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PubMed1993

Characterization of cytokine production in infectious mononucleosis studied at a single-cell level in tonsil and peripheral blood.

Cytokine profile and production was studied at a single-cell level in cells obtained from 14 patients with acute infectious mononucleosis (IM), with less than 7 days of symptomatic disease, by use of cytokine-specific MoAbs and indirect immunofluorescence technique. In producer cells, all the studied cytokines, except IL-1, accumulated in the Golgi system, which resulted in a characteristic morphology of the staining. Less than one in a thousand mononuclear cells obtained directly from IM blood and stained within 2 h of sampling produced IL-2, interferon-gamma (IFN-gamma), IL-4, IL-5, IL-6, IL-10, GM-CSF, tumour necrosis factor-alpha (TNF-alpha) or TNF-beta, spontaneously. However, these cells were induced to cytokine synthesis by T cell receptor ligation in vitro using immobilized anti-CD3 MoAbs for 2-3 h restimulation under conditions which did not activate normal cells. By this approach 168 +/- 120 cells/10,000 peripheral blood mononuclear cells produced IFN-gamma as compared with 10 +/- 8 cells/10,000 non-stimulated cultured cells obtained from IM patients (P < 0.001) and 1/10,000 cells obtained from healthy controls, respectively. No induced production of IL-2, IL-3, IL-4, IL-5, IL-10, GM-CSF or TNF-beta was detected in IM cells obtained from peripheral blood by this restimulation. In contrast, a spontaneous cytokine production was evident in tonsil material obtained from four IM patients tonsilectomized because of respiratory obstruction. From this site 160 +/- 40 cells/10,000 cells produced IL-2, 40 +/- 30 cells IL-6, 30 +/- 30 cells TNF-beta and 35 +/- 25 cells IFN-gamma, respectively. No such spontaneous IL-2, IL-6, TNF-beta or IFN-gamma production was evident in control cells obtained from patients tonsilectomized because of chronic tonsil hyperplasia.

Adolescent↗
PubMed1986

Dendritic reticulum cells in reactive lymph nodes and tonsils: an immunohistological study.

There has recently been much interest in the patterns of follicular dendritic reticulum cells (DRC) in pathological lymph nodes, particularly in relation to the phenomenon of DRC break-up (thought to be pathognomonic of AIDS-related lymphadenopathies) and to progressive transformation of germinal centres (as a possible precursor of lymphocyte predominant Hodgkin's disease). In the present study we have immunostained twenty-nine reactive lymph nodes and five tonsils with monoclonal antibody R4/23 (DAKO-DRC) in order to evaluate the frequency of such changes in lymphoid tissue unaffected by AIDS or Hodgkin's disease. Most of the specimens contained typical secondary follicles with clearly defined germinal centres and mantle zones. There were two variants in lymph nodes showing follicular hyperplasia characterized by (i) progressive transformation of germinal centres and (ii) inclusions of nests of small lymphocytes within germinal centres. In each of these types of follicles the compact evenly-distributed meshwork of DRCs, as previously described, was seen. However there were considerable variations in DRC meshwork in each category (the pattern could not be predicted from the morphology) with examples in all three of the DRC break-up previously considered specific for the AIDS related lymphadenopathy. Since none of the lymph nodes and tonsils studied had any known relationship to either Hodgkin's disease or AIDS it is argued that none of the changes in the DRC meshwork observed are specific for these conditions.

Acquired Immunodeficiency Syndrome↗
PubMed1981

Human membrane-bound C3 receptors. II. Physiochemical properties of the C3b and C3d receptors isolated from tonsil cells by immunoprecipitation.

