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PubMed2002

Heterogeneity of freshly isolated human tonsil dendritic cells demonstrated by intracellular markers, phagocytosis, and membrane dye transfer.

BACKGROUND: Heterogeneity within human dendritic cells (DCs) has been described but its functional relationships to cells of macrophage lineage and its role in human immunodeficiency virus (HIV) infection in vivo remain unclear. METHODS: Tonsil macrophages and DCs were isolated from low-density cells by negative selection and DCs were sorted into myeloid and plasmacytoid populations using antibodies to CD11c or CD123. Phagocytosis of latex beads and uptake of dye-labeled target cells were compared by flow cytometry and CD68 and S-100 by immunofluorescence on cytospins of sorted cells. RESULTS: Bead uptake and membrane dye transfer were found in both blood and tonsil CD11c(+) DCs and in CD14(+) cells particularly from blood monocytes. CD11c(-) DCs were poorly phagocytic but took up fluorescent dye from intact, necrotic or apoptotic cells. Tonsil DCs and macrophages expressed both CD68 and S-100 but CD11c(-) DCs expressed CD68 only. CONCLUSIONS: Freshly isolated CD11c(+) tonsil DCs are similar to CD14(+) macrophages in phagocytic function but the poorly phagocytic CD11c(-) DCs can also take up membrane from target cells. The intracellular markers commonly used to identify DCs and macrophages in situ do not identify accurately the CD11c(-) DC subset nor do they distinguish tonsil macrophages from DCs.

Antigens, CD↗
PubMed1987

Analysis of the human tonsil B cell repertoire by somatic hybridization: occurrence of both "monospecific" and "multispecific" (auto)antibody-secreting cells.

The purpose of our study was to investigate the (auto)antibody repertoire of human tonsil B cells. To this end, unseparated tonsil B cells and "large" and "small" tonsil B cells, obtained after fractionation on a discontinuous Percoll gradient, were fused with the (human X mouse) heterohybridoma K6H6/B5. The IgM antibodies secreted by individual clones were analyzed in spot-enzyme-linked immunosorbent assay for reactivity against a panel of 5 autoantigens and 1 xenoantigen. The results show that both "mono-" and "multispecific" (auto)antibody-secreting clones can be obtained by somatic hybridization of unseparated, "small" and "large" tonsil B cells. The patterns of IgM reactivities in these three collections of hybridomas did not differ significantly. Inhibition experiments revealed that "multispecific" antibodies can bind with high affinity to more than one (auto)antigen. The results of these experiments are discussed in relation to the state of activation of tonsil B lymphocytes.

Antibody Specificity↗
PubMed2006

Isolation and characterization of lymphatic microvascular endothelial cells from human tonsils.

Human lymphatic endothelial cells (LECs) have isolated prevalently from human derma and tumors. As specialized lymphatic organs within the oropharynx, palatine tonsils are easily obtained and rich in lymphatic venules. Using a two-step purification method based on the sorting of endothelial cells with Ulex Europaeus Agglutinin 1 (UEA-1)-coated beads, followed by purification with monoclonal antibody D2-40, we successfully purified LECs from human palatine tonsils. The LECs were expanded on flasks coated with collagen type 1 and fibronectin for up to 8-10 passages and then analyzed for phenotypic and functional properties. Cultured cells retained the phenotypic pattern of the lymphatic endothelium of palatine tonsils and expressed functional VEGFR-3 molecules. In fact, stimulation with VEGFR-3 ligand, the vascular endothelium grow factor C, induced a marked increase in cell proliferation. Similarly to blood endothelial cells (BECs), LECs were able to form tube-like structure when seeded in Cultrex basement membrane extract. Comparative studies performed on LECs derived from palatine tonsils and iliac lymphatic vessels (ILVs), obtained with the same procedures, showed substantial discrepancies in the expression of various lymphatic markers. This points to the existence of micro- and macrovessel-derived LECs with different phenotypes, possibly involving different biological activities and functions. Palatine tonsil- and ILV-derived LECs may, therefore, represent new models for investigating function and biochemical properties of these lymphatic endothelia.

Antibodies, Monoclonal↗
PubMed1982

The role of BK virus in acute respiratory tract disease and the presence of BKV DNA in tonsils.

