Importance of biomarkers in streptococcal acute tonsillitis & peritonsillar abscess.

Background & objectives Acute tonsillitis is a disease that can often be cured with medical treatment. However, complications may occur during this disease process. One of these complications is peritonsillar abscess. In recent years, biomarkers have been frequently used in the diagnosis of diseases. The aim of the study was to reveal whether peritonsillar abscess develops after acute tonsillitis, and acute tonsillitis can be differentiated using biomarkers and which biomarker has higher predictive value for this differentiation. Methods The control group consisted of individuals who were operated for septoplasty in the otolaryngology clinic, and the acute tonsillitis group consisted of individuals diagnosed with acute tonsillitis in the same clinic. Both groups were statistically compared in terms of mean platelet volume (MPV), neutrophil to lymphocyte ratio (NLR), platelet to lymphocyte ratio (PLR), systemic immune inflammation index (SII), infection discrimination index (IDI), plateletcrit (PCT) and lymphocyte to monocyte ratio (LMR) biomarkers. Statistically, significant biomarker values were compared between the subgroups in the tonsillitis group of those who had only acute tonsillitis and those who had peritonsillar abscess due to acute tonsillitis. Receiver operating characteristics (ROC) curve analysis was performed on biomarkers for their ability to predict the presence of peritonsillar abscess. Results When the individuals who had only acute tonsillitis and those who had acute tonsillitis with peritonsillar abscess were compared in terms of biomarkers, there was a statistically significant difference between the mean MPV, SII and PCT (P=0.010, 0.021, 0.023, respectively). ROC analysis was performed to calculate the sensitivity and specificity of MPV, PCT and SII for the diagnosis of acute tonsillitis with peritonsillar abscess (sensitivity-specificity for MPV 51.9-72.7%, for SII 94.2-32.7%, for PCT 71.2-50.9%, respectively). Interpretation & conclusions MPV, SII and PCT biomarkers may be useful to help clinicians predict peritonsillar abscess due to acute tonsillitis.

Extranodal natural killer/T-cell lymphoma with tonsil involvement: a case report.

BACKGROUND: Extranodal natural killer/T-cell lymphoma (ENKTL) with tonsil involvement is not common, especially in children. CASE PRESENTATION: A 13-year-old girl presented with an unexplained sore throat for more than 2 months, together with intermittent fever and suppurative tonsilitis. Nasopharyngoscopy revealed a pharyngeal mass. Enhanced computed tomography (CT) scan showed tonsillar hypertrophy and punctate calcification. Chronic pyogenic granulomatous inflammation with pseudoepithelial squamous epithelial hyperplasia was observed in left tonsil, and pyogenic granulomatous inflammation and a small number of T-lymphoid cells were detected in the right tonsil. The immunohistochemical results showed CD2+, CD3+, CD4+, CD5+, CD8+, granzyme B+, and TIA-1+. The Ki-67 proliferation index was 20%. The case showed T cell receptor gene rearrangement. Finally, the case was diagnosed as ENKTL of stage II with tonsil involvement. The patient received 6 cycles of chemotherapy with SMILE regimen, and showed complete response with no recurrence in the follow-up. CONCLUSION: We presented a rare case of ENKTL with tonsil involvement in a child. The patient showed complete response to the SMILE chemotherapy with no recurrence.

Histological Correlation between Tonsillar and Glomerular Lesions in Patients with IgA Nephropathy Justifying Tonsillectomy: A Retrospective Cohort Study.

Tonsillectomy with steroid pulse therapy (SPT) has been established as an effective treatment for immunoglobulin A nephropathy (IgAN) in Japan. However, the underlying mechanisms supporting tonsillectomy remain unclear. This study assessed palatine tonsils from 77 patients with IgAN, including 14 and 63 who received SPT before and after tonsillectomy, respectively. Tonsils from 21 patients with chronic tonsillitis were analyzed as controls. Specific tonsillar lesions were confirmed in patients with IgAN, correlating with active or chronic renal glomerular lesions and SPT. T-nodule and involution of lymphoepithelial symbiosis scores in tonsils correlated with the incidence of active crescents and segmental sclerosis in the glomeruli, respectively. The study revealed an essential role of the tonsil-glomerular axis in early active and late chronic phases. Moreover, the SPT-preceding group demonstrated no changes in the T-nodule score, which correlated with active crescent formation, but exhibited a considerable shrinkage of lymphatic follicles that produced aberrant IgA1. The study underscores the involvement of innate and cellular immunity in IgAN and advocates for tonsillectomy as a necessary treatment alongside SPT for IgAN, based on a stepwise process.