The aim of the present study was to define the physicochemical structure of C3b and C3d receptors of lymphoid cells. C3b and C3d receptors were isolated from KBr lysates of the 20,000 g fraction of human tonsil homogenates by immunoprecipitation with an anti-C3 receptor serum (AC3RS). Sodium dodecyl sulphate (SDS) gel filtration and polyacrylamide gel electrophoresis (PAGE) of unreduced immunoprecipitates revealed a highly predominant component with an apparent molecular weight (mol. wt.) greater than 1 x 10(6) and a small component with a mol. wt. of 80,000. After reduction, the SDS-PAGE profile was made up of a constant major 38,000 mol. wt. component and a inconstant smaller 18,000 mol. wt. component. The 38,000 (and also the 18,000) component could be isolated only from C3 receptor-active lysates, and not from C3 receptor-negative lysates. Taken together, the results of this study suggest that the active C3 receptor molecule of tonsil cells is a lipoprotein complex with a mol. wt. greater than 1 x 10(6); its protein moiety consists predominantly of disulphide-bridged polypeptide chains with a mol. wt. of 38,000; C3b and C3d receptors are composed of equal-sized polypeptide chains, but the specific binding sites for C3b and C3d are located on different molecules.

Chromatography, Gel↗
PubMed2006

Diversity of the Ig repertoire is maintained with age in spite of reduced germinal centre cells in human tonsil lymphoid tissue.

Humans and almost all species studied to date exhibit a decreased responsiveness to immunization and increased autoimmunity with age. While this has been observed clinically for decades, only recently has an understanding of the molecular basis for these changes begun to be appreciated. Studies of the B-cell aspects of these changes in ageing mice and the very few reports in ageing humans have not been conclusive. Here we examine the nucleotide sequence of over 1250 VH transcripts from the tonsils of individuals of various ages for changes to the VH4 immunoglobulin repertoire. An exhaustive examination of VH, DH and JH gene segment utilization revealed a remarkable similarity of the repertoires. The extent of somatic hypermutation was fully maintained or even increased by some measures into the eighth decade of life. However, we found by middle age that the representation of naïve and germinal centre B-cell subpopulations changed relative to total B lymphocytes in the tonsil. While the percentage of naïve and germinal centre B-cell subpopulations changes during the second half of life, these findings suggest that even with advancing age, humans remain capable of generating an extremely diverse Ig repertoire while maintaining a similar spectrum of Ig rearrangements once the germinal centre reaction begins.

Adult↗
PubMed1990

Expression of T-cell receptor (TCR)alpha chain on normal human tonsils and T-cell lymphomas.

We analyzed immunohistologically the expression of T-cell receptor (TCR)alpha chain on human tonsils and on T-cell lymphoma (T-ML) tissues using the avidin-biotin-peroxidase method. A murine monoclonal antibody alpha F1, specific for the constant region of the TCR alpha chain, was employed. On normal tonsil, alpha F1-positive cells were observed mainly in T-zones and germinal centers. In T-zones, the staining intensities varied markedly, with heavy staining evident in less than one fourth. In germinal centers, a proportion of stained cells showed a histiocytic pattern with small cytoplasmic projections. All the T-ML tissues expressed TCR alpha, whereas the staining intensities varied among cases and among lymphoma cells. No correlations were observed in the expressions of TCR alpha chain and other T-cell markers including CD3, CD4 and CD8.

Antigens, Differentiation, T-Lymphocyte↗
PubMed1980

The invitro response of lymphocytes from adenoid vegetations and tonsils to PPD. Influence of autologous blood monocytes, T lymphocytes and unseparated lymphocytes.

Thymidine incorporation in lymphocytes obtained from adenoids (AVL), tonsils (TL) and blood (PBL) and stimulated by PPD was compared in 19 BCG-vaccinated patients. Responses were approximately tenfold higher in vaccinated than in unvaccinated patients. The responses obtained for AVL/TL were lower than for PBL in BCG-vaccinated as well as in unvaccinated patients. In autologous mixtures of AVL/TL and PBL a linear increase in thymidine incorporation was obtained with increasing numbers of PBL. The monocyte concentration in cell suspensions obtained from adenoid and tonsil tissue was lower than in cells prepared from blood; addition of autologous monocytes purified from blood to cultures of AVL and TL was followed by a small increase in response to PPD, but not to the level of responsiveness of PBL. Addition of autologous T lymphocytes purified from blood to cultues of AVL and TL was followed by a small increase in thymidine incorporation, of similar proportions in cultures with and without PPD. It is concluded that the proportion of PPD-sensitized lymphocytes is smaller in AVL/TL than in PBL.

