Article

Moving Towards a New Era in the Research of Tonsils and Mucosal Barriers

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Abstract

The palatine and nasopharyngeal tonsils (adenoids) are lymphoepithelial tissues located in strategic anatomical areas of the oral pharynx and nasopharynx. These immunocompetent tissues represent the first line of defense against ingested or inhaled foreign proteins such as bacteria, viruses, or food antigens. Accompanying the advances being made in the field of medicine today, the role of the tonsils in immunocompetence is becoming extremely important. Upper respiratory tract infections such as acute otitis media, acute rhinosinusitis and acute pharyngo-tonsillitis are diseases that occur with extremely high frequency, and the antimicrobial agents used to treat these diseases account for a large proportion of health care costs. The increasingly refractory nature of upper respiratory tract infections caused by drug-resistant bacteria has become a major worldwide concern. The elucidation of the immune functions of the tonsils and mucosal membranes of the upper respiratory tract is considered to have important significance. The tonsils are also considered to play an important role as one of the causes of sleep apnea syndrome, and have been reported to be intimately involved in the manifestation of IgA nephropathy and palmoplantar pustulosis, a kind of skin disorder. Interest has continued to grow in this symposium with each session ever since it was first held in Kyoto, Japan in 1987. Since then, the symposium has been held every 3-4 years; in Pavia in 1991, in Sapporo in 1995, in Ghent in 1999, in Wakayama in 2003, and in Siena in 2006. Since the 5th symposium in Wakayama, the topics were extended to mucosal barriers of upper airways including the mucosal immune system, innate immunity, and mucosal vaccine. Recent fine technologies and information on molecular biological approaches for upper airways will continue to advance our understanding of epidemiology, etiology, pathogenesis, diagnosis and management of tonsil-related disorders and various upper respiratory tract infections such as otitis media and rhinosinusitis. Moreover, in the era of drug-resistant microbes, we should exert more effort to develop powerful and effective mucosal vaccines against pathogens in upper airways.

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... Дані досліджень з теоретичних аспектів функціонування піднебінних мигдаликів в умовах їхньої гіпертрофії та хронічного запалення можуть сприяти створенню нових технологій лікування цих процесів у дитячому віці. Відомо, що структурні і функціональні зміни в піднебінних мигдалинах дітей при гіпертрофії та хронічному запаленні відрізняються за клітинною щільністю тканини, кількістю сполучнотканинних елементів [4,5], активністю функціональних груп імунокомпетентних клітин, цитокінопродукціі, включаючи інтерферони [6][7][8]. Аналізуючи дані наукової літератури, неважко дійти висновку, що функціональний стан клітин піднебінних мигдаликів при зазначених захворюваннях є відображенням тих структурних змін, які відбуваються в самій їхній тканині. Це опосередковано підтверджується в роботах останніх років, в яких зроблені спроби визна- ...
... Дані досліджень з теоретичних аспектів функціонування піднебінних мигдаликів в умовах їхньої гіпертрофії та хронічного запалення можуть сприяти створенню нових технологій лікування цих процесів у дитячому віці. Відомо, що структурні і функціональні зміни в піднебінних мигдалинах дітей при гіпертрофії та хронічному запаленні відрізняються за клітинною щільністю тканини, кількістю сполучнотканинних елементів [4,5], активністю функціональних груп імунокомпетентних клітин, цитокінопродукціі, включаючи інтерферони [6][7][8]. Аналізуючи дані наукової літератури, неважко дійти висновку, що функціональний стан клітин піднебінних мигдаликів при зазначених захворюваннях є відображенням тих структурних змін, які відбуваються в самій їхній тканині. Це опосередковано підтверджується в роботах останніх років, в яких зроблені спроби визна-чити ступінь ураження тканини піднебінних мигдаликів при гіпертрофії та хронічному запаленні та провести корелятивні зв'язки з відображенням основних відхилень в системному імунітеті для обґрунтування вибору методу лікування -консервативного або оперативного [9]. ...
... In most cases, chronic inflammatory diseases of the upper respiratory tract of both microbial and viral origin occur due to the immune deficiency of systemic and local nature [2,12,14,15]. Among a large number of immunomodulation agents, a special role belongs to plant preparations, which are characterized by relative harmlessness and high efficiency, both in case of systemic and local use [9,12]. ...
