lactoferrin and Sinusitis

Il profilo di espressione genica multipla rivela una disfunzione epiteliale nella rinosinusite cronica polipoide.. La rinosinusite cronica (CRS) è un disturbo infiammatorio eterogeneo risultante da una complessa interazione genetico-ambientale. Sebbene l’eziologia rimanga tuttora sfuggente, numerosi studi riportano alterazioni nell’espressione genica di diversi fattori implicati nell’ambito della risposta infiammatoria. Tuttavia, la gran parte di queste sono analisi isolate, non replicate, che prendono in considerazione un singolo gene alla volta. Inoltre, gli studi riguardanti analisi di espressione genica multipla, solitamente su mediatori infiammatori (es. citochine), spesso presentano risultati contrastanti, che in parte possono essere dovuti all’eterogeneità dei campioni o a metodologie analitiche di potenza limitata. In quest’ottica, il nostro obiettivo è stato di verificare simultaneamente l’espressione genica di un pannello di geni (AQP5, MUC5AC, CAV1, LTF, COX2, PGDS, TNFα, TGFβ1, MGB1) potenzialmente coinvolti nei meccanismi infiammatori della CRS. Nonostante la gran parte dei campioni sia stata esclusa dall’analisi a causa del deterioramento dell’RNA tissutale, siamo stati in grado di dimostrare una riduzione statisticamente significativa dell’espressione dei geni AQP5, CAV1, LTF e MGB1, in uno specifico sottogruppo di pazienti affetti da CRS nella variante con polipi nasali senza le tipiche comorbidità frequentemente associate (asma, allergia, intolleranza all’acido acetil-salicilico). Questi dati sembrano suggerire una disfunzione della barriera epitaliale nella CRS polipoide. Ulteriori studi saranno necessari per incrementare ulteriormente la nostra conoscenza sulla patogenesi della CRS. A tal proposito l’applicazione delle nuove e più potenti tecniche di sequenziamento, come la next-generation RNA sequencing, e la disponibilità di analisi bioinformatiche più complete potranno migliorare la caratterizzazione del transcriptoma negli endotipi della CRS.. Chronic rhinosinusitis (CRS) is a heterogeneous inflammatory disorder resulting from a complex gene-environment interaction. Although its aetiology remains elusive, numerous studies reported gene expression alterations of factors apparently implicated in all aspects of the inflammatory response. However, most investigations are limited, unconfirmed analyses of a single gene. Moreover, studies concerning multiple gene expression analyses, usually on inflammatory mediators (e.g. cytokines), show contrasting outcomes in part due to use of heterogeneous samples or methodologies with limited power. In this scenario, our goal was to simultaneously evaluate the expression of a panel of selected genes (AQP5, MUC5AC, CAV1, LTF, COX2, PGDS, TNFα, TGFβ1, MGB1) potentially involved in CRS inflammatory mechanisms. While most of the samples collected were excluded from the analysis because of poor quality RNA, we were able to demonstrate statistically significant downregulation of the AQP5, CAV1, LTF, MGB1 genes in a specific subset of polypoid CRS (patients without typical comorbidities), which might suggest relevant underlying epithelial dysfunction. Further studies are needed to enrich our knowledge on the pathogenesis of CRS. Forthcoming approaches might utilise next-generation RNA sequencing and comprehensive bioinformatics analyses to better characterise the transcriptome profiles of CRS endotypes.