Adenoids↗
PubMed1991

[Recovery of Chlamydia pneumoniae and Chlamydia trachomatis in a patient with recurrent tonsillitis, bronchitis and otitis media with effusion].

We report a case of recurrent tonsillitis and otitis media with effusion (OME) from which Chlamydia trachomatis was isolated. Chlamydia pneumoniae, a newly recognized species of Chlamydia, was also recovered from the tonsillar and bronchial swabs. A 8-year-old girl was seen on February 23, 1988, because of a running nose, a productive cough and bilateral hearing difficulty. She had a history of recurrent tonsillitis. The diagnosis was acute sinusitis with tubal obstruction, then cefixime was prescribed. Her symptoms were once resolved, for the time being but she came back to the hospital a week later with a bilateral ear-ache. The tympanic membranes were injected and characteristically retracted. Her left ear showed type B tympanogram (effusion). Tympanocentesis was performed to remove middle-ear effusion, from which C. trachomatis but no ordinary bacterium was isolated. Therefore rokitamycin 300 mg/day was administered for a week. Her condition improved, however, a rhinorrhea, a plugged ear sensation and a hacking cough returned in a month. She was admitted to the hospital on May 10, for tympanostomy and grommet insertion, but from the day before admission, she had a sore throat with fever (39.2 degrees C). The surgery was withheld until May 26. When adenotonsillectomy and grommets insertion were undertaken, C. trachomatis had disappeared from the middle-ear effusion, but C. pneumoniae was recovered from both tonsillar and bronchial swabs. Readministration of rokitamycin was performed and to date (June, 1990) she remains well.

Bronchitis↗
PubMed1979

Cell-mediated immune response to bacterial products in human tonsils and peripheral blood lymphocytes.

Lymphoproliferative responses of tonsillar tissue lymphocytes and peripheral blood lymphocytes to phytohemagglutinin and specific bacterial product antigens were studied in children undergoing tonsillectomy and adenoidectomy. Tonsillar tissue lymphocytes responded to optimal concentrations of phytohemagglutinin. Varidase, and streptolysin-O in a manner similar to peripheral blood lymphocytes. Higher base-line mitogenic activity in tonsillar lymphocytes was frequently associated with the presence of Staphylococcus aureus in the tonsils. Tonsillar tissue lymphocytes from 23% of the subjects with the highest base-line mitogenic activity manifested a decreased response to in vitro stimulation with mitogens or antigens. In subjects with such preactivated tonsillar lymphocytes, the proliferative responsiveness of blood lymphocytes to mitogen and antigens was markedly increased after tonsillectomy and adenoidectomy. These observations suggest the existence of in vitro correlates of cellular immunity to bacterial products in the mucosal surfaces. In addition, it is proposed that tonsils may possess immunosuppressive activity for peripheral blood lymphocytes, which may be related to local tonsillar infections.

Adenoidectomy↗
PubMed1998

Development of an immunomagnetic method for selective isolation of Actinobacillus pleuropneumoniae serotype 1 from tonsils.

An immunomagnetic separation technique (IMS) for the selective isolation of Actinobacillus pleuropneumoniae serotype 1 was developed. Superparamagnetic polystyrene beads (immunomagnetic beads [IMBs]) were coated with purified rabbit immunoglobulin G specific for A. pleuropneumoniae serotype 1. The antibody concentration, the number of IMBs, the incubation time, and the temperature of incubation influenced the recovery of the target bacteria. The sensitivity of the IMS technique was 1,000-fold higher than that of direct culture. When tonsils from animals from infected herds were tested, significantly more positive tonsils were detected by the IMS technique (68%) than by the standard procedures (22%). The method represents an innovative and highly sensitive approach for the isolation of A. pleuropneumoniae from carrier animals.

