Dynamic Interaction Between Mucosal Immunity and Microbiota Drives Nose and Pharynx Homeostasis of Common Carp (Cyprinus carpio) After SVCV Infection.

The crosstalk between the immune system and microbiota drives an amazingly complex mutualistic symbiosis. In mammals, the upper respiratory tract acts as a gateway for pathogen invasion, and the dynamic interaction between microbiota and mucosal immunity on its surface can effectively prevent disease development. However, the relationship between virus-mediated mucosal immune responses and microbes in lower vertebrates remains uncharacterized. In this study, we successfully constructed an infection model by intraperitoneally injecting common carp (Cyprinus carpio) with spring viremia of carp virus (SVCV). In addition to the detection of the SVCV in the nose and pharynx of common carp, we also identified obvious histopathological changes following viral infection. Moreover, numerous immune-related genes were significantly upregulated in the nose and pharynx at the peak of SVCV infection, after which the expression levels decreased to levels similar to those of the control group. Transcriptome sequencing results revealed that pathways associated with bacterial infection in the Toll-like receptor pathway and the Nod-like receptor pathway were activated in addition to the virus-related Rig-I-like receptor pathway after SVCV infection, suggesting that viral infection may be followed by opportunistic bacterial infection in these mucosal tissues. Using 16S rRNA gene sequencing, we further identified an upward trend in pathogenic bacteria on the mucosal surface of the nose and pharynx 4 days after SVCV infection, after which these tissues eventually reached new homeostasis. Taken together, our results suggest that the dynamic interaction between mucosal immunity and microbiota promotes the host to a new ecological state.

Infrahyoid myocutaneous flap: A systematic review.

INTRODUCTION: The infrahyoid myocutaneous flap (IHMCF) is an often-overlooked flap of the anterior neck used for reconstruction of oral cavity and laryngopharyngeal defects. The primary goal of this systematic review is to evaluate the postoperative outcomes and efficacy of this flap. METHODS: A comprehensive search of PubMed, Biological Abstracts, CINAHL Plus, and Web of Science was conducted. Two researchers independently scrutinized the studies to determine inclusions based on relevance, sample size, and English language publications. RESULTS: Twenty-eight studies containing 1027 IHMCF cases met the inclusion criteria. Primary outcomes included flap necrosis and postoperative functional outcomes. The rate of flap survival was 99%. Total skin necrosis and partial skin necrosis were minor complications that occurred in 2.5% and 5.8% of cases respectively. Poor speech and swallowing outcomes were reported in 6.4% and 6.5% of cases respectively. The included studies were predominantly retrospective. An average MINORS score of 9.6 suggests moderate bias among the studies. CONCLUSIONS: The IHMCF is both safe and effective for repairing medium sized mucosal lesions of the head and neck region in carefully selected patients. IHMCF use in oral cavity reconstruction is particularly appealing although functional outcomes remain difficult to statistically assess. Complications of IHMCFs are rare and often minor. To ensure the best outcome, pre-surgical planning needs to be conducted and all contraindications should be respected. Further large prospective multi-centered trials are needed for more accurate analysis.

Age-Associated Role of Lingual and Pharyngeal Glands in Local Immunity in Oral Cavity.

The macroscopic and histological methods were employed to examine the autopsy specimens of salivary lingual glands obtained from 299 patients of both sexes and various age ranging from newborn to longevity. The age-associated alterations of minor lingual and pharyngeal glands were revealed, and the topographical relations between the glands and lymphoid cells were described. The characteristic sparsity of the glands in infancy is caused by nutritional uniformity at this period, when diminished production of secretory IgA results in frequent inflammatory processes in oral and pharyngeal cavities. With age, the glandular orifices widen, and their number increases thereby augmenting local immunity in the oral cavity and in oral aspect of the pharynx. Starting from elderly and senile age, the involutive alterations were observed, which were accompanied by diminished production of secretory immunoglobulin A and related degradation of local and humoral immunity.

Microfluidic Immunoassays for Sensitive and Simultaneous Detection of IgG/IgM/Antigen of SARS-CoV-2 within 15 min.