The significance of BKV infections relative to infections by generally tested respiratory agents was investigated in children with acute respiratory disease. Paired sera from 177 children admitted to a hospital for acute respiratory disease were tested for significant rises in antibodies. Sera from seven patients showed a seroconversion to BKV and clinical signs of acute upper respiratory tract infection were exhibited by each of these patients. BKV infections were present in 8% of the patients with upper respiratory tract disease while seroconversions to adenovirus (2%), influenza A virus (1%), parainfluenza virus (5%), RS virus (6%) and mycoplasma pneumoniae (1%) were observed in 15% of the patients with upper respiratory tract disease. BKV was isolated from the urine of one child with tonsillitis with a concomitant seroconversion to BKV. Tonsils from children with recurrent attacks of acute respiratory disease were tested for the presence of BKV DNA by hybridization with a cloned genomic 32P-labeled DNA of prototype BKV. Five of twelve tonsil DNAs showed hybridization with BKV DNA. Each tonsil showing hybridization with BKV DNA contained multiple nonintegrated copies of the BKV genome per diploid amount of host cell DNA. Attempts to recover infective BKV by transfection of primary human embryonic cells with tonsil DNAs or by co-cultivation of tonsillar cells with primary human embryonic cells were unsuccessful.

Acute Disease↗
PubMed1981

The enzyme histochemistry of lymphoid and non-lymphoid cells of the human palatine tonsil: a basis for the study of lymphomas.

The distribution of various hydrolytic enzymes has been determined in 27 human palatine tonsils by means of conventional enzyme histochemical techniques, and lysozyme (muramidase) activity has been localised in eight tonsils by the unlabelled antibody peroxidase-antiperoxidase complex (PAP) method. The enzyme activities of various cell types are compared and the effect of various methods of fixation and processing discussed. The results suggest that arrangement of various histiocytic cells types within the tonsillar follicles and crypt epithelium is related to the processing of antigen. The PAP method for lysozyme demonstrates a smaller population of cells than is demonstrated by the alpha-naphthyl acetate esterase (ANAE) method. T-cells are demonstrated by the presence of dot-like ANAE activity in their cytoplasm. Large numbers of the lymphocytes of this type were located in the T-dependent areas of the tonsil, and are frequent beneath the crypt epithelium. The efferent lymphatic vessels appeared to contain an almost pure population of T-cells. The immunohistochemical method for lysozyme did not differentiate between T- and B-cell areas, dot-like activity being absent. As in other workers' studies on non-lymphoid cells of the murine spleen, several types of glass-adherent cells have been identified in short-term cell cultures from the human tonsil. True dendritic cells and branching macrophages differ in several ways (as in the mouse spleen). The tonsil is considered to be a useful control "reactive" lymphoid organ, to act as a baseline tissue in an extended study of morphology and enzyme histochemistry in the lymphomas.

Adolescent↗
PubMed1977

[Concentrations in serum and tonsils after infection of azidocillin (author's transl)].

Prior to tonsillectomy 57 selected patients were given 1 g azidocillin i.v. in the form of a 10% aqueous solution. Serum samples were drawn at different times after the injection and both the serum and the tonsils were examined for azidocillin activity. The results of examination of 133 serum samples and 111 tonsils were evaluated, in addition to 74 paired values for tonsils and serum. Serum levels attained a maximum of 230 microng/ml and remained within measurable range for at least six hours. Tonsil levels reached a maximum of 17 microng/g and after three hours began to drop below measurable range. The quotient for distribution between tonsils and serum rose during the first hour to 20-25% (average approx. 10%) and was about 3-6% three to four hours after injection.

Adult↗
PubMed1984

Analysis of lymphoid and dendritic cells in human lymph node, tonsil and spleen. A study using monoclonal and heterologous antibodies.

