Cells In Tonsils Research Articles

Effects of plant-derived immunomodulators and lizak preparation in vitro taken separately or in combination on the immune factors of palatine tonsils of children with chronic tonsillitis

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.

Establishment and characterization of the pig tonsil epithelial (PT) cell line as a new model for persist infection of Japanese Encephalitis Virus

Japanese encephalitis virus (JEV) causes a serious zoonotic disease worldwide, pig is the reservoir and amplifying host of JEV. JEV can persist infect tonsil in pig, but the relation between persist infection in tonsil and reservoir are not clear until now. A stable pig tonsil cell line is necessary for JEV persist infection research. In this study, we established a continuous epithelial cell line, named PT cell, from the pig tonsil. This cell is susceptible to JEV. We determined the growth characteristics, molecular properties, microstructure profiles of PT cell. JEV is easy to enter PT cell which may partly explain the reason of persist infection. We further determined that LMAN2L, a mannose lectin proteins, is the primary viral receptors for JEV entry in PT cell. IFITM3, an cellular surface antiviral factor, is underexpression in PT cell after JEV infection. All these results provide solid evidence that PT cell will promote additional research on JEV persist infection in pig tonsil.

CD4+ T Follicular Helper Cells in Human Tonsils and Blood Are Clonally Convergent but Divergent from Non-Tfh CD4+ Cells

T follicular helper (Tfh) cells are fundamental for B cell selection and antibody maturation in germinal centers. Circulating Tfh (cTfh) cells constitute a minor proportion of the CD4+ T cells in peripheral blood, but their clonotypic relationship to Tfh populations resident in lymph nodes and the extent to which they differ from non-Tfh CD4+ cells have been unclear. Using donor-matched blood and tonsil samples, we investigate T cell receptor (TCR) sharing between tonsillar Tfh cells and peripheral Tfh and non-Tfh cell populations. TCR transcript sequencing reveals considerable clonal overlap between peripheral and tonsillar Tfh cell subsets as well as a clear distinction between Tfh and non-Tfh cells. Furthermore, influenza-specific cTfh cell clones derived from blood can be found in the repertoire of tonsillar Tfh cells. Therefore, human blood samples can be used to gain insight into the specificity of Tfh responses occurring in lymphoid tissues, provided that cTfh subsets are studied.

Different doses of tannin reflect a double-edged impact on broiler chicken immunity

Tannin is one of the most common phytochemical secondary phenolic metabolites, which is widely distributed in various plant production systems. Dietary intake of tannin can exert different actions on the immune system. The aim of this study was to evaluate the influence of different concentrations on broiler chicken immunity. Three groups (n = 10 in each group) were evaluated: control group given a normal basal diet, high tannic acid (HT) group given high-dose tannic acid (30 g/kg diet) and low tannic acid (LT) group given low-dose tannic acid (0.5 g/kg diet) for 35 days. Feed intake and body weight were measured weekly. Cells were isolated from thymus, spleen and caecal tonsils at the end of the study. Lymphocyte subsets, monocytes phagocytosis and cytokine mRNA expression in spleen were evaluated. The results showed that HT group chickens had decreased daily gain, final body weight, daily feed intake and relative weights of lymphoid organs compared with other groups. In thymus, spleen and caecal tonsils, relative CD4+, CD8+, CD4+CD8+ and γδ+ cell populations in the HT group were significant decreased in comparison with those of other groups. The relative B cell population in the HT group was also significant decreased. Cytokine mRNA expression in spleen cells of the HT group was also significantly lower than that in other groups. Conversely, CD4+CD8+ subpopulations in spleen and caecal tonsils, γδ+ cells in spleen and B cells in caecal tonsils of the LT group were significantly higher than those of the control group. In addition, IFN-γ mRNA expression in the LT group was significantly higher than that of the control group. These results demonstrated that dietary intake of tannin had positive and negative effects on broiler chicken immunity in a dose-dependent manner.

Poly-functional T helper cells in human tonsillar mononuclear cells.