The outbreak of SARS-CoV-2 is posing serious global public health problems. Facing the emergence of this pandemic, we established a portable microfluidic immunoassay system for easy-to-use, sensitive, rapid (<15 min), multiple, and on-site detection of IgG/IgM/Antigen of SARS-CoV-2 simultaneously. This integrated method was successfully applied for detecting SARS-CoV-2 IgM and IgG antibodies in clinical human serum as well as SARS-CoV-2 antigen in pharyngeal swabs from 26 patients with COVID-19 infection and 28 uninfected people. The assay demonstrated high sensitivity and specificity, which is promising for the diagnosis and monitoring as well as control of SARS-CoV-2 worldwide.

The epipharynx-kidney axis triggers glomerular vasculitis in immunoglobulin A nephropathy.

Macroscopic hematuria concomitant with acute pharyngitis is a characteristic feature of immunoglobulin A nephropathy (IgAN). Although the underlying mechanism of worsening hematuria has not been fully elucidated, activation of the innate immune system of nasopharynx-associated lymphoid tissue is thought to play an important role. The epipharynx is an immunologically activated site even under normal conditions, and enhanced activation of innate immunity is likely to occur in response to airborne infection. As latent but significant epipharyngitis presents in most IgAN patients, it is plausible that acute pharyngitis due to airway infection may contribute as a trigger of the epipharyngeal innate immune system, which is already upregulated in the chronically inflamed environment. The aim of this review was to discuss the mechanism of epipharynx-kidney axis involvement in glomerular vasculitis responsible for the worsening of hematuria in IgAN.

Controlled human infection for vaccination against Streptococcus pyogenes (CHIVAS): Establishing a group A Streptococcus pharyngitis human infection study.

Group A Streptococcus (GAS) is a highly-adapted and human-restricted pathogen responsible for a high global burden of disease across a diverse clinical spectrum. Vaccine development has been impeded by scientific, regulatory, and commercial obstacles. Human infection studies (HIS) are increasingly contributing to drug, diagnostics, and vaccine development, reducing uncertainty at early stages, especially for pathogens with animal models that incompletely reproduce key elements of human disease. We review the small number of historical GAS HIS and present the study protocol for a dose-ranging inpatient study in healthy adults. The primary objective of the study is to establish a new GAS pharyngitis HIS with an attack rate of at least 60% as a safe and reliable platform for vaccine evaluation and pathogenesis research. According to an adaptive dose-ranging study design, emm75 GAS doses manufactured in keeping with principles of Good Manufacturing Practice will be directly applied by swab to the pharynx of carefully screened healthy adult volunteers at low risk of severe complicated GAS disease. Participants will remain as closely monitored inpatients for up to six days, observed for development of the primary outcome of acute symptomatic pharyngitis, as defined by clinical and microbiological criteria. All participants will be treated with antibiotics and followed as outpatients for six months. An intensive sampling schedule will facilitate extensive studies of host and organism dynamics during experimental pharyngitis. Ethics approval has been obtained and the study has been registered at ClinicalTrials.gov (NCT03361163).

The Axial Organ and the Pharynx Are Sites of Hematopoiesis in the Sea Urchin.

Background: The location of coelomocyte proliferation in adult sea urchins is unknown and speculations since the early 1800s have been based on microanatomy and tracer uptake studies. In adult sea urchins (Strongylocentrotus purpuratus) with down-regulated immune systems, coelomocyte numbers increase in response to immune challenge, and whether some or all of these cells are newly proliferated is not known. The gene regulatory network that encodes transcription factors that control hematopoiesis in embryonic and larval sea urchins has not been investigated in adults. Hence, to identify the hematopoietic tissue in adult sea urchins, cell proliferation, expression of phagocyte specific genes, and expression of genes encoding transcription factors that function in the conserved regulatory network that controls hematopoiesis in embryonic and larval sea urchins were investigated for several tissues. Results: Cell proliferation was induced in adult sea urchins either by immune challenge through injection of heat-killed Vibrio diazotrophicus or by cell depletion through aspiration of coelomic fluid. In response to either of these stimuli, newly proliferated coelomocytes constitute only about 10% of the cells in the coelomic fluid. In tissues, newly proliferated cells and cells that express SpTransformer proteins (formerly Sp185/333) that are markers for phagocytes are present in the axial organ, gonad, pharynx, esophagus, and gut with no differences among tissues. The expression level of genes encoding transcription factors that regulate hematopoiesis show that both the axial organ and the pharynx have elevated expression compared to coelomocytes, esophagus, gut, and gonad. Similarly, an RNAseq dataset shows similar results for the axial organ and pharynx, but also suggests that the axial organ may be a site for removal and recycling of cells in the coelomic cavity. Conclusions: Results presented here are consistent with previous speculations that the axial organ may be a site of coelomocyte proliferation and that it may also be a center for cellular removal and recycling. A second site, the pharynx, may also have hematopoietic activity, a tissue that has been assumed to function only as part of the intestinal tract.