The distribution of lymphoid and dendritic cells in human reactive lymph nodes, tonsils and spleens was examined by means of an indirect immunoperoxidase technique, using a panel of monoclonal and heterologous antibodies. The antibodies used were directed against antigens present on T cell subsets (Leu1, leu2a, Leu3a, TA1, OKT6), various types of B cells (BA1, BA2, HLA-DR, CR1) and cells of the mononuclear phagocyte system (alpha HM1, TA1, CR1, OKM1, NA 1/34). In the lymph node and tonsil Leu3a-positive cells (T-helper/inducer phenotype) and Leu2a-positive cells (T-suppressor/cytotoxic phenotype) are found in the thymus-dependent or T-cell area; in the spleen Leu3a-positive cells are found mostly in the periarteriolar lymphocyte sheath (PALS), while Leu2a-positive T-suppressor/cytotoxic cells are almost completely restricted to the cords of Billroth in the red pulp. The cells in the mantle zone of germinal centres and in the primary follicles in lymph nodes, tonsils and spleens have B-cell properties (BA1-, HLA-DR-, and CR1-positive). The cells in the germinal centres show a similar staining pattern (HLA-DR-, and partly CR1-positive). Follicles and T-cell-dependent areas have specific dendritic cells, each with a specific staining pattern: the dendritic reticulum cell (DRC) of the follicle stain with CR1, HLA-DR, BA2 and alpha HM1; the interdigitating cell of the T-cell areas in the lymph node, tonsil and spleen stain with HLA-DR and BA1. Moreover, large dendritic OKT6-positive cells are found in the T-cell areas of some of the peripheral lymph nodes, and are probably Langerhans cells. It is concluded that human lymph nodes and tonsils have an identical compartimentalisation, clearly differing from the spleen in cellular organization.

Antibodies, Monoclonal↗
PubMed1981

Characteristic distribution of immunoglobulin-containing cells in palatine tonsils.

Using fluorescein-labelled antibodies against gamma, mu and alpha chains, Ig-containing cells in palatine tonsils were studied in 120 patients. The aim of this study was to determine the most frequently repeated typical findings as regards the numbers and localisation of these cells in tonsils and to confront the data obtained with the concept that tonsils are a component of the local immunity system. The preponderance of IgG over IgA cells was confirmed, both cell types being preferentially localized in extrafollicular tissue whereas IgM was mostly found in germinal centres. Together with progressing tonsillar atrophia, the frequency of positive findings of IgM decreased, whereas the numbers of IgG and IgA cells were proportional to the amount of remaining lymphoid tissue. IgA cells were not preponderant in tonsils and their localization in the surface layer of epithelium was rather exceptional, SC antigen could not be demonstrated unequivocally and the morphological picture in germinal centres was characteristic for IgM production rather for IgA. Thus the palatine tonsils according to the content and distribution of immunocytes, correspond to the lymph node rather than to an organ involved significantly in the local antibody formation.

Adolescent↗
PubMed2004

The role of oxidants and antioxidants in chronic tonsillitis and adenoid hypertrophy in children.

OBJECTIVE: To determine the possible role of oxidants and antioxidants in the pathogenesis of chronic tonsillitis (CT) and adenoid hypertrophy (AH) in children. METHODS: Randomized, prospective, controlled. The study group was made up of children with chronic tonsillitis and adenoid hypertrophy who are to undergo tonsillectomy and adenoidectomy. The control group was constructed with otherwise healthy children with normal ENT examination. The blood levels of antioxidants (retinol, beta-carotene, alpha-tocopherol, laycopene, ascorbic acid, superoxide dismutase, glutathione peroxidase, GSH) and peroxidation products (malondialdehyde) were determined before and 1 month after the operation in the study group and once only in the control group. These antioxidants and peroxidation products were also measured in the tonsil and adenoid tissue that were obtained during operation. RESULTS: In the study group, the blood levels of antioxidants and oxidant before and after the operation were significantly different when compared to the control group (P < 0.05). In the study group, the blood antioxidant levels increased and oxidant level decreased significantly after the operation (P < 0.05). These levels after the operation never reached those of the control group. CONCLUSIONS: Oxidants and antioxidants played a significant role in the pathogenesis of chronic tonsillitis and adenoid hypertrophy in children. These children are under significant oxidative stress. Tonsillectomy and adenoidectomy significantly decreased the oxidative stress in these patients, but could not normalize it completely. Further studies are necessary to evaluate their possible therapeutic role in preventing recurrent tonsillitis and treating postoperative patients to help normalize their blood levels of antioxidants.