Tonsils are important lymphoid organs in which B cells and T cells complete their maturation and identify cells that are infected by pathogens. However, the functions of T cells in human tonsils remain unclear, especially the characteristics of polyfunctional CD4+ T helper cells. In this study, we used multi-color flow cytometry to analyze the expression or co-expression of effector cytokines in CD4+ T cells from tonsillar tissues. We have demonstrated that tonsillar CD4+ T cell can express various Th effector cytokines after short-term polyclonal stimulation, and that cytokine-producing CD4+ T cells were CD45RO+ T cells. In addition, we analyzed the co-expression of two or more kinds of cytokines at the level of a single cell. The results showed that tonsillar CD4+ T cells exhibited polyfunctionality by co-expressing two to five kinds of cytokines in the same time. These data furnished a basic theory for further understanding the differentiation of polyfunctional Th cells in human tonsils and their functions in resisting invasive microorganisms.

Immunomodulating properties of Cinnabsin

Introduction: Complex natural medicines are successfully used in various inflammatory diseases of the upper respiratory tract, but their immunomodulatory activity has not been studied enough, in particular in case of Cinnabsin. Therefore, the aim of the study was to determine the effect of the medicine on the production of cellular and humoral factors of immunity and inflammation by tonsil cells of the children. Materials and methods: The studies were performed in vitro using 24-hour cultures of children`s pharyngeal tonsil cells after adenotomy. Altogether 90 samples of cultures were examined before and after the influence of the medicine. The effect of complex natural medicine Cinnabsin on the cells production of various cytokine groups (γ-interferon, interleukins 1, 4, 10), expression of surface antigens CD14+, CD16+, and cytolytic activity of natural killer cells were investigated. Methods of enzyme immunoassay, light microscopy and cellular cytolysis with xenogenic targets were used. Statistical processing was carried out with the Student`s non-parametric criterion, and the Fisher`s angle transformation method when processing ratios. Results: Cinnabsin, both in dosage 10 μg/ml and 100μg/ml, increased the number of cells with the expression of the antigen CD16+ and partially CD14+, activated the natural cytotoxic cells of the tonsils, reduced the production of IL-1β and increased the production of IL-10 and γ-interferon in cell culture. Conclusion: According to the sum of the effects and the immunomodulation vectors, it is possible to state the modulating effect of the medicine on the activity of various immunity reactions, which allows concluding the pronounced anti-inflammatory and antiviral effect of Cinnabsin.

HIV-2 Depletes CD4 T Cells through Pyroptosis despite Vpx-Dependent Degradation of SAMHD1.

Human immunodeficiency virus type 2 (HIV-2) infection results in a milder course of disease and slower progression to AIDS than does HIV-1. We hypothesized that this difference may be due to degradation of the sterile alpha motif and HD domain 1 (SAMHD1) host restriction factor by the HIV-2 Vpx gene product, thereby diminishing abortive infection and pyroptotic cell death within bystander CD4 T cells. We have compared CD4 T cell death in tonsil-derived human lymphoid aggregate cultures (HLACs) infected with wild-type HIV-2, HIV-2 ΔVpx, or HIV-1. In contrast to our hypothesis, HIV-2, HIV-2 ΔVpx, and HIV-1 induced similar levels of bystander CD4 T cell death. In all cases, cell death was blocked by AMD3100, a CXCR4 entry inhibitor, but not by raltegravir, an integrase, indicating that only early life cycle events were required. Cell death was also blocked by a caspase-1 inhibitor, a key enzyme promoting pyroptosis, but not by a caspase-3 inhibitor, an important enzyme in apoptosis. HIV-1-induced abortive infection and pyroptotic cell death were also not reduced by forced encapsidation of HIV-2 Vpx into HIV-1 virions. Together, these findings indicate that HIV-2 and HIV-1 support similar levels of CD4 T cell depletion in vitro despite HIV-2 Vpx-mediated degradation of the SAMHD1 transcription factor. The milder disease course observed with HIV-2 infection likely stems from factors other than abortive infection and caspase-1-dependent pyroptosis in bystander CD4 T cells.IMPORTANCE CD4 T cell depletion during HIV-1 infection involves the demise of bystander CD4 T cells due to abortive infection, viral DNA sensing, inflammasome assembly, and death by caspase-1-dependent pyroptosis. HIV-2 infection is associated with milder disease and lower rates of CD4 T cell loss. We hypothesized that HIV-2 infection produces lower levels of pyroptosis due to the action of its Vpx gene product. Vpx degrades the SAMHD1 restriction factor, potentially reducing abortive forms of infection. However, in tonsil cell cultures, HIV-2, HIV-2 ΔVpx, and HIV-1 induced indistinguishable levels of pyroptosis. Forced encapsidation of Vpx into HIV-1 virions also did not reduce pyroptosis. Thus, SAMHD1 does not appear to play a key role in the induction of bystander cell pyroptosis. Additionally, the milder clinical course of HIV-2-induced disease is apparently not explained by a decrease in this inflammatory form of programmed cell death.