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Introduction: Chronic inflammatory diseases of the upper respiratory tract of both microbial and viral origin occur due to the immune deficiency of systemic and local nature. Among a large number of immunomodulation agents, a special role belongs to plant preparations, which are characterized by relative harmlessness and high efficiency, both in case of systemic and local use and they are often combined with antimicrobial agents, therefore the aim of the study was to investigate their effect on the reactions and condition of lymphoid cells of tonsilswith separate and combined use in vitro. Methods: Cell suspensions were prepared mechanically and adjusted to a concentration of 2 million/ml in medium 199 with additives. Then, Lizak preparation was added to the cells and starch solution was used in control. Imupret and Esberitox preparations were added to the culture with or without Lizak preparation. After cultivation, the levels of proinflammatory cytokine – Interleukin-1β, pro-allergic factor – Interleukin-4, Th-1 – derivative of cytokine – interferon-γ were studied in the supernatant using the «Tsitokin LLC» reagent kits (RF), as well as tissue regeneration factor – transforming growth factor –TGF-1β (Austria). The preparation was prepared from the cell pellet, in which the relative nonviable cell count was determined in a sample with trypan blue using a light microscope (Olympus CX21FS1).The statistical analysis was performed using Student's t-distribution. Results: Plant-derived preparations did not have any effect on cell viability in culture, did not reduce the level of the pro-inflammatory cytokine interleukin-1β and did not stimulate the production of the regeneration factor TGF-1β. At the same time, the combined use of plant-derived preparations and the antibacterial preparation Lizak in cell culture of tonsils was accompanied by an increase in cell viability compared to the use of Lizak preparation separately, the stimulation of the regulatory antiviral factor interferon-γ and an increase in the production of tissue regeneration factor. Conclusion: The immunomodulating properties of the antibacterial preparation Lizak and plant-derived immunomodulator Esberitox significantly differ in the points of application. Both plant-derived preparations stimulated the production of γ-interferon by tonsil cells in the presence of Lizak preparation, the transforming growth factor TGF-1β, the viability of tonsil cells was the highest with the combination of Lizak and Esberitox.
... В настоящее время считается доказанным, что инфекционно-воспалительные заболевания верхних дыхательных путей как микробного, так и вирусного генеза возникают и поддерживаются за счет иммунной недостаточности как системного, так и, особенно, локального характера [4,7,11,12]. В связи этим важную роль играет правильное и своевременное применение методов и способов иммунокоррекции, основанных на современной и адекватной диагностике состояния факторов локального иммунитета в ротоглоточном секрете [6,9,10]. ...
Article
Introduction: Among a large number of immunomodulation agents, a special role belongs to herbal medicines. They differ in relative harmlessness and high efficiency, both with system and local use. Clinical and immunological efficacy of Esberitox phytopreparation (Schaper&Brummer, Germany) in a number of diseases of viral and microbial origin has been shown. This report presents data on the effect of Esberitox on the factors of local immunity of the oral cavity in patients with chronic pharyngitis, tonsillitis and periodontitis with systemic use of Esberitox in monotherapy. Methods: The oropharyngeal secret of 10 patients with chronic tonsillitis (CT), 11 patients with chronic pharyngitis (CP), (all patients after tonsillectomy), 10 patients with chronic periodontitis was investigated. Patients were given monotherapy with «Esberitox» for 10 days (CP), according to the manufacturer's instructions for use of the drug. In addition, 9 practically healthy persons (K) of the same age (18-50 years) were examined. The ELISA method was used to determine the class A immunoglobulins (secretory and monomeric), IgG, α-interferon, interleukin-1β, levels of immune complexes. Statistics: Mann-Whitney U-test. Results: Studies have shown that in patients with chronic inflammatory diseases of the oral cavity there is a decrease in the protective factors of local immunity – the level of secretory IgA, antiviral factor – α-interferon and an increase in the level of pro-inflammatory cytokine – interleukin-1β, monomeric form of IgA and the number of immune complexes. The use of «Esberitox» as a monotherapy was accompanied by an increase in the concentration of class A secretory immunoglobulin, the «normalization» of the α-interferon content (increase in concentration) and interleukin-1β (decrease in content). Conclusion: The resulting positive effects from the use of esberitox allow us to recommend the drug as part of the complex therapy of chronic diseases of the oropharynx, and in the form of monotherapy.
... Isolates of H. influenzae can subdivide into two important forms; encapsulated and non-encapsulated that serotype b causes meningitis, mainly in children below four years of age (Resman et al., 2011). Nontypeable H. influenzae (NTHi) usually is a commensal with the capability of producing infections of the upper and lower respiratory tracts, such as sinusitis, bronchitis acute otitis media, as well as, more infrequently, severe invasive infections such as pneumonia, bacteremia, and meningitis (Clementi and Murphy, 2011;Yamanaka, 2011). ...
... This ring belongs to the mucosa-associated lymphoid tissue (MALT) (Pabst & Brandzaeg, 2013). The nasopharyngeal tonsil, which is situated at the roof of the nasopharynx, represents the first line of defense at this strategic location against invading antigens that pass through the aero-digestive tract (Yamanaka, 2011). The tonsil has been described in many species (Chakar, 2016;Liebler-Terino & Pabst, 2006). ...
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The light microscopic appearance and ultrastructure of the nasopharyngeal tonsil (tonsilla pharyngea), collected from 12 adult buffaloes of local mixed breed, were explored for the distribution of different types of epithelia, lymphoid tissue and high endothelial venules. The tonsillar mucosa was lined by pseudostratified columnar ciliated epithelium having goblet cells. The respiratory epithelium associated with the underlying lymphoid tissue formed the lymphoepithelium. The epithelium was further modified into follicle‐associated epithelium (FAE) characterized by reduced epithelial height, presence of a few dome‐shaped cuboidal cells equivalent of the M‐cells and absence of goblet and ciliated cells. The lymphoid tissue was distributed in the form of isolated lymphoid cells, diffuse lymphoid tissue and lymphoid follicles, mainly distributed within the propria‐submucosa along with the sero‐mucous glandular tissue. The goblet cells of the respiratory epithelium and the acinar cells contained different mucopolysaccharides. Scanning electron microscopy of the surface mucosa demonstrated a dense mat of cilia, island‐like arrangement of microvillus cells, M‐cells and a few brush‐like cells. The transmission electron microscopy revealed the different cell organelles of the respiratory epithelium and the FAE. Lymphocyte migration via the high endothelial venules in the propria‐submucosa was also observed.