Topics: Adolescent; Adult; Aged; Aquaporin 5; Caveolin 1; Chronic Disease; Cyclooxygenase 2; Cytokines; Epithelial Cells; Gene Expression; Gene Expression Profiling; Humans; Lactoferrin; Mammaglobin A; Middle Aged; Mucin 5AC; Nasal Polyps; Retrospective Studies; Rhinitis; Sinusitis; Transforming Growth Factor beta1; Tumor Necrosis Factor-alpha; Young Adult

Both up- and down-regulation of the Toll-like receptors (TLRs) and antimicrobial peptides (AMPs) of the sinonasal mucosa have already been associated with the pathogenesis of chronic rhinosinusitis with (CRSwNP) or without (CRSsNP) nasal polyps. The objective of this study was to determine the expression of all known TLR and several AMP genes and some selected proteins in association with allergy, asthma and aspirin intolerance (ASA) in CRS subgroups. RT-PCR was applied to measure the mRNA expressions of 10 TLRs, four defensins, lysozyme, cathelicidin and lactoferrin (LTF) in sinonasal samples from patients with CRSsNP (n = 19), CRSwNP [ASA(-): 17; ASA(+): 7] and in control subjects (n = 12). Protein expressions were detected with immunohistochemistry (n = 10). Statistical analysis was done with the Kruskal-Wallis ANOVA, Mann-Whitney U, and Student t test. TLR2, TLR5, TLR6, TLR7, TLR8, TLR9, β-defensins 1 and 4, cathelicidin and LTF mRNA expressions were significantly (p < 0.05) increased in CRSwNP, whereas only TLR2 and LTF were up-regulated in CRSsNP compared to controls. There was no statistical difference in respect of allergy, aspirin intolerance and smoking between CRSsNP, ASA(-) and ASA(+) CRSwNP patients. TLR2, TLR3, TLR4, LTF, β defensin 2 and lysozyme protein expressions were found to be elevated in macrophages of CRSwNP samples (p < 0.05). Gene expression analysis showed markedly different expressions in CRSwNP (6 out of 10 TLR and 4 out of 7 AMP genes were up-regulated) compared to CRSsNP (1/10, 1/7). The distinct activation of the innate immunity may support the concept that CRSsNP and CRSwNP are different subtypes of CRS. These findings were found to be independent from allergy, asthma, smoking, aspirin intolerance and systemic steroid application.

Topics: Adult; Antimicrobial Cationic Peptides; beta-Defensins; Case-Control Studies; Cathelicidins; Chronic Disease; Female; Humans; Hypersensitivity; Lactoferrin; Male; Middle Aged; Nasal Polyps; Rhinitis; RNA, Messenger; Sinusitis; Toll-Like Receptors; Young Adult

The objective of the present work was to study the contribution of biofilms to the development of chronic bacterial rhinosinusitis. A total of 50 patients with this pathology were available for the examination. Mucosal swabs were taken from the middle nasal passages of all the patients to be used for the detection of biofilms by luminescence microscopy. The lactoferrine content in mucosal secretion from the nasal cavity was determined by an immunoenzymatic assay. Two groups of the patients presenting with bacterial rhinosinusitis were distinguished in the study of impression smears from nasal cavity mucosa by luminescence microscopy; one of them was comprised of biofilm-positive patients the other of biofilm-negative ones (56% and 44% respectively). The patients showing biofilms over nasal cavity mucosa had the lactoferrine content in mucosal secretion on the order of 0.0033±0.0008 mg/l compared with 0.0068±0.00075 mg/l in the biofilm-negative patients and 0.55±0.0005 mg/l in the healthy volunteers (controls). In other words, the biofilm-positive patients presenting with chronic bacterial rhinosinusitis had two times lower content of lactoferrine in mucosal secretion from the middle nasal passages than those in the biofilm negative group and 126 times lower content of lactoferrine than in the control group.

Topics: Adolescent; Adult; Bacterial Infections; Biofilms; Chronic Disease; Female; Humans; Lactoferrin; Male; Middle Aged; Nasal Mucosa; Rhinitis; Sinusitis; Young Adult