Anti-inflammatory activity of resveratrol prevents inflammation by inhibiting NF­κB in animal models of acute pharyngitis.

Recent studies have demonstrated that resveratrol can reduce blood sugar, improve insulin resistance, regulate abnormalities in lipid metabolism, and lower the secretion and expression of inflammatory factors. The present study investigated the anti­inflammatory effects of resveratrol in animal models of acute pharyngitis, and its possible mechanisms. Commercial ELISA kits were used to measure tumor necrosis factor­α, interleukin (IL)­6, macrophage inflammatory protein­2, cyclooxygenase­2 levels and caspase­3/9 activity. Toll­like receptor (TLR)­4, myeloid differentiation primary response protein MyD88, phosphorylated (p)­nuclear factor (NF)­κB and p­IκB were analyzed using western blotting. In a rabbit model of acute pharyngitis, it was demonstrated that resveratrol inhibited tumor necrosis factor­α and interleukin­6 serum levels, macrophage inflammatory protein­2 and cyclooxygenase­2 activity levels, reactive oxygen species production and caspase­3/9 activity. Resveratrol suppressed NACHT, LRR and PYD domains­containing protein 3 and caspase­1 protein expression, and reduced IL­1ß and IL­18 protein expression in animal models of acute pharyngitis. Additionally, resveratrol suppressed TLR4 and myeloid differentiation primary response protein 88 protein expression, and reduced p­NF­κB and increased p­IκB protein expression in animal models of acute pharyngitis. In conclusion, these findings indicated that the anti­inflammatory activity of resveratrol prevents acute pharyngitis­induced inflammation by inhibiting NF­κB in animal models. Therefore, these data suggested an important clinical application of resveratrol in preventing acute pharyngitis.

In vitro assessment of the ability of probiotics, blueberry and food carbohydrates to prevent S. pyogenes adhesion on pharyngeal epithelium and modulate immune responses.

Group A streptococci (GAS) cause 20-30% of pediatric pharyngitis episodes, which are a major cause of ambulatory care visits. Therefore, a strategy to prevent GAS dissemination in children could significantly benefit public healthcare. Contextually, we assessed the possibility of employing alternative food-grade strategies to be used with the oral probiotic L. helveticus MIMLh5 for the prevention of pharyngeal infections. First, we demonstrated through an antagonism-by-exclusion assay that guaran may potentially prevent S. pyogenes adhesion on pharyngeal cells. Subsequently, we showed that an anthocyanin-rich fraction extracted from wild blueberry (BbE) exerts anti-inflammatory effects on the human macrophage cell line U937. Finally, we showed that BbE reduces interferon-ß expression in MIMLh5-stimulated murine dendritic cells, resulting in a reduction in the pro-inflammatory cytokines IL-12 and TNF-α. In conclusion, this proof-of-concept study indicates that different food-grade strategies may be concomitantly adopted to potentially prevent GAS colonization and modulate local immune defences.

Incremental Contributions of FbaA and Other Impetigo-Associated Surface Proteins to Fitness and Virulence of a Classical Group A Streptococcal Skin Strain.