Actinobacillus pleuropneumoniae↗
PubMed2002

The simian immunodeficiency virus deltaNef vaccine, after application to the tonsils of Rhesus macaques, replicates primarily within CD4(+) T cells and elicits a local perforin-positive CD8(+) T-cell response.

Deletion of the nef gene from simian immunodeficiency virus (SIV) strain SIVmac239 yields a virus that undergoes attenuated growth in rhesus macaques and offers substantial protection against a subsequent challenge with some SIV wild-type viruses. We used a recently described model to identify sites in which the SIVDeltanef vaccine strain replicates and elicits immunity in vivo. A high dose of SIVDeltanef was applied to the palatine and lingual tonsils, where it replicated vigorously in this portal of entry at 7 days. Within 2 weeks, the virus had spread and was replicating actively in axillary lymph nodes, primarily in extrafollicular T-cell-rich regions but also in germinal centers. At this time, large numbers of perforin-positive cells, both CD8(+) T cells and CD3-negative presumptive natural killer cells, were found in the tonsil and axillary lymph nodes. The number of infected cells and perforin-positive cells then fell. When autopsy studies were carried out at 26 weeks, only 1 to 3 cells hybridized for viral RNA per section of lymphoid tissue. Nevertheless, infected cells were detected chronically in most lymphoid organs, where the titers of infectious virus could exceed by a log or more the titers in blood. Immunocytochemical labeling at the early active stages of infection showed that cells expressing SIVDeltanef RNA were CD4(+) T lymphocytes. A majority of infected cells were not in the active cell cycle, since 60 to 70% of the RNA-positive cells in tissue sections lacked the Ki-67 cell cycle antigen, and both Ki-67-positive and -negative cells had similar grain counts for viral RNA. Macrophages and dendritic cells, identified with a panel of monoclonal antibodies to these cells, were rarely infected. We conclude that the attenuated growth and protection observed with the SIVDeltanef vaccine strain does not require that the virus shift its characteristic site of replication, the CD4(+) T lymphocyte. In fact, this immunodeficiency virus can replicate actively in CD4(+) T cells prior to being contained by the host, at least in part by a strong killer cell response that is generated acutely in the infected lymph nodes.

AIDS Vaccines↗
PubMed2006

Spatial organisation of microbiota in quiescent adenoiditis and tonsillitis.

BACKGROUND: The reasons for recurrent adenotonsillitis are poorly understood. METHODS: The in situ composition of microbiota of nasal (5 children, 25 adults) and of hypertrophied adenoid and tonsillar tissue (50 children, 20 adults) was investigated using a broad range of fluorescent oligonucleotide probes targeted to bacterial rRNA. None of the patients had clinical signs of infection at the time of surgery. RESULTS: Multiple foci of ongoing purulent infections were found within hypertrophied adenoid and tonsillar tissue in 83% of patients, including islands and lawns of bacteria adherent to the epithelium, with concomitant marked inflammatory response, fissures filled with bacteria and pus, and diffuse infiltration of the tonsils by bacteria, microabscesses, and macrophages containing phagocytosed microorganisms. Haemophilusinfluenzae mainly diffusely infiltrated the tissue, Streptococcus and Bacteroides were typically found in fissures, and Fusobacteria,Pseudomonas and Burkholderia were exclusively located within adherent bacterial layers and infiltrates. The microbiota were always polymicrobial. CONCLUSIONS: Purulent processes persist during asymptomatic periods of adenotonsillitis. Most bacteria involved in this process are covered by a thick inflammatory infiltrate, are deeply invading, or are located within macrophages. The distribution of the bacteria within tonsils may be responsible for the failure of antibiotic treatment.

Abscess↗
PubMed1980

Effects of fixation and processing on immunohistochemical demonstration of immunoglobulin in paraffin sections of tonsil and bone marrow.

