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Immunohistochemical identification of T- and B-lymphocytes delineated by the unlabelled antibody enzyme method. III. Topographical and quantitative distribution of T- and B-cells in human palatine tonsils.

T-lymphocytes and B-lymphocytes are identified in tissue sections of human tonsils by applying the unlabelled antibody enzyme method. The epithelium of the tonsils contains a majority of immunoglobulin-positive cells and fewer T-lymphocytes. In the subepithelial zones, areas composed of B-cells predominate, however, regions containing T-lymphocytes are also present. The latter are mainly arranged in the lamina propria around high-endothelial venules and often include plasma cells containing immunoglobulin. Follicles containing germinal centres display a complex structure which changes during development. The lymphocytic cap consists of densely packed lymphocytes, labelled heavily by anti-IgM and anti-IGD, and of individual T-lymphocytes. Germinal centres show a framework of immunoglobulin-positive dendritic reticular cells; they contain some heavily labelled lymphoid cells and several cells weakly labelled by anti-IgM and anti-IgA, as well as a small number of T-lymphocytes. Furthermore, the total areas of T- and B-lymphocytes measured by planimetry may differ considerably between different tonsils. Especially total areas of germinal centres vary to a great extent. The quantitative data on amounts of T- and B-cells achieved by planimetry are comparable to those reported in cellular suspensions of tonsils.

B-Lymphocytes↗

The emigration of lymphocytes from palatine tonsils after local labelling.

Lymphocytes in the palatine tonsils of normal young pigs were selectively labelled by minute multiple injections of fluorescein isothiocyanate into the tonsils. One day later the numbers of tonsil-derived lymphocytes were determined in cervical, bronchial and mesenteric lymph nodes, spleen, Peyer's patches, thymus, bone marrow and blood. Although not all tonsillar lymphocytes could be labelled by this technique, a total of approximately 0.6 X 10(9) lymphocytes emigrated from the tonsils. Relatively more lymphocytes were found in lymph nodes than in the spleen and very few in the thymus, Peyer's patches and bone marrow. This organ distribution was different from the results from selectively labelling lymphocytes in lymph nodes, spleen and bone marrow.

Animals↗

Identification of M-cells in the rabbit tonsil by vimentin immunohistochemistry and in vivo protein transport.

The rabbit palatine tonsil was studied by electron microscopy to determine whether M-cells similar to those of the Peyer's patches exist in the tonsil epithelium. A subpopulation of crypt epithelial cells was found with long, irregularly shaped microvilli, small cytoplasmic vesicles and engulfing intraepithelial lymphocytes and macrophages. Using ultrastructural immunohistochemistry, vimentin, the rabbit maker for intestinal and bronchial M-cells, was detected in the cytoplasm of these cells, whereas no vimentin immunoreactivity was found in the remaining epithelial cells. The vimentin filaments surrounded the nucleus of the presumed M-cells and lay beneath the plasma membrane that surrounded intraepithelial lymphocytes. In vivo experiments using horseradish peroxidase as tracer revealed that this protein was endocytosed by the presumed M-cells and transported to the intercellular spaces between epithelial cells and lymphocytes. The results indicate that specialized epithelial cells exist in the tonsil which have morphological characteristics similar to those of intestinal M-cells, are in close contact to cells of the immune system, are positive for the rabbit M-cell marker vimentin, and are capable of antigen uptake and transcytosis. It is therefore concluded that M-cells are present in the tonsil and probably play a role in the initiation of immune responses to orally delivered antigens.

Animals↗

Involvement of Epstein-Barr virus (EBV)-encoded latent membrane protein (LMP)-1 in the potentiation of C epsilon mRNA expression in human tonsil-derived cells.