Group A streptococci (GAS) are highly prevalent human pathogens whose primary ecological niche is the superficial epithelial layers of the throat and/or skin. Many GAS strains with a strong tendency to cause pharyngitis are distinct from strains that tend to cause impetigo; thus, genetic differences between them may confer host tissue-specific virulence. In this study, the FbaA surface protein gene was found to be present in most skin specialist strains but largely absent from a genetically related subset of pharyngitis isolates. In an ΔfbaA mutant constructed in the impetigo strain Alab49, loss of FbaA resulted in a slight but significant decrease in GAS fitness in a humanized mouse model of impetigo; the ΔfbaA mutant also exhibited decreased survival in whole human blood due to phagocytosis. In assays with highly sensitive outcome measures, Alab49ΔfbaA was compared to other isogenic mutants lacking virulence genes known to be disproportionately associated with classical skin strains. FbaA and PAM (i.e., the M53 protein) had additive effects in promoting GAS survival in whole blood. The pilus adhesin tip protein Cpa promoted Alab49 survival in whole blood and appears to fully account for the antiphagocytic effect attributable to pili. The finding that numerous skin strain-associated virulence factors make slight but significant contributions to virulence underscores the incremental contributions to fitness of individual surface protein genes and the multifactorial nature of GAS-host interactions.

The Ciona intestinalis immune-related galectin genes (CiLgals-a and CiLgals-b) are expressed by the gastric epithelium.

The transcription of two Ciona intestinalis galectin genes (CiLgals-a and CiLgals-b) is uparegulated by LPS in the pharynxis (hemocytes, vessel epithelium, endostilar zones) which is retained the main organ of the immunity. In this ascidian, for the first time we show, by immunohistochemistry and in situ hybridization methods, that these two immune-related genes are expressed in the gastric epithelium of naïve ascidians, whereas the galectins appear to be only contained in the intestine columnar epithelium. In addition, according to previous results on the pharynx, the genes are also expressed and galectins produced by hemocytes scattered in the connective tissue surrounding the gut. The genes expression and galectin localization in several tissues, including the previous findings on the transcription upregulation, the constitutive expression of these genes by endostylar zones and by the gastric epithelium suggest a potential multifunctional role of these galectins. In this respect, it is of interest to define where the CiLgals are normally found as related to the tissue functions. Such an approach should be a starting point for further investigations.

Analysis of clinical and immunophenotypic features along with treatment outcomes of diffuse large B cell lymphoma patients, based on the involvement of nodal or extranodal primary sites.

AIMS: The aim of the present study was to elucidate the clinical features, immunophenotype and treatment outcomes of diffuse large B cell lymphoma (DLBCL) patients based on the involvement of the primary site i.e. lymph node or specific extranodal sites. METHODS: We analyzed the clinical characteristics, immunophenotype and treatment outcomes of 207 DLBCL patients diagnosed in China between 2007 and 2014 based on the primary site of location. RESULTS: Based on the involvement of primary site of occurrence, DLBCL cases, were classified into different groups like, lymph node (60 cases, 28.98%), gastrointestinal tract (GI) (53 cases, 25.60%), Waldeyer's ring (WR) (31 cases, 14.97%), gland (25 cases, 12.08%), and other extranodal sites (38 cases, 18.36%). Patients with WR involvement were more frequently associated with early stage disease, favorable performance status, absence of bulky disease, normal LDH and ESR levels, and low- or low/intermediate-risk IPI than the other groups. The proportion of DLBCL patients with germinal center B cell (GCB) phenotype was 56.0% for WR, 46.5% for GI, 34.5% for lymph node, 27.8% for other extranodal sites, and 18.2% for gland (P=0.035). The 5-year overall survival (OS) of the entire patient population was 71.1%, and WR group showed a better outcome than nodal group (84.9% vs. 55.9%, P=0.015). In multivariate analysis, bulky disease, bone marrow infiltration, non-GCB phenotype, intermediate/high- or high-risk IPI and SD/PD/Death after first therapy were identified as independent factors for poor OS, while regular application of rituximab and remission after first therapy were identified as favorable prognostic factors for PFS. CONCLUSIONS: In this study, WR involvement was associated with more favorable clinical & pathological features along with better outcome than nodal lymphoma. The OS and PFS were largely dependent on other prognostic variables such as IPI or immunophenotype instead of the sites of involvement.

RANKL regulates differentiation of microfold cells in mouse nasopharynx-associated lymphoid tissue (NALT).