Accelerated involution of germinal center in palatine tonsils in IgA nephropathy.

BACKGROUND: Altered immunological responses in the palatine tonsils may be involved in the pathogenesis of IgA nephropathy (IgAN). The germinal center serves as the site for antigen-specific humoral immune responses in the palatine tonsils. Germinal center involution is frequently observed in the palatine tonsils of IgAN (IgAN tonsils). However, the pathogenic significance of these characteristic changes remains unclear. This study aimed to investigate the morphological changes in secondary lymphoid follicles in IgAN tonsils and to evaluate the correlation between the morphometric results and the clinicopathological severity of IgAN. METHODS: The tonsils of age-matched patients with recurrent tonsillitis (RT tonsils) were used as controls. The correlation between the degree of lymphoid follicular involution and histopathological severities in clinical or kidney biopsy was evaluated. RESULTS: In total, 87 patients with IgAN were included (48% male, median age 35 years, median estimated glomerular filtration rate: 74 mL/min/1.73 m2). Compared to RT tonsils, IgAN tonsils showed smaller median sizes of lymphoid follicles and germinal centers (P < 0.001). The relative areas of lymphoid follicles (%LFA) and germinal centers (%GCA) in the total tonsillar tissue were smaller in the IgAN tonsils than in the RT tonsils (P < 0.001). In contrast, the median proportion of mantle zones in the total tonsillar tissue was comparable between the groups. A lower %LFA was associated with a longer period from the onset of urinary abnormalities to biopsy diagnosis and higher urinary protein excretion (P = 0.01). %LFA showed significant negative correlations with frequencies of glomeruli with both global and segmental sclerosis. CONCLUSIONS: The present study confirmed accelerated germinal center involution in the tonsils of patients with IgAN. This characteristic change in the IgAN tonsil correlates with heavy proteinuria and advanced chronic histopathological changes in the kidneys, thereby suggesting the involvement of repeated tonsillar immunoreactions during IgAN progression.

Paired qualitative and quantitative analysis of bacterial microcolonies in the tonsils of patients with tonsillar hyperplasia.

The discovery of bacterial microcolonies in tonsillar tissue of patients with tonsillar hyperplasia has raised the question of their role in provoking the local immune response. Tonsils collected from patients undergoing tonsillectomy were stained for three clinically relevant bacterial taxa and lymphocytes. The bacterial composition and abundance of microcolonies was investigated using a combination of laser-microdissection, amplicon sequencing and Droplet Digital polymerase chain reaction. Microcolonies were detected in most samples (32/35) with a high prevalence of Haemophilus influenzae (78% of samples). B and T cell lymphocytes were significantly higher in the epithelium adjacent to microcolonies compared to epithelium distal to microcolonies. Furthermore, significant positive and negative correlations were identified between bacterial taxa and lymphocytes. Genus Streptococcus, which includes Group A Streptococcus (traditionally described as the main pathogen of tonsillar hyperplasia), was found in low abundance in this study. These results suggest other potential pathogens may be involved in stimulating the local immune response leading to tonsillar hyperplasia.

IL-1R8 expression in DLBCL regulates NK cell recruitment and influences patient prognosis.