Histological and immunohistochemical studies of the proximal caecum and caecal tonsils of quail (Coturnix coturnix japonica).

The proximal caecum in quails consists of lymphoid and non-lymphoid structures. The caecal tonsils in the proximal part of the caecum are units of gut-associated lymphoid tissue in poultry. This study aimed to examine the histological characteristics of the proximal caecum, as well as compositions of dendritic cells (DCs) and antigen-presenting cells (APCs) in the caecal tonsil of quails. Tissue sections were stained with Crossman's triple, periodic acid-Schiff, Gordon and Sweet's silver, Congo red and methyl green-pyronin dyes, as well as immunohistochemically by the streptavidin-biotin-peroxidase complex method. Caecal lymphoid tissue was located in the lamina propria and submucosa. Germinative centres were observed within the lymphoid tissue. Reticular fibres were mainly distributed in the border area of the germinal centre with only a few fibres scattered in the centre. Plasma cells were observed in the subepithelial region and germinal centres. Eosinophil granulocytes were prevalent in the lymphoid tissue. Additionally, CD83-immunoreactive DCs and MHC class II immunoreactive APCs were present in the subepithelial area and diffuse lymphoid tissue. While DCs were seen in the germinal centres of tonsillar units, APCs were rarely present in the germinal centres, but they were noticed around the germinal centres. In conclusion, the histological structure of the proximal caecum in quails and the distributions of some immunological cells in the caecal tonsils were revealed. Therefore, the defensive role of the caecal tonsils in the digestive system may be better understood, and comparative studies may be carried out.

Human adenovirus replication and persistence in hypertrophic adenoids and palatine tonsils in children.

The role of human adenovirus (HAdV) infection in different acute diseases, such as febrile exudative tonsillitis, conjunctivitis, and pharyngoconjunctival fever is well established. However, the relationships, if any, of HAdV persistence and reactivation in the development of the chronic adenotonsillar disease is not fully understood. The present paper reports a 3‐year cross‐sectional hospital‐based study aimed at detecting and quantifying HAdV DNA and mRNA of the HAdV hexon gene in adenoid and palatine tonsil tissues and nasopharyngeal secretions (NPS) from patients with adenotonsillar hypertrophy or recurrent adenotonsillitis. HAdV C, B, and E were detectable in nearly 50% of the patients, with no association with the severity of airway obstruction, nor with the presence of recurrent tonsillitis, sleep apnea or otitis media with effusion (OME). Despite the higher rates of respiratory viral coinfections in patients with HAdV, the presence of other viruses, including DNA and RNA viruses, had no association with HAdV replication or shedding in secretions. Higher HAdV loads in adenoids showed a significant positive correlation with the presence of sleep apnea and the absence of OME. Although this study indicates that a significant proportion (~85%) of individuals with chronic adenotonsillar diseases have persistent nonproductive HAdV infection, including those by HAdV C, B, and E, epithelial and subepithelial cells in tonsils seem to be critical for HAdV C production and shedding in NPS in some patients, since viral antigen was detected in these regions by immunohistochemistry in four patients, all of which were also positive for HAdV mRNA detection.

Streptococcal superantigen-induced expansion of human tonsil T cells leads to altered T follicular helper cell phenotype, B cell death and reduced immunoglobulin release.