Murine nasopharynx-associated lymphoid tissue (NALT), located at the base of the nasal cavity, serves as a major site for the induction of mucosal immune responses against airway antigens. The follicle-associated epithelium (FAE) covering the luminal surface of NALT is characterized by the presence of microfold cells (M cells), which take up and transport luminal antigens to lymphocytes. Glycoprotein 2 (GP2) has recently been identified as a reliable marker for M cells in Peyer's patches of the intestine. However, the expression of GP2 and other functional molecules in the M cells of NALT has not yet been examined. We have immunohistochemically detected GP2-expressing cells in the FAE of NALT and the simultaneous expression of other intestinal M-cell markers, namely Tnfaip2, CCL9, and Spi-B. These cells have been further identified as M cells because of their higher uptake capacity of luminal microbeads. Electron microscopic observations have shown that GP2-expressing cells on the FAE display morphological features typical of M cells: they possess short microvilli and microfolds on the luminal surface and are closely associated with intraepithelial lymphocytes. We have also found that the receptor activator of nuclear factor kappa-B ligand (RANKL) is expressed by stromal cells underneath the FAE, which provides its receptor RANK. The administration of RANKL markedly increases the number of GP2(+)Tnfaip2(+) cells on the NALT FAE and that of intestinal M cells. These results suggest that GP2(+)Tnfaip2(+) cells in NALT are equivalent to intestinal M cells, and that RANKL-RANK signaling induces their differentiation.

EV71 infection correlates with viral IgG preexisting at pharyngo-laryngeal mucosa in children.

Enterovirus 71 (EV71) infection causes severe central nervous system damage, particularly for children under the age of 5 years old, which remains a major public health burden worldwide. Clinical data released that children may be repeatedly infected by different members in enterovirus and get even worsen. Mucosa, especially epithelium of alimentary canal, was considered the primary site of EV71 infection. It has been elusive whether the preexsiting viral antibody in mucosa plays a role in EV71 infection. To answer this question, we respectively measured viral antibody response and EV71 RNA copy number of one hundred throat swab specimens from clinically confirmed EV71-infected children. The results released that low-level of mucosal IgG antibody against EV71 broadly existed in young population. More importantly, it further elucidated that the children with mucosal preexsiting EV71 IgG were prone to be infected, which suggested a former viral IgG mediated enhancement of viral infection in vivo.

Oral, nasal and pharyngeal exposure to lipopolysaccharide causes a fetal inflammatory response in sheep.

BACKGROUND: A fetal inflammatory response (FIR) in sheep can be induced by intraamniotic or selective exposure of the fetal lung or gut to lipopolysaccharide (LPS). The oral, nasal, and pharyngeal cavities (ONP) contain lymphoid tissue and epithelium that are in contact with the amniotic fluid. The ability of the ONP epithelium and lymphoid tissue to initiate a FIR is unknown. OBJECTIVE: To determine if FIR occurs after selective ONP exposure to LPS in fetal sheep. METHODS: Using fetal recovery surgery, we isolated ONP from the fetal lung, GI tract, and amniotic fluid by tracheal and esophageal ligation and with an occlusive glove fitted over the snout. LPS (5 mg) or saline was infused with 24 h Alzet pumps secured in the oral cavity (n = 7-8/group). Animals were delivered 1 or 6 days after initiation of the LPS or saline infusions. RESULTS: The ONP exposure to LPS had time-dependent systemic inflammatory effects with changes in WBC in cord blood, an increase in posterior mediastinal lymph node weight at 6 days, and pro-inflammatory mRNA responses in the fetal plasma, lung, and liver. Compared to controls, the expression of surfactant protein A mRNA increased 1 and 6 days after ONP exposure to LPS. CONCLUSION: ONP exposure to LPS alone can induce a mild FIR with time-dependent inflammatory responses in remote fetal tissues not directly exposed to LPS.

Ciona intestinalis galectin (CiLgals-a and CiLgals-b) genes are differentially expressed in endostyle zones and challenged by LPS.