The precise biological function of Interleukin-1 receptor 8 (IL-1R8) in diffuse large B-cell lymphoma (DLBCL) is still not well understood. Our goal is to decipher the profile of IL-1R8 expression status in DLBCL and to explore how IL-1R8 is involved in DLBCL progression. Utilizing a tissue microarray consisting of 70 samples of DLBCL tumors alongside 15 samples of tonsillitis, our investigation revealed a parallel expression profile of IL-1R8 between the tumor tissues and tonsillitis samples (p > 0.05). Nevertheless, an intriguing association emerged, as heightened expression of IL-1R8 correlated significantly with unfavorable survival outcomes in patients with DLBCL (p < 0.05). The status of IL-1R8 expression did not directly regulate proliferation (p > 0.05) and apoptosis (p > 0.05) in DLBCL cells via CCK8 and apoptotic assays. Subsequent chemotaxis analysis indicated that natural killer (NK) cell recruitment could be suppressed by IL-1R8 signaling in DLBCL, at least partially through CXCL1 inhibition (p < 0.05). The status of IL-1R8 expression in tumor tissues exhibited a negative correlation with the density of CD57+ NK cell infiltration (p < 0.05), while it did not demonstrate a significant association with CD3+ T cells (p > 0.05), CD68+ macrophages (p > 0.05), or S-100+ dendritic cells (p > 0.05). In line with this observation, elevated levels of NK cell infiltration demonstrated a significant positive correlation with improved overall survival (OS) among patients diagnosed with DLBCL (p < 0.05). Our data suggests the immuno-regulating potential of IL-1R8 through NK cell recruitment in DLBCL, providing novel insights into future immuno-modulating therapies.

Comparison of innate lymphoid cells from tissue and blood in chronic tonsillitis and tonsillar hypertrophy.

OBJECT: Recurrent tonsillitis and tonsillar hypertrophy are two common diseases in children for which tonsillectomy is the definitive solution. The underlying causes of both diseases are not fully known. The aim of this study was to identify the predominant innate lymphoid cells in both diseases of the palatine tonsils, which are known to play an important role in the immune system. METHODS: Children who underwent tonsillectomy were divided into two groups as recurrent tonsillitis and tonsillar hypertrophy according to the indication for surgery. The proportions of innate lymphoid cell (ILC) groups and IFN-gamma, IL-10 and IL-17 secreting T lymphocyte cells were determined in tonsil and blood samples obtained during surgery. Local and peripheral immune responses were evaluated. Innate immune responses and acquired immune responses were compared. RESULTS: The results of our study showed that the proportions of the innate lymphoid cell 1 group (ILC1) were similar in tonsil tissue in patients with recurrent tonsillitis and tonsil hypertrophy, with no statistically significant difference. It was observed that the innate lymphoid cell 2 group (ILC2) was the predominant group in tonsil hypertrophy, the innate lymphoid cell 3 group (ILC3) was the predominant innate lymphoid cell group in recurrent tonsillitis, and the proportion of IL-17 secreting T lymphocytes in blood and tonsillar mononuclear cells was higher in recurrent tonsillitis patients than in tonsil hypertrophy patients. CONCLUSION: With the results obtained, the predominant innate lymphoid cells in the pathogenesis of both diseases were identified and local and peripheral responses were compared. These findings may be a guide for possible medical treatments for both diseases in the future.

Incidental histopathological diagnosis of tonsilar actinomycosis: A report of four cases.

Actinomycosis of the tonsils is uncommon even though the causative organisms are normal commensal of the oropharyngeal tract that may colonise the tonsillar crypts with resultant infection in apparently healthy individuals. Diagnosis is often incidental in tonsillectomy specimens sent to the pathology laboratory for varied diseases. This is a 10-year study of tonsillectomy specimens diagnosed with actinomycosis. Specimens were formalin-fixed and paraffin processed and stained with haematoxylin and eosin, gromott methenamine silver and periodic acid-Schiff. Four cases of tonsillar actinomycosis were diagnosed from a total of 772 tonsillectomy specimens. Histologically, characteristic oeosinophilic granules with peripheral radial protuberances surrounded by microabscesses were seen. Tonsillar actinomycosis is often an incidental diagnosis; however, a high index of suspicion should be entertained in patients with recurrent tonsillitis and/or tonsillar hypertrophy of unknown cause.

Hamartomatous polyp of the palatine tonsil: histological considerations and review of the literature.