Nasal polyps are strongly associated with a risk of chronic rhinosinusitis development as well as other obstruction including asthma and allergy. The following study tested the association of the 140A/G polymorphism of lactoferine (LF) encoding gene and the -33C/G polymorphism of osteoblast-specific factor-2 (OSF-2) encoding gene with a risk of chronic rhinosinusitis with nasal polyps in a Polish population. One hundred ninety five patients of chronic rhinosinusitis with nasal polyps as well as 200 sex, age and ethnicity matched control subjects without chronic sinusitis and nasal polyps were enrolled in this study. Among the group of patients 63 subjects were diagnosed with allergy and 65 subjects with asthma, respectively. DNA was isolated from peripheral blood lymphocytes of patients as well as controls and gene polymorphisms were analyzed by restriction fragments length polymorphism polymerase chain reaction (RFLP-PCR). We reported that the 140A/G LF (OR 4.78; 95% CI 3.07-7.24), the -33C/G OSF-2 OR 3.48; 95% CI 2.19-5.52) and the -33G/G OSF-2 (OR 16.45; 95% CI 6.71-40.30) genotypes were associated with an increased risk of chronic rhinosinusitis with nasal polyps among analyzed group of patients. Moreover, the group of patients without allergy or asthma indicated the association of the -33C/G (OR 3.72; 95% CI 2.24-6.19 and OR 15.11; 95% CI 5.91-38.6) and -33G/G (OR 3.73; 95% CI 2.24-6.19 and OR 14.07; 95% CI 5.47-36.16) genotypes of the OSF-2 as wells as 140A/G (OR 3.89; 95% CI 2.40-6.31 and OR 3.62; 95% CI 2.45-5.34) genotype of OSF-2 with an increased risk of chronic rhinosinusitis with nasal polyps. Finally, it was also found that the selected group of patients with allergy or asthma indicated a very strong association of the -33C/G (OR 2.40; 95% CI 1.23-4.69 and OR 2.40; 95% CI 1.23-4.69, respectively) and -33G/G (OR 16.01; 95% CI 5.77-44.41 and OR 17.90; 95% CI 6.53-49.05, respectively) genotypes of the OSF-2 as wells as 140A/G (OR 3.22; 95% CI 1.74-6.11 and OR 3.25; 95% CI 1.75-6.04, respectively) genotypes with an increased risk of chronic rhinosinusitis with nasal polyps. Thus, our results suggest that LF and OSF-2 gene polymorphisms may have deep impact on the risk of rhinosinusitis nasal polyps' formation which may also depend on asthma or allergy. Our results showed that the 140A/G polymorphism of LF gene and the -33C/G polymorphism of the OSF-2 gene may be associated with the risk of chronic rhinosinusitis with nasal polyps in a Polish pop

Topics: Adult; Aged; Alleles; Case-Control Studies; Cell Adhesion Molecules; Chronic Disease; Confidence Intervals; Electrophoresis, Agar Gel; Female; Gene Frequency; Genetic Association Studies; Genetic Predisposition to Disease; Humans; Lactoferrin; Male; Middle Aged; Nasal Polyps; Odds Ratio; Poland; Polymerase Chain Reaction; Polymorphism, Restriction Fragment Length; Polymorphism, Single Nucleotide; Risk Factors; Sinusitis

Chronic rhinosinusitis (CRS) is a disease characterized by inflammation of the nasal mucosa and paranasal sinuses. This inflammation may result in part from decreased epithelial barrier and innate immune responses, leading to frequent bacterial and fungal colonization. The objectives of this study were to investigate the expression of innate immune proteins of the palate lung and nasal epithelium clone (PLUNC) family in patients with CRS.. Nasal tissue samples were collected from control subjects and CRS patients with and without nasal polyps. Expression of the members of the PLUNC family was analyzed by real-time PCR. Expression of SPLUNC1 and LPLUNC2 proteins was analyzed by ELISA, immunoblot, and immunohistochemical analysis.. Levels of mRNA for most of the members of the PLUNC family were profoundly reduced in nasal polyps (NPs) compared to uncinate tissue from control subjects or patients with CRS. LPLUNC2 and SPLUNC1 proteins were decreased in NPs of patients with CRS compared to uncinate tissue from control subjects. Immunohistochemical data revealed that within submucosal glands of sinonasal tissues, SPLUNC1 and LPLUNC2 were differentially expressed, in serous and mucous cells, respectively. The decrease in the expression of these molecules is probably explained by a decrease in the number of glands in NPs as revealed by correlations with levels of the glandular marker lactoferrin.. Decreased SPLUNC1 and LPLUNC2 in NPs reflect a profound decrease in the number of submucosal glands. Decreased glands may lead to a localized defect in the production and release of glandular innate defense molecules.