Immunohistochemical and in situ hybridization assays were performed to answer the question whether the endostyle, that is the initial gastro-intestinal trait of Ciona intestinalis pharynx, is involved in galectin (CiLgals-a and CiLgals-b) production during the pharynx inflammatory response to LPS inoculation. Specific anti-CiLgal-a and anti-CiLgals-b antibodies, and oligonucleotide probes, that mark inflammatory hemocytes inside the pharynx vessels and vessel epithelium as shown by a previous paper, were assayed on endostyle histological sections. For the first time, we show that galectins are produced by endostyle zones, and both CiLgals-a and -b genes are upregulated by LPS. CiLgals-a and CiLgals-b are constitutively expressed in the endostyle zone 2 and 3, respectively, both genes are upregulated by LPS in the zone 2, and CiLgals-b in the zone 3 and 4. The antibody-reacting material contained in intracellular and extracellular large vesicles suggest an unexpected vesicle-dependent transporting mechanism of galectins not provided with signal peptide. Differential expression and gene upregulation in not-treated and LPS-treated specimens, support the role of endostyle galectins both in filter feeding and defense responses.

AAOA allergy primer: history and physical examination.

BACKGROUND: Allergic disease is very common in the general population and makes a significant impact on the quality of life of patients. Immunoglobulin E (IgE)-mediated allergic disease manifests throughout the body, but many signs and symptoms of inhalant allergy are centered in the head and neck region. METHODS: A thorough yet focused history of allergic symptoms and potential physical examination findings of inhalant allergy are described. RESULTS: History should include types and timing of symptoms, environmental and occupational exposures, family history, associated diseases, and prior treatment, if any. Physical examination should include the skin and structures of the head and neck region. Nasal endoscopy can be helpful in visualization of nasal polyps. CONCLUSION: Many times, history alone can serve to make the diagnosis, but physical examination also demonstrates specific findings that confirm the practitioner's presumptive diagnosis of allergic disease. However, should medical treatment fail or the diagnosis be in doubt, further diagnostic investigation with allergy testing should be pursued.

The mucosal immune system in the oral cavity-an orchestra of T cell diversity.

The mucosal immune system defends against a vast array of pathogens, yet it exhibits limited responses to commensal microorganisms under healthy conditions. The oral-pharyngeal cavity, the gateway for both the gastrointestinal and respiratory tracts, is composed of complex anatomical structures and is constantly challenged by antigens from air and food. The mucosal immune system of the oral-pharyngeal cavity must prevent pathogen entry while maintaining immune homeostasis, which is achieved via a range of mechanisms that are similar or different to those utilized by the gastrointestinal immune system. In this review, we summarize the features of the mucosal immune system, focusing on T cell subsets and their functions. We also discuss our current understanding of the oral-pharyngeal mucosal immune system.

Detection of chlamydophila pneumoniae antigens in children in the lower silesia region in 2011.

BACKGROUND: The majority of Chlamydophila pneumoniae infections are asymptomatic. Symptomatic infections due to C. pneumoniae are associated with upper and lower respiratory tract infections. OBJECTIVES: Analysis of the frequency of C. pneumoniae antigens detection in children in the Lower Silesia Region in 2011. MATERIAL AND METHODS: 303 throat swabs obtained from 130 girls and 173 boys, aged 20 months to 18 years, were tested. The patients who were entered into the study were treated due to various respiratory disorders. Examinations were performed by IFA technique, using the Chlamydia Cell PN testing kits (Cellabs Pty Ltd., Sydney, Australia). RESULTS: The presence of C. pneumoniae antigen was detected in 95/303 (31.4%) of all examined children (in 48/130 (36.9%) of girls and in 47/173 (27.2%) of boys). In the group of patients with dry cough, as the most common presenting clinical symptom of respiratory infection, the positive IFA test results for C. pneumoniae occurred in 32 out of 94 (34.0%) of girls and in 34 out of 117 (29.1%) of boys. In the case of the examined children with other prevailing clinical symptom, which included a cough with discharge and a runny nose, the positive tests for C. pneumoniae were shown in 13 out of 27 (48.1%) of girls and in 9 out of 42 (21.4%) of boys. In the group of children without symptoms of infection, who had direct contact with C. pneumoniae infected person, there were 3 out of 9 (33.3%) of girls and 4 out of 14 (28.6%) of boys positively tested cases. CONCLUSIONS: Results of IFA studies for C. pneumoniae in throat swabs in children varies according to clinical symptoms and seasonality. There were no clear gender and age-related differences. C. pneumoniae test should be considered only after the presence of microorganisms typical of respiratory infections has been excluded.