The hamartomatous polyp is a rare benign hamartoma of the palatine tonsil, usually encountered during the second decade of life. It may be reported under various terms in the literature, like lymphangioma of the tonsil, angiofibrolipoma, lymphangiomatous tonsillar polyp and lymphangiectatic fibrous polyp. Macroscopically, it appears as a large, pale, pedunculated mass. Typically, a hamartomatous polyp is asymptomatic or manifests mild symptoms, like foreign body sensation. It is not related to a generalised lymphatic malformation process. Despite its typical appearance, an excisional biopsy is necessary to rule out a malignancy. Histological findings are consistent with a squamous epithelial covering, a core of loose fibrous and adipose tissue with sparse lymphoid aggregations and dilated lymphatic channels filled with lymph and lymphocytes. Several embryologically based theories suggested its pathogenesis; however, recurrent tonsillitis does not play an established role. A typical tonsillectomy is suggested as a sufficient therapeutical approach with no tendency for recurrence.

Current bacteriological profile of tonsillar surface versus core tissue in patients undergoing tonsillectomy.

PURPOSE: The current study explored the bacteriological profile in the tonsillar core in patients undergoing tonsillectomy and compared it with tonsillar surface organisms. The antibiotic sensitivity and resistance patterns were also studied. MATERIALS AND METHODS: This prospective and observational study was conducted in the Department of Otorhinolaryngology, Head and Neck surgery in a tertiary care institute, on a total of sixty-one patients clinically diagnosed as chronic tonsillitis and undergoing tonsillectomy. Tonsillar surface culture swabs and core tissue specimen were sent in separate vials. The culture isolates and their antibiotic sensitivity were analysed and compared. RESULTS: Bacterial growth was detected in 61% of the cases, in cultures from the surface or the core tissue or both. 50.8% specimens of core tissue versus 31.1% of tonsillar surface samples revealed bacterial growth. Of the 13 cases in which surface and core both had bacterial culture growth, the same pathogens were identified in 8 cases. Pseudomonas aeruginosa was the most commonly reported organism in both tonsillar core as well as surface swab cultures. CONCLUSION: Our study shows that surface swab culture is not always a reliable indicator of organisms present in core of tonsils in cases of chronic infection. The surface swabs did not always match the pathogens in the core. The higher prevalence and the variability of pathogenic organisms in the core tissue as compared to the surface suggests that a targeted antibiotic treatment based on surface culture swabs is unlikely to eradicate them.

[Clinical and morphological picture of chronic tonsillitis and its pathogenetic relationship with periodontal diseases: literature review]. / Kliniko-morfologicheskaya kartina khronicheskogo tonzillita i ego patogeneticheskaya svyaz' s parodontopatiyami: obzor literatury.

The article is devoted to the study of chronic tonsillitis from the point of view of the clinical and morphological picture of the disease, histological changes in the palatine tonsils in various forms of CT. The authors raise the question of studying the assessment of the effect of conservative therapy on the morpho-functional state of the palatine tonsils. The article focuses on the study and influence of extratonsillary oropharyngeal loci of focal infection, such as periodontopathies, on the course of chronic inflammation of the palatine tonsils. The association of periodontitis with CT and various somatic diseases has been demonstrated. The necessity of further study of the cause-and-effect relationships of CT and periodontal diseases, as well as a more in-depth study of the morphological picture in CT using immunohistochemical methods of research, which will allow us to develop new principles for the diagnosis and treatment of this disease, is justified.

FEATURES OF THE CONNECTIVE TISSUE COMPONENT OF THE PALATINE TONSILS IN PATIENTS WITH RECURRENT TONSILLITIS.

OBJECTIVE: The aim: To explore the morphological changes of palatine tonsil at the levels of the epithelial layer and connective tissue; to determine the relative area of the connective tissue component in the tonsillar tissue (fibrosis) in patients with recurrent tonsillitis compared to the control. PATIENTS AND METHODS: Materials and methods: This study presents a morphological assessment of the palatine tonsils of 10 people. Tonsils' material with surrounding tissue was fixed in 10% formalin solution. The samples were dehydrated in increasing ethanol concentrations, cleared in xylol, impregnated with paraffin. Microscopy was then performed with samples stained beforehand. RESULTS: Results: In the samples of patients with recurrent tonsillitis pericapsular sclerosis was noted, along with thickening of interlobular septa and pronounced subepithelial fibrosis. A ratio of the dense connective tissue surface area to the total surface area of tonsil tissue was determined. The control group showed a statistically significant decrease in the degree of sclerosis of the tonsil stroma. CONCLUSION: Conclusions: Multiple changes were found in the tonsils of patients with recurrent tonsillitis at the level of the epithelial layer that manifested in structural alterations. Significant and irreversible changes were also observed in the connective stroma of the tonsil - pericapsular sclerosis, thickening of interlobular septa, and pronounced subepithelial fibrosis. A statistically significant increase in the relative surface area of the connective tissue component of the tonsil (fibrosis) by a factor of 1,26 was noted in patients with recurrent tonsillitis compared to the results of the control group of patients.