The aim of our study was to investigate a possible correlation between Epstein-Barr virus (EBV) infection status, including its latent gene expression, and expression of allergy-related genes in human tonsil-derived cells. In the tonsil-derived cells from the patients undergoing routine tonsillectomies for palatine tonsil hypertrophy or tonsillar focal infection, the presence of EBV DNA and mRNA expressions of latent membrane protein (LMP)-1, C epsilon chain, and activation-induced cytidine deaminase (AID) were detected by PCR and RT-PCR analysis, respectively. Of all the 12 patients, PCR products amplified from EBV DNA BamHI W fragment were detected in the tonsils from the 10 patients (83.3%). LMP1 mRNA expressions were confirmed in the six patients (50%). Both LMP1 mRNA expressions and EBV DNA were detected in the five patients. EBV DNA, but not LMP1 mRNA expression, was detected in the five patients. LMP1 mRNA expression, but not EBV DNA, was detected in one patient. In one patient, neither EBV DNA nor LMP1 mRNA expression was confirmed. C epsilon mRNA expressions were confirmed in all the 12 patients along with AID mRNA expressions. The degree of C epsilon mRNA expression, however, varied with the patients. The Fisher's exact probability test revealed a statistically significant correlation between LMP1 and C epsilon gene expressions, indicating that C epsilon mRNA expression level was significantly higher in the LMP1 positive samples than in the negative samples. On the other hand, there was no significant correlation between AID and LMP1 mRNA expressions. Thus, EBV infection is a notable factor capable of exacerbating allergic inflammation.

Adolescent↗

Intercellular and lymphatic pathways associated with tonsils of the soft palate in young pigs.

Tonsils of the soft palate of pigs are the main oropharyngeal lymphoid tissues that protect the body against antigens entering through the mouth. The aim of this work was to elucidate the intercellular and lymphatic pathways by which lymph and cells are transported through these tonsils. Tonsillar tissue from freshly-killed pigs was examined using light microscopy and electron microscopy, or was injected with Mercox for scanning electron microscopy of corrosion casts. Intercellular fluid passes between epithelial cells and is continuous with that of the subepithelium. Fluid from the subepithelium flows into sinuses that form a network around the apex of follicles. These sinuses are continuous with parafollicular sinuses that penetrate the parafollicular tissue between the follicles. Some parafollicular sinuses are traversed by a complex network of cell processes, whereas others appear to lack such processes. Some parafollicular sinuses are closely located (10 microm) to venules; others lie adjacent to the follicle capsule. No lymphatics enter or leave the follicles. All lymph from the tonsils must traverse parafollicular sinuses before entering septal vessels, and these are continuous with basal vessels. Basal vessels coalesce to form efferent vessels that transport lymph from the tonsil to the primary lymph nodes. Septal, basal and efferent lymphatic vessels contain prominent valves and many lymphocytes. Lymphatic sinuses appear to be a significant pathway for lymphocytes migrating from the tonsillar lymphoid tissue.

Animals↗

Induction of IgM production by pokeweed mitogen and interleukin-2: different responses by human peripheral blood cells and tonsil cells.

Stimulation of tonsils or peripheral blood lymphocytes with either pokeweed mitogen (PWM) or interleukin-2 (IL-2) results in significant IgM production. However, the combination of PWM and IL-2 shows a synergistic IgM synthesis in tonsil cells, whereas the secretion in peripheral blood cells is diminished compared to activation with either stimulus alone. The heavy-chain isotype distribution does not significantly change in the two cell types when stimulated with either PWM, IL-2, or the combination. The Ig synthesis by tonsil cells to PWM or IL-2 drastically increases in the presence of monocytes, but the response to PWM + IL-2 together is not affected. The reason for the synergistic IgM production to PWM + IL-2 in tonsil cells is the relative lack of monocytes and the low number of T8+-suppressor cells. The decreased IgM production in peripheral blood cells after stimulation with both PWM and IL-2 is due to the presence of T8+ cells. These data show that the monocyte-dependent IL-2 production is an essential step in PWM-induced Ig secretion; the subsequent induction of the T-cell-helper signal for B-cell differentiation is determined by the balance between the strength of the stimulus for T cells and the T4/T8 ratio. Hence, these parameters should be included when the B-cell function is studied in vitro with the use of PWM and IL-2.

Antigen-Presenting Cells↗

Lymphocyte subsets in human adenoids and tonsils. Rosette formation, alpha-naphthyl acetate esterase cytochemistry, monoclonal antibodies and peanut lectin reactivity.