SummaryStreptococcal pyrogenic exotoxin (Spe) A expression is epidemiologically linked to streptococcal tonsillo‐pharyngitis and outbreaks of scarlet fever, although the mechanisms by which superantigens confer advantage to Streptococcus pyogenes are unclear. S. pyogenes is an exclusively human pathogen. As the leucocyte profile of tonsil is unique, the impact of SpeA production on human tonsil cell function was investigated. Human tonsil cells from routine tonsillectomy were co‐incubated with purified streptococcal superantigens or culture supernatants from isogenic streptococcal isolates, differing only in superantigen production. Tonsil cell proliferation was quantified by tritiated thymidine incorporation, and cell surface characteristics assessed by flow cytometry. Soluble mediators including immunoglobulin were measured using enzyme‐linked immunosorbent assay. Tonsil T cells proliferated in response to SpeA and demonstrated typical release of proinflammatory cytokines. When cultured in the absence of superantigen, tonsil preparations released large quantities of immunoglobulin over 7 days. In contrast, marked B cell apoptosis and abrogation of total immunoglobulin (Ig)A, IgM, and IgG production occurred in the presence of SpeA and other superantigens. In SpeA‐stimulated cultures, T follicular helper (Tfh) cells showed a reduction in C‐X‐C chemokine receptor (CXCR)5 (CD185) expression, but up‐regulation of OX40 (CD134) and inducible T cell co‐stimulator (ICOS) (CD278) expression. The phenotypical change in the Tfh population was associated with impaired chemotactic response to CXCL13. SpeA and other superantigens cause dysregulated tonsil immune function, driving T cells from Tfh to a proliferating phenotype, with resultant loss of B cells and immunoglobulin production, providing superantigen‐producing bacteria with a probable survival advantage.

Diverging incidence trends for larynx and tonsil cancer in low socioeconomic regions of the US

ObjectiveOropharynx cancer incidence trends in low socioeconomic (SES) regions of the United States (US) have not been well described. Our objective was to describe tonsil cancer incidence trends in low SES regions, and compare observed trends with those for larynx cancer. Materials and methodsAge-adjusted incidence rates and trends for tonsil and larynx squamous cell carcinomas (2000–14) from Surveillance, Epidemiology, and End Results (SEER 18) were evaluated using SEER*Stat and Joinpoint 4.5.0.1. Annual percentage changes (APCs) were compared between low and high SES counties. The laryngeal cancer cohort was included as a comparator reflecting a tobacco-related malignancy. ResultsTonsil cancer incidence trends increased at least as much in low SES as in high SES counties (APC/AAPC 4.4, 95%CI 2.4–6.4 versus APC/AAPC 2.9, 95%CI 2.4–3.3). Pairwise comparison confirmed no differences between incidence trends across SES quintiles for tonsil cancer incidence rates. In contrast, age-adjusted incidence rates of larynx cancer decreased in high SES counties (APC/AAPC −2.4, 95%CI −2.4 to −2.0, p < 0.001) and were stable in low SES counties (APC/AAPC −0.9, 95%CI −1.9 to 0.2, p = 0.10). Compared with larynx cancer patients, tonsil cancer patients in low SES regions were significantly more likely to be younger and white. ConclusionIn low SES US counties, tonsil cancer incidence rates increased from 2000 to 2014, while larynx cancer rates did not change, reflecting diverging trends for larynx and tonsil cancers. Tonsil cancer incidence rates are increasing in most US regions regardless of regional socioeconomic status. Prevention efforts should take these findings into account.

Revisiting the Tissue Microenvironment of Infectious Mononucleosis: Identification of EBV Infection in T Cells and Deep Characterization of Immune Profiles.