Fluorescence in situ hybridisation in Carnoy's fixed tonsil tissue.

Fluorescence in situ hybridisation (FISH) is a powerful molecular technique that enables direct visualisation of specific bacterial species. Few studies have established FISH protocols for tonsil tissue in Carnoy's fixative, accordingly limiting its application to investigate the pathogenesis of tonsillar hyperplasia. Tonsil tissue from 24 children undergoing tonsillectomy for either recurrent tonsillitis or sleep-disordered breathing were obtained during a previous study. The specificity of each of the five FISH probes (Fusobacterium spp., Bacteroides spp., Streptococcus spp., Haemophilus influenzae and Pseudomonas spp.) were successfully optimised using pure and mixed bacterial isolates, and in Carnoy's fixed tonsil tissue. Bacteroides spp. were present in 100% of patients with microcolonies. In comparison, the prevalence of Fusobacterium spp. was 93.8%, Streptococcus spp. 85.7%, H. influenzae 82.35% and Pseudomonas spp. 76.5%. Notable differences in the organisation of bacterial taxa within a single microcolony were also observed. This is the first study to establish a robust FISH protocol identifying multiple aerobic and anaerobic bacteria in Carnoy's fixed tonsil tissue. This protocol provides a strong foundation for combining histological and microbiological analyses of Carnoy's fixed tonsil samples. It may also have important implications on the analysis of microorganisms in other human tissues prepared using the same techniques.

Do tonsils regrow after partial tonsillectomy? - Histology of regrown tonsils and predisposing factors for tonsillar regrowth.

OBJECTIVES: It has been suggested that after partial tonsillectomy, referred here as tonsillotomy (TT), the remaining tonsillar tissue might be altered, leading to scarring or chronic tonsillitis. The objectives were to compare the histology of regrown tonsillar tissues with native tonsils and to assess the incidence of reoperations and predictive factors for tonsillar regrowth after TT. METHODS: Tonsillar tissues of 1) children that underwent TT and later requiring resurgery and 2) children operated on for the first time with TT were prospectively analysed. To assess the incidence of resurgery because of tonsillar regrowth and predictive factors for tonsillar regrowth, the data covering TTs and tonsillectomies performed in 2009-2020 were retrospectively retrieved. RESULTS: Altogether 11 children formed the regrowth group, with the control group consisting of 19 children. In the histological analysis of tonsillar tissues, neutrophil infiltration in the epithelium and crypts and severe lymphoplasmacytic infiltration in the epithelium were significantly more frequent in samples of the regrowth group relative to the control group. The number of germinal centres was greater in the regrowth group. In the retrospective analysis of 3141 children, the incidence of resurgery after primary TT was 1.9%. Logistic regression analysis showed that age was the only significant predisposing factor for resurgery. CONCLUSION: Inflammatory cells were present more often in regrown tonsillar tissues but there was no evidence of severe chronic inflammation or scarring in the regrown tonsils. The risk of resurgery after TT was low. Young age predisposed to tonsillar regrowth, no other risk factors were found.

Histopathological role of vitamin D deficiency in recurrent/chronic tonsillitis pathogenesis: Vascular epithelial growth factor-mediated angiogenesis in tonsil.