... Isolates of H. influenzae can subdivide into two important forms; encapsulated and non-encapsulated that serotype b causes meningitis, mainly in children below four years of age (Resman et al., 2011). Nontypeable H. influenzae (NTHi) usually is a commensal with the capability of producing infections of the upper and lower respiratory tracts, such as sinusitis, bronchitis acute otitis media, as well as, more infrequently, severe invasive infections such as pneumonia, bacteremia, and meningitis (Clementi and Murphy, 2011;Yamanaka, 2011). ...
... However, these bacteria can sometimes find their way to overcome the defense barriers of such locations and establish chronic infections that poses a challenge to both medical practice and healthcare system [2]. Infections of the ENT such as tonsillitis are diseases that occur with high frequency [3]. During the past decades, efforts have been made to manage the infectious diseases of tonsils [4]. ...
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Diseases of the tonsils are becoming more resistant to antibiotics due to the persistence of bacteria through the formation of biofilms. Therefore, understanding the microbiology and pathophysiology of such diseases represent an important step in the management of biofilm-related infections. We have isolated the microorganisms, evaluated their antimicrobial susceptibility, and detected the presence of bacterial biofilms in tonsillar specimens in correlation with the clinical manifestations of tonsillar diseases. Therefore, a total of 140 palatine tonsils were collected from 70 patients undergoing tonsillectomy at University Malaya Medical Centre. The most recovered isolate was Staphylococcus aureus (39.65%) followed by Haemophilus influenzae (18.53%). There was high susceptibility against all selected antibiotics except for cotrimoxazole. Bacterial biofilms were detected in 60% of patients and a significant percentage of patients demonstrated infection manifestation rather than obstruction. In addition, an association between clinical symptoms like snore, apnea, nasal obstruction, and tonsillar hypertrophy was found to be related to the microbiology of tonsils particularly to the presence of biofilms. In conclusion, evidence of biofilms in tonsils in correlation with the demonstrated clinical symptoms explains the recalcitrant nature of tonsillar diseases and highlights the importance of biofilm's early detection and prevention towards better therapeutic management of biofilm-related infections.
... Apesar dos avanços ocorridos nos diversos campos da Medicina, as infecções do trato respiratório superior como otite média aguda, rinossinusite aguda, faringotonsilites e as hipertrofias tonsilares são doenças que ocorrem com alta frequência e seu tratamento é responsável por grande parcela dos custos com a saúde 5 . ...
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Hypertrophy and recurrent tonsillitis are common indications of tonsillectomy. However, the pathological reports are similar, regardless of clinical aspects. Search for histopathological changes that differentiate palatine tonsils operated because of hypertrophy vis-à-vis those operated because of recurrent tonsillitis. A prospective cross-sectional descriptive study involving 46 children divided into group I - 22 with hypertrophy; and group II - 24 with recurrent tonsillitis, in the period between 2010 and 2012, in a public hospital. We evaluated clinical and histopathological features (lymph follicles, germinal centers, fibrosis, necrosis, reticulation, infiltration by plasma cells and neutrophils). The patients' ages ranged between 2 and 11 years (5.17 ± 2.28). In group I, half of the patients had had the latest infection at seven months or more, and all with obstruction degree greater than 3 (> 50%). In group II, all had had the latest infection at less than seven months, and most with obstruction degree below 4 (< 75%). There was a statistically significant difference in the degree of obstruction (p = 0.0021) and number of germinal centers (p = 0.002) was higher in group I. This study suggests that the number of germinal centers is the only histopathological criterion that can be used to differentiate the two groups.
... The selected samples were free of any inflammation or antibiotic therapy for at least one month, rendering them as close to normal as possible. If the samples were otherwise collected from autopsy of persons dying of other reasons, it would have been inappropriate to consider them normal in light of the fact that tonsillitis is a very common condition 11,12 , and an accurate history of the current status of tonsils is usually not available in these cases. ...
Article
To compare the thickness and organization of surface epithelium, and high endothelial venules in subepithelial compartments of human nasopharyngeal and palatine tonsils, with reference to functional differences. Comparative cross-sectional. The Anatomy Department of CPSP Regional Centre, Islamabad, from January to December 2005. Thirty samples each of human nasopharyngeal, right palatine and left palatine tonsils were collected by convenience sampling technique. Haematoxylin and eosin stained paraffin sections were examined for surface epithelium. Thickness of stratified squamous and pseudostratified ciliated columnar epithelium was measured, while organization was observed in case of stratified squamous. The high endothelial venules in subepithelial lymphoid compartments were counted. The surface epithelium of nasopharyngeal tonsils (stratified squamous and pseudostratified columnar inclusive) was 63.21+1.93 microm, and that of palatine (stratified squamous) was 143.99+5.94 microm thick (p<0.001). The mean count of high endothelial venules in subepithelial compartments of nasopharyngeal was 1.15+/-0.06 and that of palatine tonsil was 0.93+0.08 (p=0.042). Organization of stratified squamous epithelium was poor in 26 out of 30 nasopharyngeal, and well in all palatine tonsils (p<0.001). The surface epithelium of nasopharyngeal tonsil being thinner and poorly organized than that of palatine tonsil might act as a less effective barrier between the antigenic stimulus and subepithelial lymphoid compartments. This may contribute towards higher level of immune response by these compartments of the former, which is endorsed by higher number of high endothelial venules as compared to the latter.