To aid understanding of primary EBV infection, we have performed an in depth analysis of EBV-infected cells and of local immune cells in tonsils from infectious mononucleosis (IM) patients. We show that EBV is present in approximately 50% of B-cells showing heterogeneous patterns of latent viral gene expression probably reflecting different stages of infection. While the vast majority of EBV+ cells are B-cells, around 9% express T-cell antigens, with a predominance of CD8+ over CD4+ cells. PD-L1 was expressed by a median of 14% of EBV+ cells. The numbers of EBER+PD-L1+ cells were directly correlated with the numbers of EBER+CD3+ and EBER+CD8+ cells suggesting a possible role for PD-L1 in EBV infection of T-cells. The microenvironment of IM tonsils was characterized by a predominance of M1-polarized macrophages over M2-polarized cells. However, at the T-cell level, a heterogeneous picture emerged with numerous Th1/cytotoxic cells accompanied and sometimes outnumbered by Th2/regulatory T-cells. Further, we observed a direct correlation between the numbers of Th2-like cells and EBV– B-cells. Also, a prevalence of cytotoxic T-cells over Th2-like cells was associated with an increased viral load. These observations point to contribution of B- and Th2-like cells to the control of primary EBV infection. 35% of CD8+ cells were differentiated CD8+TBET+ cells, frequently detected in post-capillary venules. An inverse correlation was observed between the numbers of CD8+TBET+ cells and viral load suggesting a pivotal role for these cells in the control of primary EBV infection. Our results provide the basis for a better understanding of immune reactions in EBV-associated tumors.

Tonsil conventional dendritic cells are not infected by porcine reproductive and respiratory syndrome virus

Porcine reproductive and respiratory syndrome virus (PRRSV) infects monocyte-derived DCs, and previous reports have shown that PRRSV does not infect conventional DCs (cDCs) in vitro, but the effects on cDCs from lymphoid tissues are unknown. This study analyzed the response and susceptibility of tonsil DEC205+cDCs from infected pigs. We confirmed the phenotype and lineage of bona fide tonsil cDCs with the mRNA expression of FLT3+ and the phenotype MHCII+CADM1highDEC205+ (DEC205+cDCs). These cells were not infected by PRRSV, whereas CD163+ tonsil cells were infected. The numbers of tonsil cDCs and CD163+ cells were not affected by PRRSV, in contrast to the reduction in alveolar macrophage numbers. DEC205+cDCs exhibited an increase in the expression of IL-12 at 5 days postinfection, suggesting a proinflammatory response by these cells to the virus. In summary, this study confirms that, in vitro and in vivo, cDCs are not susceptible to PRRSV but can respond against it.

Purification, characterization and in vitro and in vivo immune enhancement of polysaccharides from mulberry leaves.

Mulberry leaf polysaccharide (MLP) was extracted and purified by DEAE-52 cellulose and Sephadex G-100 column chromatography to afford two major purified polysaccharides (MLP-1 and MLP-2). The purified polysaccharides were characterized, and their immune-enhancing properties were investigated. MLP-1 had a molecular weight of 9.31×104 Da and was composed of mannose, rhamnose, glucose, galactose, xylose, and arabinose in a molar ratio of 0.71:1.00:2.76:1.13:3.70:2.81. The molecular weight of MLP-2 was 2.22×106 Da, and its monosaccharide constituents were mannose, rhamnose, glucose, galactose, and arabinose in a molar ratio of 1.31:8.45:6.94:1.00:11.96. Infrared spectroscopy showed that each MLP had a typical absorption peak characteristic of sugars, and ultraviolet (UV) spectroscopy showed that neither MLP contained nucleic acid or protein components. Then, the abilities of these polysaccharides to stimulate spleen lymphocyte proliferation in mice in vitro were compared by the 3-[4,5-dimethylthiazol-2-yl]-2,5-diphenyltetrazolium bromide (MTT) assay. MLP-2 was more effective than MLP-1; therefore, MLP-2 was chosen for the study of its immune-enhancing effects in vivo. For the in vivo experiments, 14-day-old chickens immunized with Newcastle disease (ND) vaccine were orally administered MLP-2, and Astragalus polysaccharide (APS) was used as the control. Each chicken was orally administered 4 mg or 8 mg of MLP-2 for seven consecutive days starting three days before ND vaccine immunization. MLP-2 significantly improved the ND serum antibody titer and interleukin-2 (IL-2), interferon-γ (IFN-γ) and immunoglobulin A (sIgA) concentrations in tracheal and jejunal wash fluids, and increasing numbers of immune globulin A-positive (IgA+) cells in cecal tonsils and increased body weight. These results indicated that MLP-2 could significantly enhance immune activity and could therefore be utilized as an immunopotentiator drug candidate.