OBJECTIVES: Our aim in this study is to reveal the role of vitamin D deficiency in the pathogenesis of recurrent/chronic tonsillitis and to determine the expression of vascular epithelial growth factor (VEGF). MATERIAL AND METHODS: This study was conducted between September and February. Thirty-two patients between the ages of 3 and 35 (mean age 9.71) with recurrent episodes of chronic tonsillitis were selected. Patients were divided into four groups according to their 25OHD levels. Patients with 25OHD levels 0-10 ng/ml were determined as Group 1, 11-20 ng/ml Group 2, 21-30 ng/ml Group 3, and 31-50 ng/ml control Group 4. Routine histological tissue sampling was performed for histopathological evaluation of the tonsillar tissues under light microscope (LM). Five micron sections were taken from the paraffin blocks and stained with Hematoxylin Eosin (HE) and Trichrome Masson (TCM). VEGF expression was examined immunohistochemically for each group. RESULTS: Our analysis showed VEGF expression in all study groups (32 tonsillar tissues). Group 1 and Group 2 histopathological scores were significantly higher than the other groups (p < .001). There were significant differences in VEGF expressions between the four groups (p < .001). 25OHD levels of the patients in Groups 1 and 2 with strong VEGF expression were significantly lower than the other groups (p < .001). CONCLUSIONS: In conclusion, this study showed an increased angiogenesis in tonsil and an increase in VEGF expression of the tonsillar surface epithelium when blood serum 25OHD levels <20 ng/ml.

Relationship between IgA Nephropathy and Porphyromonas gingivalis; Red Complex of Periodontopathic Bacterial Species.

IgA nephropathy (IgAN) has been considered to have a relationship with infection in the tonsil, because IgAN patients often manifest macro hematuria just after tonsillitis. In terms of oral-area infection, the red complex of periodontal bacteria (Porphyromonas gingivalis (P. gingivalis), Treponema denticol (T. denticola) and Tannerella forsythia (T. forsythia)) is important, but the relationship between these bacteria and IgAN remains unknown. In this study, the prevalence of the red complex of periodontal bacteria in tonsil was compared between IgAN and tonsillitis patients. The pathogenicity of IgAN induced by P. gingivalis was confirmed by the mice model treated with this bacterium. The prevalence of P. gingivalis and T. forsythia in IgAN patients was significantly higher than that in tonsillitis patients (p < 0.001 and p < 0.05, respectively). A total of 92% of tonsillitis patients were free from red complex bacteria, while only 48% of IgAN patients had any of these bacteria. Nasal administration of P. gingivalis in mice caused mesangial proliferation (p < 0.05 at days 28a nd 42; p < 0.01 at days 14 and 56) and IgA deposition (p < 0.001 at day 42 and 56 after administration). Scanning-electron-microscopic observation revealed that a high-density Electron-Dense Deposit was widely distributed in the mesangial region in the mice kidneys treated with P. gingivalis. These findings suggest that P. gingivalis is involved in the pathogenesis of IgAN.

Relationship of endoplasmic reticulum stress with the etiopathogenesis of chronic tonsillitis and tonsillar hypertrophy in pediatric patients: a prospective, parallel-group study.

OBJECTIVES: Tonsil tissue is a very important component of the human immunity system, contributing to the functioning of the cellular and humoral defence system, especially in childhood. The endoplasmic reticulum (ER) is an organelle that has a very important function in the balanced functioning of cells, in which the accumulation of a cellular protein called ER stress occurs in case of dysfunction. ER stress influences the pathogenesis of many diseases and immune system functions. We aimed to investigate the relation between the diseases of tonsil tissue and ER stress response to elucidate the mechanisms of diseases related with the immune system. METHODS: A prospective study was conducted in 46 children aged between 2 and 16 years who underwent tonsillectomy for chronic tonsillitis or tonsillar hypertrophy. Tonsil tissue was separated into two groups according to their size and evaluated in terms of ER stress markers and apoptosis markers by Real-time PCR and Western blot analysis. RESULTS: The ΔCT levels of ER stress markers (ATF4, ATF6, CHOP, GRP78, EIF2AK3, ERN1, GRP94) were greater in children with chronic tonsillitis (p < 0.005). In contrast, the tonsillar hypertrophy group had greater ΔCT levels of apoptosis markers (BAX, BCL-2) according to the Real-time PCR method (p < 0.005). According to the Western blot analysis, the normalized levels of ATF4, ATF6, CHOP, GRP78, and ERN1 genes were found greater in the chronic tonsillitis group than the tonsillar hypertrophy group. There was no difference between the two groups in terms of normalized BCL-2 and BAX levels by Western blot analysis. CONCLUSION: This is the first study in the literature investigating the effect of the ER stress pathway on the etiopathogenesis of tonsil diseases. It was concluded that the ER stress pathway plays a role in the etiopathogenesis of chronic tonsillitis. Investigating the relationship between ER stress and structures such as the tonsil tissue that make up the immune system can help create new treatment strategies. CLINICAL TRIAL REGISTRATION: Trial Registration ClinicalTrials.gov Identifier: NCT04653376.