Topics: Adolescent; Adult; Aged; Chronic Disease; Female; Gene Expression; Gene Expression Profiling; Gene Expression Regulation; Glycoproteins; Humans; Lactoferrin; Male; Middle Aged; Nasal Mucosa; Nasal Polyps; Phosphoproteins; Rhinitis; Sinusitis; Young Adult

The diverse antipathogenic action of lactoferrin has been well characterized. In addition, it is the human body's only known antimicrobial peptide with antibiofilm properties. The purpose of this study was to examine the nasal mucosal expression of lactoferrin in the biofilm-mediated disease, chronic rhinosinusitis (CRS).. Nasal biopsies from 41 CRS patients and 21 healthy controls were analyzed using confocal scanning laser microscopy (CSLM) and scanning electron microscopy (SEM) for the presence of biofilms. The messenger ribonucleic acid (mRNA) and protein level of lactoferrin in this tissue were also determined by quantitative real-time reverse-transcriptase polymerase chain reaction (qRT-PCR) and enzyme linked immunosorbent assay (ELISA), respectively.. Lactoferrin expression in chronic rhinosinusitis patients at both mRNA and protein level was downregulated relative to controls. Biofilm-positive CRS patients showed a much greater reduction in lactoferrin expression than biofilm-negative patients; mRNA median fold change biofilm positive = 0.03 (interquartile range 0.005-0.15) and biofilm-negative CRS median fold change = 0.49 (interquartile range 0.15-0.81) with median lactoferrin protein expression biofilm-positive patients' median lactoferrin protein expression = 32.58 ng/mL (interquartile range 8.67-59.9 ng/mL) and biofilm-negative patients' median lactoferrin expression = 114.40 ng/mL (interquartile range 75.41-163.1 ng/mL).. Genetic, transcriptional, or translational deficiencies in lactoferrin synthesis may reduce the functional level of this important antimicrobial/antibiofilm peptide in the nasal secretions of CRS patients, predisposing certain individuals to bacterial colonization, biofilm development, and recalcitrant sinus disease.

Topics: Biofilms; Chronic Disease; DNA Primers; Enzyme-Linked Immunosorbent Assay; Gene Expression; Humans; Lactoferrin; Microscopy, Electron; Nasal Mucosa; Prospective Studies; Reverse Transcriptase Polymerase Chain Reaction; RNA; RNA, Messenger; Sinusitis

Antimicrobial peptides, such as lactoferrin, are an important component of the innate immune system. They offer the body a first line defense against a wide range of invading pathogens. The diverse antipathogenic action of lactoferrin has been well characterized; however, the role that this peptide plays in chronic conditions such as rhinosinusitis remains largely unknown. This study aims to examine the level of lactoferrin expression in the nasal mucosa of patients with chronic rhinosinusitis (CRS).. Nasal biopsies of 85 chronic rhinosinusitis patients, subclassified into allergic fungal sinusitis (AFS), nonallergic fungal eosinophilic sinusitis (NAFES), nonallergic, nonfungal eosinophilic sinusitis (NANFES), and CRS were studied by quantitative real-time reverse-transcriptase polymerase chain reaction and enzyme-linked immunosorbent assay for their expression of lactoferrin at an mRNA and protein level, respectively.. All groups of patients with CRS showed a decrease in lactoferrin mRNA expression relative to controls (median fold-change of CRS relative to controls, 0.1550; AFS, 0.1800; NANFES, 0.1900; and NAFES, 0.2100). All groups also showed a decreased expression of lactoferrin protein (controls, 163.3 ng/mL; CRS, 82.19 ng/mL; AFS, 104.1 ng/mL; NANFES, 118.9 ng/mL; and NAFES, 74.33 ng/mL). The most significant reduction was evident in the CRS subgroup as well as in patients with nasal polyposis at the time of surgery.. This is the first study of its kind to objectively examine lactoferrin expression in the nasal mucosa of CRS patients. We report a reduction in the expression of this important antimicrobial peptide at both the mRNA and protein level. Such a defect in the innate immune system may explain the predisposition of certain individuals to develop CRS and nasal polyposis, providing further insight into the pathogenesis of such conditions.