Article
Objective: The aim: The aim of the study is to compare the class G antibody content in serum and tissue lysate from tonsils of children with hypertrophy and chronic tonsillitis to: streptolysin-O of Str. haemolyticus, protein-A of S. aureus, proteoglycans of Klebsiela spp., as well as to compare the content of interleukins 1β, 10, TNF-α, γ-IFN and lactoferrin in serum and tissue lysate from tonsils of children with hypertrophy and chronic tonsillitis. Patients and methods: Materials and methods: We studied tonsils of 33 children aged 4-18 years with hypertrophy of palatine tonsils (HPT) and with chronic tonsillitis (CT). The content of interleukins 1β, 10, TNF-α, γ-IFN and lactoferrin in tonsil lysate and serum was determined by immunofluorescence assay. Antistreptolysin O was studied by neutralization test of micromethod; class G antibodies to protein A of S. aureus and proteoglycans of Klebsiela spp. were studied by treponema pallidum hemagglutination assay. All the results were statistically processed using U-test (Mann-Whitney-Wilcoxon test) and Fisher's z-transformation. Results: Results: The serum and tissue lysate from tonsils of patients with HPT showed significantly high level of antibodies to streptolysin O in comparison with similar studies of substrates from patients with CT. Anti-inflammatory cytokine IL-10 was detected only in the serum of patients with CT. The TNF-α concentration in the lysates of tonsils in the group of patients with HPT was 2 times higher than in the group of patients with CT. The γ-IFN concentration was significantly lower both in the serum and in the lysates of tonsils of patients with CT. The content of lactoferrin in the lysates of patients with CT was 3 times higher (P<0.05) than in the lysates of patients with HPT. Conclusion: Conclusions: The results indicate a significant difference in the state of antibodies to microbial antigens and cytokines production in case of HPT and CT. In tonsils with HPT, there predominate reactions of antibody production to bacterial antigens and antiviral reactions like a high-level cytokines TNF-α and γ-IFN in tissue lysate of palatine tonsils.
Article
Background: Chronic tonsillitis (CT) is a common inflammatory illness in children, and serum antistreptolysin O titer (ASOT) is a common investigation performed for these cases and considered a perfect sign for tonsillectomy. Objective: To evaluate the expression of tonsillar T-and B-lymphocytes markers in relation to seropositive or seronegative ASOT in cases of CT. Materials and methods: Thirty children (15 males and 15 females) aged 6-10 years were divided equally into two groups: Group A seropositive ASOT (≥400 IU) and Group B seronegative ASOT (<400 IU). Both performed bilateral tonsillectomy. Specimens from the removed tonsils were taken and prepared for light microscopic examination and immunohistochemical evaluation of CD20 and CD3 expression. Results: Seropositive ASOT group showed significant histopathologic changes in the form of hyperplasia of the stratified squamous nonkeratinized epithelium, Urgas's abscess, and severe lymphocytic infiltration. Immunohistochemical results of seropositive ASOT group showed marked expression of CD3 and CD20, while seronegative ASOT group showed mild expression of CD3 and CD20. Conclusion: Seropositive ASOT CT, in addition to histopathological changes, is associated with significant increase in both B-lymphocytes (CD20 expression) and T-lymphocytes (CD3 expression) markers.
Chapter
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Two hundred and seventy-two strains of Streptococcus pyogenes isolated from patients with invasive and noninvasive infections in Japan were evaluated for the prevalence of fibronectin-binding protein genes (prtF1 and prtF2). The possible associations of the genes with streptococcal pyrogenic exotoxin genes, macrolide resistance genes, and emm types were also evaluated. Overall, about 50% of S. pyogenes isolates carried fibronectin-binding protein genes. The prevalence of the prtF1 gene was significantly higher among isolates from noninvasive infections (71.4%) than among isolates from invasive infections (30.8%; P = 0.0037). Strains possessing both the prtF1 and prtF2 genes were more likely to be isolates from noninvasive infections than isolates from invasive infections (50.6% vs 15.4%; P = 0.019). S. pyogenes isolates with streptococcus pyrogenic exotoxin genes (speA and speZ) were more common among isolates without fibronectin-binding protein genes. The speC gene was more frequently identified among isolates with fibronectin-binding protein genes (P = 0.05). Strains belonging to emm75 or emm12 types more frequently harbored macrolide resistance genes than other emm types (P = 0.0094 and P = 0.043, respectively). Strains carrying more than one repeat at the RD2 region of the prtF1 gene and the FBRD region of the prtF2 gene were more prevalent among strains with macrolide resistance genes than among strains negative for macrolide resistance genes. These genes (i.e., the prtF1, prtF2, and spe genes) may enable host-bacteria interaction, and internalization in the host cell, but may not enable infection complications such as invasive diseases.