Interaction between Ras and Bcl2L12 in B cells suppresses IL-10 expression.

The pathogenesis of recurrent tonsillitis is to be further investigated. B cell-derived interleukin (IL)-10 plays a critical role in immune regulation. Ras activation plays an important role in cancer and many immune disorders. This study aims to investigate the role of Ras activation in down regulating IL-10 expression in tonsillar B cells. Surgically removed tonsil tissues were collected from patients with recurrent acute tonsillar inflammation; B cells were isolated from the tonsillar tissues by flow cytometry sorting to be analyzed by the Ras-specific enzyme-linked immunosorbent assay and pertinent immunological approaches. We found that, compared to peripheral B cells (pBC), B cells isolated from the tonsillar tissues with recurrent inflammation (tBC) showed higher Ras activation, lower IL-10 expression and higher Bcl2L12 expression. Bcl2L12 formed a complex with GAP (GTPase activating protein) to prevent Ras from deactivating. The Ras activation triggered the MAPK/Sp1 pathway to promote the Bcl2L12 expression in B cells. Bcl2L12 prevented the IL-10 expression in tBCs, that was counteracted by inhibition of Ras or the Ras signal transduction pathway. In conclusion, Bcl2L12 interacts with Ras activation to compromise immune tolerance in the tonsils by inhibiting the IL-10 expression in tBCs. Inhibition of Bcl2L12 can restore the IL-10 expression in tBCs.

Analysis of microbial changes in the tonsillar formalin-fixed paraffin-embedded tissue of Chinese patients with IgA nephropathy.

BACKGROUND: Immunoglobulin A nephropathy (IgAN) is a prevalent chronic glomerular disease contribution to end-stage renal failure (ESRD). The tonsillar microbiota is closely associated with IgAN diseases based on the mucosal immune response. However, the composition and function of in tonsillar microbiota in participant patients with IgAN remains unknown. In this study, we detected the tonsillar microbiota changes of IgAN patients in Heilongjiang province located in northeast China. MATERIAL AND METHODS: We collected from 21 patients with IgAN and 16 patients with chronic tonsillitis (CT) who had undergone tonsillectomy previously. Histological review of all samples from formalin-fixed paraffin-embedded (FFPE) tissue were performed. Extracted DNA from FFPE tissue blocks, after that V4 regions of 16S ribosomal RNA (rRNA) sequencing and comparative analyses of tonsillar flora between two groups were performed. The statistical analysis used the SPSS version of 21. RESULTS: Visualization of microorganisms by Gram and Warthin-Starry (WS) silver stains, preliminarily observed the morphological characteristics of microbiome in FFPE tissue cases, such as bacteria or fungi. Tonsillar FFPE samples from the IgAN patients and CT controls showed significant differences in tonsillar microbial certain compositions and functions. We found that there were eight dominant genera that can be available to distinguish IgAN patients from CT controls. Compared with CT controls, at genus level, the relative abundances of Methylocaldum and unclassified_f_Prevotellaceae were significantly higher, while the abundances of Anaerosphaera, Halomonas, Trichococcus, Peptostreptococcus, norank_f_Synergistaceae and unclassified_k_norank_d_Bacteria were significantly lower in IgAN patients. Principal co-ordinates analysis (PCOA) distinguished IgAN patients from CT controls, and receiver operating characteristic (ROC) curves analysis confirmed that the diagnosis of disease has certain diagnostic significance. In addition, Functional analysis revealed that partly Enzymes and KOs were increased in the IgAN patients. CONCLUSIONS: Histological screening results were very helpful for further gene sequencing, not only to supplement the observation of bacterial morphology and structure, but also to prepare for subsequent gene sequencing and bioinformatics analysis. We elucidated subtle relevance between changes in tonsillar microbiota and IgAN patients, which can be utilized to predict the incidence of IgAN disease. In addition, we predicted that some enzymes, and KOs were closely related to IgAN.