A number of fixatives were tested to determine their suitability for use with the unlabelled antibody peroxidase-antiperoxidase (PAP) method for demonstrating immunoglobulin in paraffin sections of tonsil and trephine samples of bone marrow. It was found that tonsil fixed in 'isotonic' solutions of formaldehyde reacted with the PAP method only after the sections had been trypsinised. Several other fixatives, including Bouin's fluid, Carnoy's fluid, and solutions containing mercuric chloride, gave tissues which reacted without trypsinisation of sections, and particularly good results were obtained with formol saline to which acetic acid (2-10%) had been added. A combination of acetic acid (10%)-formol saline and formol sublimate also gave excellent results with bone marrow. The influence on the PAP method of a number of steps in the processing of tissues and sections was also examined.

Bone Marrow↗
PubMed2001

New histochemical and ultrastructural observations on normal bovine tonsils.

Samples of normal bovine palatine tonsils were examined by light and electron microscopy. Like human tonsils, they were composed of crypts, subepithelial areas, follicles, and T-dependent zones, but their well-developed capsule subdivided the lymphoid tissue by connective septa. B cells formed the major lymphoid component. The follicles and T-dependent zones had morphological and histochemical features typical of peripheral lymph organs. Follicular dendritic cells were isolated and shown to be similar to human follicular dendritic cells.

Animals↗
PubMedDate not supplied

Effect of tonsillectomy on serum concentrations of interleukins and TNF-alpha in patients with chronic tonsillitis.

We investigated serum interleukin (IL)-1beta, IL-4, IL-6, IL-8 and tumor necrosis factor (TNF)-alpha levels in 17 children aged 5-12 years (mean 7) with chronic tonsillitis before and after tonsillectomy. Cytokine concentrations were measured by enzyme-linked immunosorbent assay. IL-1beta and IL-6 levels were significantly higher than the control levels (p < 0.05) in preoperative serum samples. Other cytokine levels were within normal limits. After tonsillectomy, IL-1beta and IL-6 levels were significantly reduced (p < 0.05). It is suggested that IL-1beta and IL-6 may be mediators which have a role in chronic tonsillitis disease and that tonsillectomy has a significant effect on preoperative IL-1beta and IL-6 serum levels.

Child↗
PubMed1992

Prothymosin alpha expression in lymph nodes and tonsils: an optical and ultrastructural study.

Prothymosin alpha (ProT alpha) distribution in human and rat lymph nodes and human tonsils was studied by means of immunohistochemical methods, using specific antibodies raised against thymosin alpha 1. We observed ProT alpha immunoreactivity in lymphoid cells of the germinal centers both in humans and rats. In human tonsils, positive cells were also seen in the basal layer of the surface epithelium. These results support the hypothesis that ProT alpha expression is restricted to actively proliferating cells. Immunoelectron microscopy revealed that ProT alpha was located in the nucleus, mainly in the border between euchromatin and heterochromatin.

Animals↗
PubMed1976

Increased proportion of B lymphocytes in human tonsils and appendices.

The percentage of T and B lymphocytes was studied in human tonsils and appendices. An increased percentage of B lymphocytes, 59.1% in tonsils and 54.5% in appendices, was found, whilst the percentage of T lymphocytes was 23.6% and 27.9%, respectively. In the peripheral blood of the corresponding figures were 12.5% for B cells and 50% for T cells.

Appendix↗
PubMed1976

Localization of E and EAC binding lymphocytes in human tonsils.

The localization of T and B lymphocytes in frozen sections of human tonsils was determined using E and EAC binding techniques. It was revealed that there were thymus-dependent areas with E binding lymphocytes in peri- and interfollicular zones. EAC binding lymphocytes were found mainly in the follicles. This suggests that B cells are produced in germinal centres. The present experiment confirmed that human tonsils contain two different kinds of lymphocytes demonstrating selective localizations in the tissues as similar to the lymph nodes.

Adolescent↗
PubMed1982

Amylase in the tonsil.

Excessive amounts of amylase were determined in 22 normal human tonsils by Winslow's method for the determination of serum amylase. The amylase content ranged from 44 to 704 U, mean 161 U. Since some malignant neoplasms are known to produce amylase, the present estimations might serve as a baseline for further investigation of lymphomas and carcinomas of the tonsils.

Adolescent↗