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The aim of this study is to examine the internalization of nontypeable Haemophilus influenzae (NTHi) into human epithelial cells. Bactericidal assay was applied to examine the effects of antibiotics against cell-adherent NTHi using HEp-2 cells. A trans-well chamber assay was applied to examine the internalization and penetration of NTHi using Detroit562 cells. The adherence of NTHi to HEp-2 cells was noted after 2h of incubation. Azithromycin had a strong bactericidal effect against both cell-associated and non-adherent NTHi, while ceftriaxone did not show bactericidal effects on NTHi adhered to the HEp-2 cells. Three (60.0%) out of five NTHi isolates from the nasopharynx of children with intractable acute otitis media (AOM) internalized into and subsequently penetrated through the epithelial cells at various degrees. Azithromycin had a strong bactericidal effect against the cell-internalized NTHi, while ceftriaxone was bactericidal only against extracellular NTHi. The potential of NTHi as the intracellular pathogen may contribute to the persistent existence of this pathogen that result in the prolonged and intractable clinical course of AOM. Azithromycin may be a therapeutically significant antibiotic for patients with prolonged respiratory tract infections due to NTHi.
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We aimed to define role of tonsillar lymphocytes (TL) and immune cross-reactivity between bacterial-HSP65 and human-HSP60 in Pustulosis palmaris et plantaris (PPP), an intractable chronic disease characterized with pustules and cornification of palms and soles. Two sets of crossover trials were designed by employing SCID mice model. In the first trial, mice were transplanted with tonsillar lymphocytes and skin-grafts from PPP patients (TL group). In the second trial, mice were transplanted with tonsillar lymphocytes from PPP patients and injected with recombinant human HSP60. Control groups were designed for each step. Comparisons were performed for immunologic analyses including infiltration of CD4+ lymphocytes in skin-grafts by immunostaining, and levels of anti-HSP65-IgG and cytokines in mice sera by enzyme-linked immunosorbent assay (ELISA). In TL group, infiltration of CD4+ lymphocytes in skin-grafts were significantly higher than mice transplanted with blood lymphocytes (p<0.05), while anti-HSP65-IgG levels in sera showed non-significant tendency to increase in the TL group. CD4+ cells and anti-HSP65-IgG levels were also well-correlated with each other in TL group (p<0.01). Besides, anti-HSP65-IgG levels were significantly correlated with cytokine levels (IL-6, IFN-gamma) in mice sera (p<0.01). We found strong expression of HSP60 in PPP lesions. Finally, HSP60-stimulation in mice transplanted with TL from PPP patients induced significantly higher anti-HSP65-IgG levels in serum compared to control groups including mice without HSP60-stimulation or peripheral blood lymphocytes-transplanted mice or transplanted with TL from control patients (p<0.05). Our results indicate the pathogenic role of TL and immune cross-reaction between human-HSP60 and bacterial-HSP65 in PPP.
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The aim of this study was to develop and evaluate a scoring system for the management of acute pharyngo-tonsillitis. We conducted a prospective study between May 2004 and June 2005. Patients with acute pharyngo-tonsillitis were evaluated for causative pathogens and were assessed clinical symptoms and pharyngo-tonsillar finding by a clinical scoring system. A total 214 adult patients were enrolled in this study. Streptococcus pyogenes were identified at 13.6%. Thirty-one viruses were also identified by PCR. They were adenovirus (4.8%), influenza virus (1.0%), RS virus (6.3%), and human metapneumovirus (2.9%). Numbers of total white blood cells and levels of C-reactive protein showed a significant positive correlation with clinical scores (p<0.001) and were also higher in cases with S. pyogenes. The clinical scores rapidly improved after the antimicrobial treatments in moderate cases and severe cases. The current study strongly suggested that the clinical scoring system reflected disease severity well and would be very useful for evaluating clinical course and decision making for the antimicrobial treatment of acute pharyngo-tonisllitis.