Adenoidectomy↗
PubMed2004

Investigation of antioxidant status in children with acute otitis media and tonsillitis.

OBJECTIVE: Antioxidant enzymes status was investigated in children with acute otitis media (AOM) and acute tonsillitis. The aim was to determine the effect of oxidative damage due to free radicals on the antioxidant enzymes status in children with AOM and acute tonsillitis. METHODS: We measured erythrocyte levels of superoxide dismutase and glutathione peroxidase (GPx) and activity of serum catalase, ceruloplasmin, albumin and total bilirubin in 23 children with AOM, 27 with acute tonsillitis and 17 control subjects. RESULTS: There was statistically significant difference between the groups for all parameters. All antioxidant activities except for albumin level were increased in the studies groups. When the values of AOM and acute tonsillitis were compared, there was statistically significant difference between the groups for erythrocyte GPx. CONCLUSION: Our findings indicate that not only a few antioxidant enzymes but almost all antioxidant activities showed a significant enhancement in children with AOM and acute tonsillitis.

Acute Disease↗
PubMed2005

Detection of herpesvirus type 8 (HHV8) in children's tonsils and adenoids by immunohistochemistry and in situ hybridization.

OBJECTIVE: Human herpesvirus 8 (HHV8) has been associated with multicentric Castleman's disease, Kaposi's sarcoma and effusion non-Hodgkin's lymphoma. Epidemiological studies have shown seropositivity in variable proportions of populations. It seems to be sexually transmitted among adults and through oral contact among children. The virus has been demonstrated in desquamating oral epithelial cells, but there is no report on its presence in the Waldeyer's ring. The purpose of the present study is to detect HHV8 in tonsils and adenoids from children up to 20 years of age in which these organs had been surgically removed due to hypertrophy, using immunohistochemistry and in situ hybridization. METHODS: Paraffin wax-embedded sections consisting of 181 tonsils and 162 adenoids from 293 patients were analyzed. HHV8 was detected by immunohistochemistry (IHC) using the anti-LNA1 antibody (Novocastra) and the LSAB+ detection system (Dako). For the in situ hybridization (ISH), the T1-1 probe for the viral mRNA and the detection system used were provided by Novocastra. RESULTS: In 20 cases (6.83%), HHV8 was detected in cells morphologically characterized as lymphoid. In three of them epithelial cells were also positive. In 19 cases, the virus was detected in tonsils and in just 1 case in an adenoid. In all 20 cases detection was possible by ISH, whereas in only 2 of them there was a concomitant positivity by IHC. CONCLUSION: Our data support the oral route of contamination by HHV8 in children, in whom tonsils and adenoids may harbor the virus. It is found especially in tonsils and only rarely in adenoids. In these organs, ISH is the method of choice to detect this virus, probably due to the small amount of viral proteins.

Adenoids↗
PubMed2001

Phenotypic characterization of five dendritic cell subsets in human tonsils.

Heterogeneous expression of several antigens on the three currently defined tonsil dendritic cell (DC) subsets encouraged us to re-examine tonsil DCs using a new method that minimized DC differentiation and activation during their preparation. Three-color flow cytometry and dual-color immunohistology was used in conjunction with an extensive panel of antibodies to relevant DC-related antigens to analyze lin(-) HLA-DR(+) tonsil DCs. Here we identify, quantify, and locate five tonsil DC subsets based on their relative expression of the HLA-DR, CD11c, CD13, and CD123 antigens. In situ localization identified four of these DC subsets as distinct interdigitating DC populations. These included three new interdigitating DC subsets defined as HLA-DR(hi) CD11c(+) DCs, HLA-DR(mod) CD11c(+) CD13(+) DCs, and HLA-DR(mod) CD11c(-) CD123(-) DCs, as well as the plasmacytoid DCs (HLA-DR(mod) CD11c(-) CD123(+)). These subsets differed in their expression of DC-associated differentiation/activation antigens and co-stimulator molecules including CD83, CMRF-44, CMRF-56, 2-7, CD86, and 4-1BB ligand. The fifth HLA-DR(mod) CD11c(+) DC subset was identified as germinal center DCs, but contrary to previous reports they are redefined as lacking the CD13 antigen. The definition and extensive phenotypic analysis of these five DC subsets in human tonsil extends our understanding of the complexity of DC biology.