Topics: Adult; Aged; Aged, 80 and over; Chronic Disease; Enzyme-Linked Immunosorbent Assay; Female; Humans; Lactoferrin; Male; Middle Aged; Nasal Mucosa; Reverse Transcriptase Polymerase Chain Reaction; Rhinitis; RNA, Messenger; Sinusitis; Statistics, Nonparametric

The association of perennial allergic rhinitis (PAR) with recurrent sinusitis (RS) is well recognized. Anatomic abnormalities at the osteomeatal complex or ciliary dysfunction may play a significant role in some patients. However, for most patients with allergy, the determinants of RS are unknown.. To determine whether altered concentrations of antimicrobial peptides and proteins, such as lysozyme, lactoferrin, human beta-defensin-2 (HBD-2), and human neutrophil peptides 1 to 3 (HNP-1 to 3), contribute to the development of RS in patients with PAR.. Nasal secretions were collected by vacuum aspiration from 15 individuals with PAR+RS, 16 with PAR alone, and 16 controls. Lysozyme and lactoferrin levels were determined in nasal secretions by using quantitative enzyme-linked immunosorbent assay, and HBD-2 and HNP-1 to 3 levels were determined in nasal secretions by using semiquantitative Western blot analysis. Eosinophil-derived neurotoxin (EDN) levels were measured by using enzyme-linked immunosorbent assay as a marker of nasal eosinophilia in all 3 groups.. Levels of EDN were elevated significantly in patients with PAR+RS compared with controls. Lysozyme levels were decreased significantly in patients with PAR+RS compared with PAR alone or controls. Mean lysozyme levels were significantly lower in patients with EDN levels greater than 1,000 ng/mL vs those with levels of 1,000 ng/mL or less in the PAR+RS group. There were no statistically significant differences in lactoferrin, HBD-2, and HNP-1 to 3 levels among the 3 groups.. The presence of eosinophils and their products and reduced lysozyme concentrations may be critical factors that predispose the airways of patients with PAR to RS.

Topics: Adult; alpha-Defensins; beta-Defensins; Blotting, Western; Enzyme-Linked Immunosorbent Assay; Eosinophil-Derived Neurotoxin; Eosinophilia; Female; Humans; Lactoferrin; Male; Muramidase; Nasal Lavage Fluid; Recurrence; Rhinitis, Allergic, Perennial; Ribonucleases; Sinusitis; Skin Tests

To distinguish sinusitis from uncomplicated "colds," we examined lactoferrin and eosinophilic cationic protein (ECP) in nasal secretions. Lactoferrin titers were >/=1:400 in 4% of persons with uncomplicated colds and controls but in 79% of persons with sinusitis or purulent sputa. ECP levels were >200 ng/ml in 61% of persons with colds and >3,000 ng/ml in 62% of persons with sinusitis. Nasal lactoferrin helps distinguish sinusitis from colds.