The distribution of mononuclear cell subsets has been studied in human adenoids, tonsils and peripheral blood (PB) by evaluating the presence of surface immunoglobulins, E-rosette formation, receptors for IgG Fc and for complement, alpha-naphthyl acetate esterase (ANAE) cytochemistry, reactivity with peanut lectin (PNA) and with monoclonal antibodies (McAb) (OK panel). Adenoids and tonsils, compared to PB, contain (1) fewer macrophages and T cells but more B cells; (2) higher proportions of ANAE negative, complement receptors and Ia-like antigens bearing T lymphocytes; (3) higher percentages of cells reacting with the McAbs OKT9 and OKT10 ("immature" lymphoid cells). In both adenoids and tonsils, clusters, formed by a central heavily ANAE stained interdigitating cell surrounded by lymphocytes with a sickle-shaped ANAE reaction, were found. Analogous clusters have been previously described in mice and human thymus. Two major hypotheses could be put forward: (1) adenoids and tonsils contain "immature" lymphoid cells undergoing education process, or (2) the above organs contain lymphocytes activated by a constant exposure to bacterial antigens or mitogens.

Adenoids↗

Antibody-producing cells in peripheral blood and tonsils after oral treatment of children with bacterial ribosomes.

The efficacy of ribosomal preparations as mucosal immunostimulants was examined in the peripheral blood and tonsils of 14 children, before and after 28 days of oral treatment with D-53, a preparation of ribosomes from Klebsiella pneumoniae, Streptococcus pneumoniae, Haemophilus influenzae and Streptococcus pyogenes. Tonsils from 10 untreated children were used as controls. Immunofluorescence and ELISAspot were performed to analyse variations in the numbers of immunoglobulin-containing and immunoglobulin-secreting B-cells. Both isotypic and antigenic specificities of these two types of cells were investigated. Significant differences were observed after treatment in the peripheral blood as well as between tonsils from treated and untreated children. In the peripheral blood a significant increase in immunoglobulin-secreting cells directed against antigenic specificities of D-53 was the major change. In tonsils, higher numbers of specific immunoglobulin-containing and secreting cells, and higher numbers of IgA-secreting cells were induced in treated children. These data support the efficacy of D-53 as an oral immunostimulant.

Administration, Oral↗

Pharyngeal and tonsil infections caused by non-group A Streptococcus.

Non-group A streptococci are members of the genus Streptococcus but do not share the notoriety of their cousin from group A. Most physicians, including otolaryngologists and head and neck surgeons, do not associate them with upper respiratory tract and head and neck infections. Some laboratories do not bother to report their presence on culture. At the University of California, Los Angeles they have been shown to cause (1) tonsillitis, painful tonsils lacking exudate and erythema (group C, one case; group F, one case), (2) acute nonexudative tonsillitis (group B, two cases; group C, one case; group F, one case), and (3) acute exudative tonsillitis (group C, one case). It should be remembered when there is a report of their presence that they are usually vulnerable to penicillin and its analogs. By judicious use of these drugs, morbidity can be diminished.

Adult↗

Inhibition of virus replication in the tonsils of pigs previously vaccinated with a Chinese strain vaccine and challenged oronasally with a virulent strain of classical swine fever virus.

After an oronasal (O.N.) infection with classical swine fever (C.S.F.) virus, virus multiplication can be detected in the tonsils from Day 2 post infection (p.i.) till death. The course of viral replication during the first 10 days after O.N. challenge exposure of pigs, previously vaccinated with a Chinese strain vaccine in the presence or absence of maternal antibodies, was studied using direct immunofluorescence techniques on cryostat sections and virus isolations. When piglets were challenged O.N. in the presence of maternal antibodies, virus replication in the tonsils still occurred. The multiplication period and the localization of the virus, however, were directly correlated to the maternal antibody levels. The maternal antibody level also seems responsible for the efficacy of the vaccination to prevent challenge virus replication in the tonsils: vaccination in the presence of low maternal antibody titers completely inhibited virus replication; vaccination in the presence of high maternal antibody titers only reduced the multiplication period of the O.N.-administered virulent virus. In both cases, animals were challenged 1 week post vaccination. Vaccination of seronegative animals resulted in an almost complete inhibition of the virus replication in the tonsils during a full fattening period: cryostat sections revealed a limited virus replication in three out of 20 animals. In one of these animals, virus replication was probably so negligible that virus isolation remained negative.

Animals↗

Increase of activated T-cells and up-regulation of Smad7 without elevation of TGF-beta expression in tonsils from patients with pustulosis palmaris et plantaris.

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.

Adult↗

Development of the palatine tonsil in conventional and germ-free piglets.