CD13 Antigens↗
PubMed2000

Human nasopharyngeal-associated lymphoreticular tissues. Functional analysis of subepithelial and intraepithelial B and T cells from adenoids and tonsils.

Subepithelial and intraepithelial lymphocytes of human adenoids and tonsils were characterized and directly compared to determine the potential contribution of these tissues to mucosal and systemic immune responses. The distribution of T and B cell subsets, cytokine patterns, and antibody (Ab) isotype profiles were similar for adenoids and tonsils. Both tissues contained predominantly B cells ( approximately 65%), approximately 5% macrophages, and 30% CD3(+) T cells. The T cells were primarily of the CD4(+) subset ( approximately 80%). Tonsillar intraepithelial lymphocytes were also enriched in B cells. The analysis of dispersed cells revealed a higher frequency of cells secreting IgG than IgA and the predominant Ig subclass profiles were IgG1 > IgG3 and IgA1 > IgA2, respectively. In situ analysis also revealed higher numbers of IgG- than IgA-positive cells. These IgG-positive cells were present in the epithelium and in the subepithelial zones of both tonsils and adenoids. Mitogen-triggered T cells from tonsils and adenoids produced both Th1- and Th2-type cytokines, clearly exhibiting their pluripotentiality for support of cell-mediated and Ab responses. Interestingly, antigen-specific T cells produced interferon-gamma and lower levels of interleukin-5. These results suggest that adenoids and tonsils of the nasopharyngeal-associated lymphoreticular tissues represent a distinct component of the mucosal-associated lymphoreticular tissues with features of both systemic and mucosal compartments.

Adenoids↗
PubMed1994

Herpes simplex infection causing acute necrotizing tonsillitis.

OBJECTIVE: To describe the clinical and pathologic features of acute herpetic tonsillitis and to compare the histologic findings with those of herpetic lymphadenitis. DESIGN: We present a case report of a 22-year-old woman with bilateral cervical adenopathy, acute tonsillitis, and suspected peritonsillar abscess. MATERIAL AND METHODS: Histologic examination of the excised tonsils demonstrated discrete necrotic areas that contained cells with intranuclear viral inclusions. RESULTS: The diagnosis of herpetic tonsillitis was confirmed by demonstrating herpes simplex virus (HSV)-infected cells on paraffin section immunostains and by positive HSV cultures of the tonsillar tissue. CONCLUSION: HSV infection is an uncommon cause of acute tonsillitis; the histologic findings are similar to those seen in herpes simplex lymphadenitis.

Acute Disease↗
PubMed2000

Human adult tonsil xenotransplantation into SCID mice for studying human immune responses and B cell lymphomagenesis.

OBJECTIVE: To generate a human-mouse xenochimeric model where human cells remain clustered in the animal to optimize their interactions and recovery. MATERIALS AND METHODS: Severe combined immune deficient mice (SCID) were xenografted subcutaneously with human adult tonsil pieces (hu-ton-SCID mice). Such animals were: (a) compared with those receiving tonsil cells in suspension, and (b) immunized with de novo and recall antigens. RESULTS: Human tonsil pieces survived a long period of time in SCID mice, while polyclonal human T- and B-lymphocytes persisted in close vicinity within the implantation area; however, little or no graft-versus-host disease was detectable. Not surprisingly, local development of lymphoproliferative disease was often observed in animals receiving lymphoid implants from donors previously infected by the Epstein-Barr virus. One month after surgery, higher serum levels of human IgG were found in SCID mice transplanted with tonsil pieces (2x10(7) cells/animal) than in animals injected with 5x10(7) tonsil cells in suspension (1.9 vs. 0.3 mg/mL, p < 0.002). Importantly, the production of human IgG in hu-ton-SCID mice remained polyclonal for at least 6 months and was linked to the presence of cells within the implants. Immunization of hu-ton-SCID mice with hepatitis B core, a de novo antigen, did not produce a significant IgG immune response; however immunization with tetanus toxoid (TT), a thymus-dependent recall antigen, yielded high (> 700-fold increase in anti-TT IgG levels) and long-lasting (> 6 months) secondary immune responses. CONCLUSION: The hu-ton-SCID mouse xenochimeric model described in this report may improve our understanding of human lymphoid cell interactions, secondary immune responses, and lymphomagenesis.

