and proteomic studies have indicated they are secreted into airway and nasal lining fluids and saliva, there is currently little information concerning the localization of human PLUNC proteins. Our studies have focused on the localization of three members of this protein family, namely SPLUNC1 (short PLUNC1), SPLUNC2 and LPLUNC1 (long PLUNC1). Western blotting has indicated that PLUNC proteins are highly glycosylated, whereas immunohistochemical analysis demonstrated distinct patterns of expression. For example, SPLUNC2 is expressed in serous cells of the major salivary glands and in minor mucosal glands, whereas SPLUNC1 is expressed in the mucous cells of these glands. LPLUNC1 is a product of a population of goblet cells in the airway epithelium and nasal passages and expressed in airway submucosal glands and minor glands of the oral and nasal cavities. SPLUNC1 is also found in the epithelium of the upper airways and nasal passages and in airway submucosal glands, but is not co-expressed with LPLUNC1. We suggest that this differential expression may be reflected in the function of individual PLUNC proteins.