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• More than 20,000 acute respiratory illnesses of children were studied by physicians in private practice to derive a nine-factor scorecard designed to estimate the probability that a given child's respiratory illness is caused by β-hemolytic streptococci. Each factor was assigned a weight on the basis of the percentage of positive cultures from patients exhibiting that factor. The sum of these individual scores gave the total score. The higher a patient's score, the more probable was a streptococcal infection. Thus, depending on the score, a reasonably accurate prediction of the probability of a streptococcal infection can be made. The accuracy of such predictions compared favorably with that of skilled physicians. (Am J Dis Child 131:514-517, 1977)
Article
The usefulness of erythromycin is limited by its poor pharmacokinetic profile which is characterised by low blood levels and poor gastric acid stability. Erythromycin's short half-life means that a four-times daily dosage schedule is required for effective treatment. In comparison, the azalide structure of azithromycin confers a much improved pharmacokinetic profile. The bioavailability of azithromycin is approximately 37% in humans (25% for erythromycin). Serum concentrations decline in a polyphasic manner and the relatively short serum half-life (11-14 hours recorded 8-24 hours after last dose) is an indication of the initial rapid distribution of drug into the tissues. The low serum levels recorded 24 hours or more after the end of administration are thought to reflect the slow release of azithromycin from tissues. Tissue concentrations exceed serum concentrations by as much as 100-fold following a single 500 mg oral dose. Macrophages and polymorphonuclear leucocytes concentrate azithromycin at levels greater than those found in tissues themselves. During multiple dosing, tissue half-life increases with duration of administration and the tissue to serum ratio further increases. High concentrations of drug are found in tissues such as tonsil, lung, prostate, liver and lymph nodes with relatively low concentrations in fat and muscle. Significantly, the sustained high levels of drug in the tissues appears to correlate with good in vivo activity. Two 1.5 g regimens have been investigated in clinical trials: 500 mg on day 1, followed by 250 mg daily on days 2 to 5; or 500 mg daily for three days.(ABSTRACT TRUNCATED AT 250 WORDS)
Article
Haemophilus influenzae type b is a common cause of systemic bacterial disease in children, and the serotype b capsule is a major determinant of virulence. Nevertheless, as a consequence of the genetic configuration of the capb locus, type b strains become capsule deficient at a high frequency. To investigate the potential biological relevance of the predisposition to capsule loss, we compared the adherent and invasive abilities of several strains of H. influenzae type b and their isogenic capsule-deficient mutants by using cultured human epithelial cells. In all cases the capsule-deficient mutant demonstrated significantly greater adherence and invasion than the encapsulated parent. Transformation of one capsule-deficient mutant to restore encapsulation resulted in a marked decrease in adherence and invasion. All strains were capable of adherence and invasion by a pilus-independent mechanism. We conclude that capsule loss by H. influenzae type b results in enhanced in vitro adherence and invasion, properties that may be relevant to colonization of the nasopharynx and persistence within the respiratory tract. These observations suggest an explanation for the evolution of the capb locus as directly repeated segments of DNA with a consequent predisposition to recombination resulting in capsule loss.
Article
The nasopharyngeal flora of healthy children were compared with flora in children with otitis media caused by nontypable Haemophilus influenzae, Streptococcus pneumoniae and Moraxella catarrhalis. Forty healthy children were followed prospectively and compared with 70 children with 43 episodes of nontypable H. influenzae, 21 episodes of S. pneumoniae and 28 episodes of M. catarrhalis otitis media. Carriage of nontypable H. influenzae (95% vs. 65%, P less than 0.001), S. pneumoniae (91% vs. 52%, P less than 0.005) and M. catarrhalis (86% vs. 52%, P less than 0.001) increased significantly during episodes of otitis media compared with healthy periods. The quantity of nontypable H. influenzae, S. pneumoniae and M. catarrhalis in nasopharyngeal secretions also increased during active infection compared with healthy periods: 3.0 vs. 2.0, P less than 0.005; 3.2 vs. 2.1, P less than 0.001; and 3.3 vs. 2.5, P less than 0.01, respectively. At the same time, nonpathogens of the resident flora, in particular viridans streptococci, declined in carriage: 65% vs. 22%, P less than 0.001. These data suggest that respiratory pathogens become relatively more important in the microenvironment of the nasopharynx during episodes of otitis media. Furthermore the absence of a middle ear pathogen in a nasopharyngeal culture strongly suggests that the pathogen is not present in the middle ear space (negative predictive value greater than 0.96).
Article
Haemophilus influenzae is a common commensal organism of the human respiratory tract that initiates infection by colonizing the nasopharyngeal epithelium. In some individuals, colonization is followed by localized respiratory tract or systemic disease. To gain insight into the mechanisms by which H. influenzae attaches to and persists within the nasopharynx, we examined the interactions between a nonpiliated clinical isolate of H. influenzae and human epithelial cells. We noted substantial adherence that occurred independently of pili and required viable bacteria capable of de novo protein synthesis. Comparison of profiles of outer membrane proteins synthesized during incubation with epithelial cells for adherent and nonadherent bacteria identified several candidate adhesin molecules. In addition, a small number of adherent bacteria were capable of entering epithelial cells in a process that was inhibited by cytochalasin D and colchicine. The suggestion from our studies is that one or more of several newly synthesized nonpilus bacterial proteins are required for maximal in vitro adherence and invasion. We speculate that H. influenzae entry into epithelial cells may provide a mechanism for evasion of host defenses, thereby allowing persistence in the nasopharynx.
Titers of anti-keratin antibodies (AKA) involving IgG and IgM fractions were determined by a solid-phase enzyme-linked immunsorbent assay (ELISA) and were found to be significantly higher on average in patients with pustulosis palmaris et plantaris (PPP) than in normal controls. Positive rates of IgG-AKA and IgM-AKA in sera of PPP patients were 50% and 66%, respectively. Following tonsillectomies, patients with high IgM-AKA titers that decreased after surgery were found to benefit by a marked improvement in their skin lesions. There was a close relationship between the degree of this improvement in the skin lesions and the changes in IgM-AKA titers after the operation. These results suggest that PPP may be a tonsil-related immunological skin disease.