Palatine tonsils, like the Peyer's patches, are considered to be major inductive sites for the mucosa-associated lymphoid tissue (MALT), providing sampling and effector functions for the upper respiratory tract. Consistent with this, they have the architecture required of a classic inductive site (B-cell follicles, immunoglobulin class switching and the presence of naïve and memory T-cells). Here we show that much of this architecture develops after birth in the neonatal piglet, the numbers of T-cells, B-cells and accessory cells increasing with age. Conventional piglets also had higher levels of activated and memory T-cell subsets than germ-free piglets, consistent with development occurring as a result of microbial stimulus. The results suggest that the microbial environment influences the development of the tonsil immunological architecture. Given the role of the tonsil in induction of mucosal responses, this raises questions as to the effectiveness of the tonsil in dealing with colonising organisms in the neonate.

Age Factors↗

Search for Herpesvirus 1 and 2 by in situ hybridization in tonsils and adenoids.

OBJECTIVE: Herpes simplex virus (HSV) has been described as cause of acute tonsillitis. It has also been found in nasopharyngeal florid lymphoid infiltrate, mostly composed of CD4+, CD56+ T-cells, simulating lymphoma. In spite of its widespread prevalence in latent form, to the best of our knowledge no study is available on in situ detection of HSV in chronically hyperplastic nasopharyngeal lymphoid tissue. The purpose of the present study was to search for the presence of HSV 1 and 2 in 21 adenoids and 15 tonsils from children (2-12 years of age) in which these organs had been surgically removed due to hypertrophy. METHODS: Paraffin wax-embedded sections from the 36 cases were submitted to the in situ hybridization technique, using the biotinilated probe to Herpes simplex virus 1 and 2 (Pan Path, Amsterdam) and the Rembrandt Universal DISH & HRP Detection Kit (Pan Path, Amsterdam). Positive control consisted of a previously tested Herpes infected lung. RESULTS: In none of the 36 cases studied were positive nuclei detected in adenoid and tonsils, either in lymphoid, in stroma or in epithelial cells, as those seen in the positive control. CONCLUSION: HSV does not seem to be implied in tonsil or adenoid chronic lymphoid hyperplasia. These organs do not seem to harbor the virus latently, or the amount of virus is too low to be detected without amplification methods.

Adenoids↗

Cytogenetic biomonitoring in children with chronic tonsillitis: micronucleus frequency in exfoliated buccal epithelium cells.

OBJECTIVE: To investigate the possible harmful cytogenetic effects associated with chronic tonsillitis by analyzing the micronucleus frequency and other nuclear abnormalities in exfoliated buccal epithelial cells. MATERIALS AND METHODS: The study consisted of 20 children with chronic tonsillitis, and 20 control subjects with similar age and sex. The ages ranged between 5 and 12 years old (mean age: 7.5). The patients were diagnosed as having chronic tonsillitis on the basis of history, throat culture and clinical examinations. Buccal cell samples were collected with a wooden spatula. The samples were then applied to clean microscope slides. Smears were air dried and fixed in methanol:acetic acid. Then slides were stained by the Feulgen reaction technique. Three slides were prepared for each subject and 1000 cells were evaluated per slide to determine the frequencies of micronucleus and other nuclear abnormalities (binucleats, karyorrhexis and karyolysis). Statistically, Mann-Whitney U-test was used to analyze and compare the data. RESULTS: The mean micronucleus frequencies in patient and control groups were 5.29+/-1.67 and 1.58+/-0.33, respectively. In the patient group, mean binucleus, karyorrhexis and karyolysis frequencies were 3.13+/-1.2, 2.04+/-0.64, and 1.74+/-0.47, respectively. However, in the control group, mean binucleus, karyorrhexis and karyolysis frequencies were 1.43+/-0.47, 1.26+/-0.45, and 0.88+/-0.27, respectively. The mean frequencies of all parameters in the patient group were higher than the control values, and the difference was found to be statistically significant (p<0.001). CONCLUSION: Our results revealed that children with chronic tonsillitis could be under risk of significant cytogenetic damage.

Case-Control Studies↗

Pre-surgical child behavior ratings and pain management after two different techniques of tonsil surgery.