Adult↗
PubMed1999

The 7alpha-hydroxysteroids produced in human tonsils enhance the immune response to tetanus toxoid and Bordetella pertussis antigens.

Human tonsils were assessed for their ability to 7alpha-hydroxylate pregnenolone (PREG), dehydroepiandrosterone (DHEA) and 3-epiandrosterone (EPIA). Both 7alpha-hydroxy-DHEA and 7alpha-hydroxy-EPIA were produced by homogenates of either whole tonsils or of lymphocyte-depleted tonsil fractions. In contrast, isolated lymphocytes were found to be unable to carry out 7alpha-hydroxylation. When co-cultures of tonsil-derived T and B lymphocytes were set up under stimulatory conditions, IgGs were released in the supernatants and could be quantitated, and immunomodulating properties of different steroids were monitored. When PREG was added to a mixture of tonsil-derived B and T lymphocytes, a decrease of non-specific and specific IgG was observed. An increase in specific anti-tetanus toxoid and anti-Bordetella pertussis antigen IgGs was obtained with either 1 microM 7alpha-hydroxy-DHEA or 1 microM 7alpha-hydroxy-EPIA. In contrast, DHEA and EPIA were unable to trigger such an effect. When cultures of isolated tonsillar B cells were used, none of the steroids tested showed significant effects on specific IgG productions. These data led to the conclusion that human tonsillar cells transform DHEA and EPIA, but not PREG, into 7alpha-hydroxylated metabolites. These metabolites could act on target tonsillar T lymphocytes which in turn act upon B lymphocytes for increasing specific IgG production.

Adolescent↗
PubMed1986

Immunological reaction to horseradish peroxidase of the palatine tonsil of the rabbit.

Detection of the immune response to external antigen of the palatine tonsil of the rabbit is an interesting problem. Employing horseradish peroxidase (HRP) as an external antigen, the reactions of a perfusion sensitized group and those of a group sensitized by intravenous injection were examined. The results were as follows: 1. Hypertrophy of the palatine tonsil was recognized only in the perfusion sensitized group. 2. The serum titer level of the antibody to HRP was recognized to be high in the intravenous injection sensitized group, which was confirmed by the production of antibody in the spleen. It was also confirmed in the repeated perfusion sensitized group at a low level. 3. Antigen invasion through the epithelium of the crypt was possible by repeated intravenous injection sensitization as for perfusion sensitization. Also, invading HRP was observed electron microscopically to be phagocytized by macrophages and dendritic reticular cells. 4. In the perfusion sensitized group, production of antibody was not found in the palatine tonsils but in the regional lymph nodes of the neck. 5. In the perfusion sensitized group, a large number of eosinophils were noted mainly on the basal side of the interfollicular zone. 6. In the intravenous injection sensitized group, anti-HRP was found in the palatine tonsils and also in the spleen. Participation of the tonsils has been revealed in general human immunity.

Animals↗
PubMed2003

Blood vascular architecture of the palatine tonsil in the musk shrew (Suncus murinus): scanning electron microscopic study of corrosion casts.

The musk shrew, Suncus murinus, is one of the primitive mammals and has a pair of palatine tonsils. In the present study, we investigated the blood microvascular architecture of the tonsil in this animal by scanning electron microscopy of corrosion casts. The paranodular arterioles entered the lymph nodule to form a coarse capillary plexus within the nodule. Some of the arterioles reached the dome region to give rise to a fine meshwork of dome subepithelial capillaries. This dome subepithelial capillary network did not show any hairpin or switch-back patterns, as seen in human and rabbit tonsils. Both of the nodular and dome capillaries were drained into the postcapillary venules in the periphery of the nodular or the paranodular region. On the surface of these cast venules, oval-shaped indentations were seen corresponding to the luminal surface of the high endothelial venules. These venules were collected into the large vein at the bottom of the tonsil. The blood vascular architecture of the musk shrew tonsil is basically the same as those of other mucosa-associated lymphoid tissues in mammals.

Animals↗