Article
The homologue of groESL from Porphyromonas gingivalis was cloned and sequenced. Nucleotide sequencing suggested an operon containing two open reading frames (ORFs) homologous to groESL operon of Escherichia coli. The upstream ORF consisted of 267 bp corresponding to 89 amino acid residues. The downstream ORF consisted of 1635 bp corresponding to 545 amino acid residues.
Article
Nontypeable Haemophilus influenzae has emerged as an important human pathogen and is associated with a wide range of diseases. As methods for subtyping isolates have improved, our understanding of the epidemiology and pathogenesis of nontypeable H. influenzae disease has grown. These advances have facilitated identification of a number of bacterial factors with potential as vaccine components. In this review we discuss methods of classifying nontypeable H. influenzae, the epidemiology of disease due to these organisms, bacterial and host factors contributing to disease pathogenesis, and the status of efforts to develop an effective vaccine.
Article
In order to correlate the immunomodulatory roles of homologous heat shock proteins with Mr 65 kD (HSP65) to skin diseases, antibody level to recombinant-HSP65 of Mycobacterium leprae was quantified with enzyme-linked immunosorbent assay (ELISA) in the sera of patients. In psoriasis, an insignificant increase was observed in anti-HSP65 IgG (0.111 +/- 0.053, mean +/- SD in 0D492 nm, n = 22), compared with a normal group (0.080 +/- 0.032, n = 9). However, psoriasis of acute guttate-type (PGA), which is often induced after tonsillar infection, showed a significant increase (0.178 +/- 0.032 n = 4, p <0.001), but psoriasis vulgaris did not (PV) (0.101 +/- 0.053, n = 12), nor generalized psoriasis pustulosa (PP) (0.087 +/- 0.025, n = 6). Similarly, patients with palmoplantar pustulosis (PPP) with tonsillar or periodontal infection showed significantly high anti-H5P65 IgG (0.230 +/- 0.065, n = 7, p <0.0001), compared with only a mild increase in PPP without suspected infectious foci (0.139 +/- 0.066, n = 13, p <0.05). Possible staphylococcal infection in the oral cavity was suggested by an additional ELISA assay to staphylococcal antigen: anti-staphylokinase IgG showed a significant increase in PPP with infectious foci (0.110 +/- 0.028 n = 3, p <0.01) compared with the normal group (0.039 +/- 0.014), while PPP without them showed only a mild change (0.060 +/- 0.017, n = 6, p <0.05). We assume that immunoreaction to H5P65 may be involved in psoriatic skin inflammation associated with focal infection.
Article
Pustulosis palmaris et plantaris (PPP) has been considered as one of the typical tonsillar focal infections, based on the marked clinical improvement of the skin lesions after tonsillectomy. In early-stage of PPP, it has been reported that lymphocytes, predominantly CD4-positive cells, infiltrate the palmar and plantar skin. However, the origin and mechanism of infiltration by these lymphocytes is not clear and there are very few reports on whether tonsillar cells react directly with the skin, possibly due to the difficulty of establishing adequate animal models. In this study, we established an experimental animal model of mice with severe combined immunodeficiency (SCID) and observed the reactions of the tonsillar lymphocytes (TL) or peripheral blood lymphocytes (PBL) of patients with PPP to their own plantar skin by transplanting the lymphocytes and skin of the patient into the mice. The results revealed that TL infiltrated the skin to a significantly greater extent than PBL. Most of these lymphocytes were T lymphocytes, and no B lymphocytes were detected in the transplanted skin. Strong expression of LFA-1 and ICAM-1 was observed in the skin after the TL transplantation. These results suggest that TL have a strong affinity for the skin in PPP, and that the adhesion molecules may play an important role in the infiltration by lymphocytes of the skin.
Article
Relationships between the onset of pustulosis palmaris et plantaris, periodontitis and heat shock proteins were studied by using enzyme-linked immunosorbent assay to examine levels of immunoglobulin G (IgG) against Escherichia coli GroEL, a recombinant DnaJ of Actinobacillus actinomycetemcomitans heat shock protein, a synthetic peptide made from the 180th to the 188th amino acids of Mycobacterium bovis BCG Hsp65, and a recombinant human Hsp60, in sera obtained from 43 pustulosis palmaris et plantaris patients judged to have chronic infectious diseases of the oral cavity. We found that the titers of IgG against E. coli GroEL and A. actinomycetemcomitans DnaJ in the sera from pustulosis palmaris et plantaris patients were significantly higher than those in the control group, whereas the titers of IgG against the synthetic M. bovis Hsp65 and the recombinant Hsp60 did not differ significantly. Periodontal therapy and extraction of teeth with periapical infectious resulted in remission of pustulosis palmaris et plantaris and a statistically significant reduction in the levels of IgG against E. coli GroEL in 9 of the 22 patients (41%) examined. We also found that the IgG levels against A. actinomycetemcomitans DnaJ in 6 serum samples of 16 (37%) were reduced, but not significantly, after the treatment. These results suggest that the IgG responses to heat shock proteins partially induced by oral bacteria may be related to the onset of pustulosis palmaris et plantaris in some patients.