OBJECTIVE: The purpose of this investigation was to compare child behavior before surgery with experience of pain and anxiety in relation to two techniques of tonsil surgery, to relate previous experiences of surgery/tonsillitis with anxiety and pain, and to compare the children's, parent's and nurse's rating of pain. METHOD: Ninety-two children (5-15 years) with sleep-disordered breathing (SDB) and with or without recurrent tonsillitis were randomized to partial tonsil resection/tonsillotomy (TT) or full tonsillectomy (TE). MEASURES: Parents: Child Behavior Checklist (CBCL). Children: State-Trait-Anxiety Inventory for Children (STAIC) and seven-point Faces Pain Scale (FPS). Parents/staff: seven-point Verbal Pain Rating Scale (VPRS). Pain relievers were opoids, paracetamol and diclophenac. RESULTS: These children with SDB scored significantly higher on CBCL than did normative groups, but no connection was observed between CBCL rating and experience of pain. There was no relation between pre-operative anxiety and pain. The post-operative anxiety level (STAIC) correlated with pain. The TE-group scored higher on STAIC after surgery. Previous experience of surgery or tonsillitis did not influence post-operative pain. The TE-group rated higher experience of pain despite more medication. The nurses scored pain lower than the parents/children and under-medicated. CONCLUSION: SDB may influence children's behavior, but with no relation to post-operative pain. The surgical method predicts pain better than does the child's behavior rating. The nurses underestimated the pain experienced by the child.

Acetaminophen↗

Use of a novel serum ELISA method and the tonsil-carrier state for evaluation of Mycoplasma hyosynoviae distributions in pig herds with or without clinical arthritis.

A novel indirect ELISA test using deoxycholate extracted antigens coated onto a hydrophobic polystyrene surface for measurement of serum antibodies specific for Mycoplasma hyosynoviae was developed. Sensitivity and specificity of the test were found to be superior to previous ELISAs as tested on porcine serum following experimental Mycoplasma infections as well as by analysis of samples from one Danish herd known to be free of M. hyosynoviae and samples from two Norwegian herds without clinical suspicion of M. hyosynoviae infections since their establishment. The epidemiology of M. hyosynoviae infection was then investigated in Danish pig herds with evidence of clinical M. hyosynoviae arthritis (MhA herds, n = 4) and in herds with M. hyosynoviae-carriers among slaughter pigs, but with limited clinical lameness (MhC herds, n = 4). M. hyosynoviae bacteriaemia and tonsil-carrier state were determined by culture of cross-sectional samples of whole-blood (n = 238) and tonsil scrapings (n = 322), respectively. Levels of serum antibodies (n = 396) were measured by the novel indirect ELISA test. There was no significant difference in the ELISA results between the MhA and the MhC herds. Pigs that were tonsil-carriers had a significantly higher response in the ELISA test (P < 0.001) than non-carriers. Slaughter pigs showed higher ELISA values (P < 0.001) and they were more prone to be tonsil-carriers (P < 0.001). The most critical period for spread of the infection seems to be the nursery period (4-12 weeks). The results indicate that M. hyosynoviae infection progresses similarly in herds irrespective of the presence of clinical arthritis. Thus, clinical arthritis due to M. hyosynoviae is probably triggered by other host or herd factors than low levels of serum antibodies or by differences in virulence factors between M. hyosynoviae strains.

Animals↗

[Tonsillar mast cell infiltration and allergic tonsillitis].

We report the case of a seven year-old boy bearing of exudative acute tonsillitis after strawberry ingestion with no previous infectious history. After having diagnosed it as hypersensitivity type I, this patient began to suffer repetitive episodes of exudative and purulent tonsillitis, related to cool drinks and direct manipulation of tonsils. These were hyperplastic and Staphylococcus sp. and Streptococcus sp. were isolated in them. The episodes were shortened by the use of corticosteroids. Tonsillectomy was the definitive therapy. Biopsy informed of mast cells infiltration in germinal centers of both palatine tonsils.

Biopsy↗

Rapid diagnosis of adenoviral tonsillitis: a prospective clinical study.

Adenovirus antigen was detected by radioimmunoassay in the nasopharyngeal specimens of 30 of 74 children with febrile exudative tonsillitis. In the other 44 children, the cause of tonsillitis was identified in 16 (beta-hemolytic streptococci or Epstein-Barr, parainfluenza, or herpes simplex viruses). The clinical picture of adenoviral tonsillitis was difficult to differentiate from that of streptococcal disease; 20 of the patients had originally been given antibiotics. In 68% of patients without complications, the serum concentration of C-reactive protein was less than 20 mg/L, suggesting a viral disease. No other hematologic test was found helpful in identifying the adenoviral tonsillitis. The rapid detection of adenovirus antigen permitted withdrawal of unnecessary and ineffective antibiotic treatment in most patients.

Adenoviridae Infections↗