Construction and preliminary identification of a prokaryotic expression single-chain antibody fragments library against Streptococcus pneumoniae from antibody-producing cells in human tonsil

ABSTRACT Tonsillitis is the inflammation of the tonsils due to infection, many patients ultimately have to undergo tonsillectomy. In order to improve the accuracy of diagnosis and even create a new treatment for tonsillitis, we constructed a prokaryotic expression single-chain antibody fragment library against Streptococcus pneumoniae with immunoglobulin heavy chain variable region (VH), κ light chain (Vκ), and λ light chain (Vλ) genes by using human tonsil tissue. Plasmid DNA sequencing showed that single-chain antibodies were complete and constructed correctly. The binding activity of recombinant clones was detected by enzyme-linked immunosorbent assay (ELISA), results showed that the binding activity and specificity of anti-S. pneumoniae single-chain fragment variable (scfv) is proved to be successful. The single-chain antibody may be an attractive strategy for tonsillitis etiologic diagnosis and therapy.

Investigations of relationships between alterations of the gastrointestinal tract caused by feeding variously processed feedstuffs and blood and immunological traits of broilers

The influence of variously processed feedstuffs on the gastrointestinal tract, various blood parameters, and T cell subsets of different localizations was investigated in 36 broilers. Birds were fed a similarly composed feed mixture differing in particle size (coarse vs. fine). Feedstuffs with different particle size distribution were additionally hydrothermally treated (HTT) (without-non-compacted, pelleted, expanded, and pelleted). The HTT affected BW at slaughtering day and feed intake (P < 0.001). Birds fed non-compacted feed consumed less feed and were lighter than animals of other feeding groups irrespective of grinding fineness. Proventricular size was influenced by HTT (P < 0.001) but not the weight. Generally, feeding of pelleted feed increased proventricular size. Gizzard weight and length was affected by particle size and HTT (P < 0.01). Non-compacted feed resulted in heavier but shorter gizzards compared to other feeds except for finely ground expanded and pelleted feed. Organ traits of gizzard and proventriculus were strongly related to the feed intake. The jejunum was heavier in birds fed expanded and pelleted feed compared to animals fed solely pelleted feed (P = 0.030). The intestinal segment weight to length ratio (WL ratio) of duodenum, jejunum, and total small intestine was affected by HTT (P < 0.01). Generally, non-compacted feed resulted in lowest WL ratios regardless of the particle size distribution. Feeding variously processed feedstuffs also affected white blood cells, blood sodium, and chloride concentrations (P < 0.05) and tended to influence blood carbon dioxide partial pressure and blood total carbon dioxide levels (P < 0.1) in venous blood. Subsets of CD4/CD8 double negative T cells of caecal tonsils were influenced by HTT (P < 0.043). Non-compacted feed tended to increase these subsets in this localization compared to other feeds. In conclusion, the present study shows that an increased feed intake provoked by feed compaction evoke proventricular and ventricular alterations. Furthermore, feeding of variously processed feedstuffs influences the acid base homeostasis and the local immune system of caecal tonsils.

Effects of pepsin and pepstatin on reflux tonsil hypertrophy in vitro.

There is evidence that pepsin can aggravate tonsil hypertrophy. Pepstatin is a potent inhibitor of pepsin activity and could protect patients against reflux tonsil hypertrophy by inhibiting pepsin. We examined the effects of pepstatin on the development of tonsil hypertrophy to investigate pepsin’s role in the pathogenesis of tonsil lesions. We investigated whether pepstatin suppresses pepsin-mediated lymphocyte proliferation in tonsil hypertrophy. Forty-nine children with tonsil hypertrophy and twenty-two adults with tonsillitis were recruited to the study prior to surgery. Tonsil tissue from each patient was harvested and assessed for changes in the number of lymphocytes and macrophages in the presence of pepsin and pepstatin. We found that the proportions of CD4- and CD14-positive cells were significantly lower (p < 0.05), but that the proportions of CD19- and CD68-positive cells were significantly higher (p < 0.05), in children than in adults. There were significantly more CD4-positive cells after pepsin treatment, but these numbers were reduced by pepstatin. The levels of both interleukin-2 (IL-2) and interferon gamma (IFN-γ) increased significantly in response to pepsin, but were reduced when pepsin was inhibited by pepstatin. The level of IL-10 is reduced in pepsin-treated CD4 cells and the level is restored by pepstatin. IL-2 blocking reduced the increased CD4 cell number by pepsin. But, an additive or a synergic effect is not founded in combined with IL-2 blocking and pepstatin. Pepsin-positive cells did not co-localize with CD20 and CD45 cells, but they were found surrounding CD20- and CD45-positive hypertrophic tonsil cells. Pepsin-positive cells co-localized with CD68-positive cells. It is probable that pepsin from extraesophageal reflux aggravates tonsil hypertrophy and pepstatin exerts a protective effect by inhibiting pepsin activity.