Topics: Bacterial Infections; Blood Proteins; Common Cold; Community-Acquired Infections; Eosinophil Granule Proteins; Humans; Lactoferrin; Mucus; Nasal Mucosa; Rhinovirus; Ribonucleases; Sinusitis

To study the local protective role of lysozyme(LZ) and lactoferrin(LF) in the uncinate process mucosa during chronic sinusitis.. Expression of LZ and LF was determined in 17 samples from normal subjects and 70 samples from chronic sinusitis patients with ABC immunohistochemical method. According to the presence or absence of nasal polyps, patients were divided into two groups.. Serous cells of submucosal glands displayed a strongly positive staining reaction to both LZ and LF in the normal uncinate process mucosa and mucosa from patients with chronic sinusitis. A positive though weak staining for LZ could also be found frequently within mucous cells of submucosal mixed glands and occasionally within goblet cells. In the mucosa from patients without nasal polyps, the staining reaction to LZ appeared to be intensified in goblet cells when compared with normal controls (P < 0.05). In patients with nasal polyps, the staining reaction to LZ appeared to be intensified in submucosal glands when compared with normal controls (P < 0.01) and patients without nasal polyps (P < 0.05). For LF, the staining reaction from patients with nasal polyps was stronger than that in normal controls (P < 0.01). The epithial cells stained negatively for LZ and LF.. It suggests that the observed increase in LZ and LF secreting activity of goblet cells and submucosal mixed glands may play a part role in the defense mechanism of uncinate process mucosa during the course of chronic sinusitis.

Topics: Adolescent; Adult; Child; Chronic Disease; Ethmoid Sinus; Female; Humans; Lactoferrin; Male; Middle Aged; Muramidase; Nasal Mucosa; Nasal Polyps; Sinusitis

Recurrent sinusitis (RS) is a very common clinical problem for which no underlying cause can generally be ascertained. We examined nasal mucosal responses in 14 patients with RS to determine if a relative deficiency in secretion of glandular antimicrobial factors might play a role. Twenty-four subjects with no history of sinusitis were studied concurrently as normal control (NC) subjects. RS was defined by two or more episodes of acute sinusitis per year for 2 or more years. After provocation with 25 mg of methacholine or 1 mg of histamine, nasal washings were analyzed for total proteins: the plasma protein albumin, IgG, and nonsecretory IgA (nsIgA), and the glandular proteins secretory IgA (sIgA), lactoferrin (LFN), and lysozyme (LZM). Although baseline secretions in patients with RS were relatively enriched with LFN and LZM as compared to that of secretions in NC subjects, patients with RS had a blunted cholinergic response with decreased secretion of albumin, IgG, nsIgA, sIgA, and LZM. Histamine responses were equivalent in both patients with RS and NC subjects. After 4 to 12 months of medical treatment, the abnormal cholinergic responses improved on repeat methacholine challenge in all eight subjects with RS rechallenged. Thus, patients with RS have a reversible reduction in nasal mucosal secretory responses to cholinergic stimulation. Since glandular secretions are rich in antimicrobial factors, such as LFN, LZM, and sIgA, it appears possible that the inability to secrete glandular proteins normally may predispose to recurrent infections.

Topics: Acute Disease; Adult; Female; Humans; Immunoglobulin A; Immunoglobulin A, Secretory; Immunoglobulin G; Lactoferrin; Male; Middle Aged; Muramidase; Nasal Mucosa; Nasal Provocation Tests; Recurrence; Serum Albumin; Sinusitis

Retained maxillary sinus secretions from 10 consecutive patients suffering from maxillary sinusitis were studied with regard to proteolytic activity and its possible sources. All secretions were proteolytically active. In 3 purulent secretions the proteolytic activity was of the same magnitude as that of a standard with an excess of pancreatic trypsin. The enzymes responsible for the proteolytical activity were found to be mainly of granulocyte origin, neutrophil elastase, unspecific collagenase and chymotrypsin-like cationic protein (CCP).

Topics: Adult; Aged; alpha 1-Antitrypsin; Blood Proteins; Female; Granulocytes; Humans; Immunoelectrophoresis; Lactoferrin; Male; Maxillary Sinus; Microbial Collagenase; Middle Aged; Pancreatic Elastase; Peptide Hydrolases; Sinusitis