Article
Previous reports showed that nontypeable Haemophilus influenzae (NTHi) reside in macrophage-like cells in human adenoid tissue. This study investigated the ability of nonopsonized NTHi and encapsulated H. influenzae type b (Hib) to enter human monocytic and epithelial cells. The number of intracellular bacteria was determined by a viability assay and flow cytometry. To characterize the mechanisms responsible for the internalization of NTHi, different inhibitors of surface molecules, receptor turnover, and the cytoskeleton were used. Hib were found in monocytic cells at very low numbers (<100 bacteria/2×105 cells). In contrast, a great variation in intracellular numbers was detected between the different NTHi isolates (range, 0.0007%–0.28% of the inoculum for monocytes and 0.053%–3.5% for epithelial cells). NTHi entered human monocytic and epithelial cells via a receptor-mediated endocytosis involving mainly a β-glucan receptor that could be blocked by laminarin
Article
Non-typeable Haemophilus influenzae (NTHi) invades host cells by binding of the platelet-activating factor (PAF) receptor via lipooligosaccharide (LOS) glycoforms containing phosphorylcholine (ChoP). The effect of NTHi infection on host cell signalling and its role in NTHi invasion was examined. The infection of human bronchial epithelial cells with NTHi 2019 increased cytosolic Ca2+ levels, and the invasion of bronchial cells by NTHi 2019 was inhibited by pretreatment with the cell-permeant intracellular Ca2+ chelator BAPTA-AM (P = 0.022) or thapsigargin (P = 0.016). Cytosolic inositol phosphate (IP) levels were also increased after infection with NTHi 2019 (P < 0.001), but not after infection with isogenic mutants expressing altered LOS glycoforms lacking ChoP. PAF receptor antagonist reduced NTHi 2019-stimulated IP production in a dose-dependent manner. NTHi 2019 invasion was inhibited by pertussis toxin (PTX) and the phosphatidylinositol-3-kinase inhibitors wortmannin and LY294002. The less invasive strain NTHi 7502 also initiated IP production, but was unaffected by PAF receptor antagonist or PTX. These data demonstrate that the binding of the PAF receptor by NTHi initiates receptor coupling to a PTX-sensitive heterotrimeric G protein complex, resulting in a multifactorial host cell signal cascade and bacterial invasion. Moreover, the data suggest that NTHi strains initiate cell signalling and invade by different mechanisms, and that invasion mediated by PAF receptor activation is more efficient than macropinocytosis.
Article
Pustulosis palmaris et plantaris (PPP) has been considered as one of the typical tonsillar focal infections, based on the marked clinical improvement of the skin lesions after tonsillectomy. Despite the accumulation of data showing the clinical efficacy of tonsillectomy for this skin lesion, fundamental etiological and pathophysiological issues have yet to be addressed. One primary obstacle hindering investigators has been the lack of an appropriate animal model for this human skin disorder. In the early stage of PPP, it has been reported that lymphocytes, predominantly CD4+ T lymphocytes, infiltrate the palmar and plantar skins. However, the origin and mechanism of infiltration by these lymphocytes is not clear and there are very few reports on whether tonsillar mononuclear cells react directly with the skin. We have been intrigued by the ability to engraft human cells onto severe combined immunodeficiency (SCID) mice, together with the opportunity for long-term graft survival and ability to adoptively transfer various human immunocompetent cells. In this review, we addressed the existing deficiencies in our understanding of the relationship between tonsils and PPP by using emerging transplantation technology involving SCID mice.
Article
Pustulosis palmaris et plantaris (PPP) is known to be a skin disease related to tonsillitis, because the pustulosis often become exacerbated during acute tonsillitis and disappears after tonsillectomy. However, etiology of PPP remains unclear. In this study, we investigated the activation of tonsillar T-cell from PPP patients. Furthermore, we analyzed expressions of cytotoxic T-lymphocyte antigen-4 (CTLA4) that is a co-stimulatory molecule for inhibition of T-cell activation and of Smad7 that is a regulatory factor of TGF-beta intracellular signaling. For 47 Japanese patients with PPP who had tonsillectomy, the skin lesion was improved in 87% of PPP patient at 12 months after tonsillectomy. In quantitative immunohistologic analysis, T-cell nodules on tonsillar tissues from PPP patients were more expanded than those from the patients with obstructive sleep apnea syndrome (OSAS) (P = 0.015), and there was a positive correlation between the enlargement and clinical improvement (r = 0.422, P = 0.021). Flow cytometric analysis showed that the numbers of CD4+CD25+ and CD4+CD29+ cells in tonsils from PPP patients increased significantly compared to those from OSAS patients (P = 0.017, P = 0.016, respectively). Using reverse transcription-polymerase chain reaction (RT-PCR) and Western blotting analyses with CD3+ tonsillar lymphocytes, we found that both expressions of Smad7 mRNA and protein were enhanced in PPP patients compared with OSAS patients (P = 0.03, P = 0.02, respectively), but expression of TGF-beta mRNA was not different between 2 groups. Although mRNA expression of CTLA4 was reduced in PPP patients compared with OSAS patients (P = 0.04), the CTLA4 surface protein expression was not different between 2 groups. These data suggest that helper T-cells are frequently activated in tonsils from PPP patients, and this activation may be related to unresponsiveness of TGF-beta1 by overexpression of Smad7. Such hyper-activation of T-cell may increase the risk of elicitation of self-reactive T-cell, being associated with pathogenesis of PPP.