CD40 signaling—germinal center b cells go at their own pace

CD40 signaling—germinal center b cells go at their own pace

Low-Level Laser Therapy Stimulates Proliferation in Head and Neck Squamous Cell Carcinoma Cells.

Objectives: Low-level laser therapy (LLLT) is a promising non-invasive treatment option for oropharyngeal mucositis, which is a common side effect of many oncological treatments. LLLT is known for its wound healing properties due to the stimulation of cellular processes, such as proliferation, migration and differentiation. Controversy exists on the possible stimulatory effect of LLLT on head and neck cancer (HNSCC) cells in patients treated with radiotherapy. The aim of this study was to evaluate the biostimulatory effect together with the underlying mechanisms of LLLT on HNSCC cancer cells and normal epithelial cells.Materials and methods: HNSCC cell lines (SCC154, SQD9, and SCC61) and human tonsil epithelial cells were exposed to a Gallium-Aluminum-Arsenide diode laser (830 nm, 150 mW) with energy densities of 0, 1, and 2 J/cm2. The proliferation potential of the cells was assessed by Sulforhodamine B assay, immunoblotting (mitogenic pathways), immunocytochemistry (Ki67), and flow cytometry (PI cell cycle staining).Results: Cell proliferation was increased in HNSCC cell lines after laser irradiation with 1 J/cm2, whereas no significant increase was seen after laser irradiation with 2 J/cm2. In contrast, no effect on cell proliferation was seen in the human tonsil epithelial cells after laser irradiation with any of the energy densities. The increased proliferation was associated with elevated levels of pAKT, pERK, and Ki67 protein expression and cell cycle progression.Conclusion: Our results show that LLLT increases cell proliferation in a dose-dependent manner in HNSCC cells but not in normal epithelial tonsil cells. These results suggest that LLLT has to be used with caution when treating oropharyngeal mucositis in HNSCC patients since tumor cells present in the LLLT irradiation field could be triggered by LLLT.

The interplay between local immune response and Epstein-Barr virus-infected tonsillar cells could lead to viral infection control.

Epstein Barr virus (EBV) gains access to the host through tonsillar crypts. Our aim was to characterize microenvironment composition around EBV+ cells in tonsils from pediatric carriers, to disclose its role on viral pathogenesis. LMP1 expression, assessed by immunohistochemistry (IHC), was used to discriminate EBV + and - zones in 41 tonsil biopsies. Three regions were defined: Subepithelial (SE), interfollicular (IF) and germinal center (GC). CD8, GrB, CD68, IL10, Foxp3, PD1, CD56 and CD4 markers were evaluated by IHC; positive cells/100 total cells were counted. CD8+, GrB+, CD68+ and IL10+ cells were prevalent in EBV+ zones at the SE region (p < 0.0001, p = 0.03, p = 0.002 and p = 0.002 respectively, Wilcoxon test). CD4+ and CD68+ cell count were higher in EBV + GC (p = 0.01 and p = 0.0002 respectively, Wilcoxon test). Increment of CD8, GrB and CD68 at the SE region could indicate a specific response that may be due to local homing at viral entry, which could be counterbalanced by IL10, an immunosuppressive cytokine. Additionally, it could be hypothesized that CD4 augment at the GC may be involved in the EBV-induced B-cell growth control at this region, in which macrophages could also participate.