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Background: Many researchers have attempted to correlate p53 mutation and spontaneous apoptosis with the effectiveness of radiochemotherapy and with prognosis in several malignancies., Methods: The current study group consisted of 70 Japanese patients with maxillary sinus squamous cell carcinoma (SCC). Fifty seven patients were treated with radiochemotherapy followed by total or partial maxillectomy, and the remaining 13 patients were treated with radiotherapy alone. Tumor biopsy specimens at pretreatment status were examined for apoptosis-related proteins such as p53 protein, Fas, bax, bcl-x, and apoptosis using immunohistologic methods. The proportion of apoptotic cells labeled by single stranded DNA antibody was expressed as an apoptotic index (AI). p53 mutations at exons 5 through 8 were analyzed by direct sequence on polymerase chain reaction amplified products obtained from laser microdissected tissues. The effectiveness of radiochemotherapy was investigated histologically on surgically dissected specimens., Results: p53 mutations were identified in 20 (29%) of 70 patients. p53 protein was overexpressed in 39 patients (56%), Fas in 20 patients (29%), bax in 40 patients (57%), and bcl-x in 33 patients (47%). Overexpression of bax was associated with negativity of bcl-x (P = 0.015) and with high AI (P = 0.024). Low AI and/or p53 mutation in the pretreatment tissues correlated with low histologic effectiveness of radiochemotherapy (P = 0.048, P = 0.019, respectively). Kaplan-Meier analysis as well as univariate analysis using the Cox proportional hazards model showed that low histologic effectiveness of radiochemotherapy (P = 0.0281, P = 0.0284, respectively), p53 mutations (P = 0.0095, P = 0.0187, respectively), negativity of bax (P = 0.0069, P = 0.0191, respectively), and low AI (P = 0.0134, P = 0.0407, respectively) were significantly related to worse disease-free survival. Multivariate analysis showed AI as an independent factor predicting for disease-free survival (P = 0.0455)., Conclusions: The p53 mutations, expression of bax, and levels of spontaneous apoptosis have prognostic value in maxillary sinus SCC; AI especially is an independent factor for disease-free survival. A high level of spontaneous apoptosis induced by overexpression of bax may increase sensitivity of radiochemotherapy resulting in good prognosis, while p53 mutation may lead to resistance against radiochemotherapy, resulting in poor prognosis., (Copyright 2002 American Cancer Society.)

Chronic obstructive pulmonary disease (COPD) is characterized by periodic exacerbations, some of which are caused by nontypeable Haemophilus influenzae (NTHI). P6 is an outer membrane lipoprotein that is highly conserved among strains of NTHI. We hypothesized that lymphocytes from patients with COPD who have exacerbations due to NTHI have a decreased ability to recognize P6. The in vitro lymphocyte proliferative response to P6 in 36 patients with COPD and 12 healthy control subjects was studied. Ten patients who had exacerbations due to NTHI in the previous 12 months showed statistically significant lower proliferation to P6 (stimulation index, log transformed mean +/- standard error 0.82 +/- 0.17) compared with 26 patients who had no exacerbations due to NTHI in the previous 12 months (1.42 +/- 0.13) and to 12 healthy control subjects (1.61 +/- 0.16). These three groups had no significant difference in the lymphocyte proliferative response to tetanus toxoid. There was no difference in serum antibody levels to P6 in the two groups with COPD. These results indicate that decreased proliferation of T cells to P6 is associated with exacerbations of COPD and suggest that the ability of T cells to recognize P6 is associated with relative protection from exacerbations due to NTHI.

Children (n = 115; age range 1-9 years) with a diagnosis of acute otitis media (AOM) were eligible for this study and were enrolled within 24 h of the onset of symptoms/signs. A nasopharyngeal culture was obtained at the initial visit. Children were treated with a single oral antibiotic for 7 days. Changes in symptoms/signs and tympanic membrane features assessed by a scoring system were monitored for 1 month and related to the nasopharyngeal pathogen recovered on Day 1. More than 80% of children had no symptoms/signs by Day 3, and 60% of cases had normal tympanic membranes by Day 29. Children without any symptoms/signs on Day 3 had significantly lower symptom/sign scores on Day 1 (p = 0.005). Seventy-nine percent of cases carried middle ear pathogens in the nasopharynx at diagnosis: Streptococcus pneumoniae (48%); Haemophilus influenzae (24%); and Moraxella catarrhalis (16%). Children with S. pneumoniae showed significantly higher tympanic membrane scores than children with no pathogen at Days 8, 15 and 29 (p < 0.01 for each comparison). Multivariate regression analysis revealed that lower tympanic membrane score on Day 1, the absence of S. pneumoniae in the nasopharynx and treatment with amoxillin were independent factors for rapid normalization of the tympanic membrane. These data suggest that the clinical course of AOM may be predicted, in part, at the time of diagnosis by means of careful evaluation of symptoms/signs and the tympanic membrane as well as knowledge of pathogens harbored in the nasopharynx.

Treatment outcomes of tonsillectomy were studied in 7 Japanese patients with psoriasis--3 males and 4 females aged 9 to 46 years (median: 23 years)--followed up 2 to 9 years after tonsillectomy. All skin lesions disappeared in 3 patients, 80% of those in 2, and no change in the remaining 2 during follow-up. Of 5 in whom skin lesions improved, 4 were females and had a history of tonsillitis making skin lesions worse. In quantitative immunohistologic analysis on tonsillar tissues by CD20 and anti-ssDNA antibodies, areas of T cell-nodules were significantly expanded, but those of the B-lymphoid follicles were smaller, and the number of apoptotic cells increased in tonsils from patients with psoriasis and PPP compared to those with recurrent tonsillitis. The area of T cell-nodules and the number of apoptosis cells were significantly larger in tonsils from 4 patients with complete recovery after tonsillectomy compared to the remaining 9 without complete recovery. This suggests that histologic evaluation may be helpful in estimating the effectiveness of tonsillectomy.

Lymphoproliferative diseases of the nasal cavity and paranasal sinuses occur frequently in Asian countries and are histologically categorized as monomorphic ordinary lymphoma and polymorphic reticulosis (PR) with apparent inflammatory cell infiltration. The large atypical cells in PR show natural-killer cell nature and frequently contain Epstein-Barr virus (EBV) DNA. Among the EBV genes involved in latent infection, those encoding EBV latent membrane proteins are frequently expressed in PR. Several cytotoxic T-lymphocyte (CTL) defined epitopes have been mapped to latent membrane proteins restricted with HLA-A2, -A11 or -A24 antigens. Thus, the HLA-A allele may affect the development of PR. To examine this possibility, HLA-A alleles of 25 patients with EBV(+) PR were determined with low-resolution polymerase chain reaction-based typing using HLA-A locus sequence-specific primer combinations. The frequency of HLA-A alleles including HLA-A2 and -A24 antigens in PR patients was lower than that in the normal Japanese population, but the difference was not significant. Since HLA-A2-restricted CTL responses are well delineated at the A2-subtype level, the A2-subtype of PR cases with HLA-A2 antigen was further determined by high-resolution genetic typing. The frequency of HLA-A*0201 in PR was significantly lower than in the normal population (p=0.0314). The HLA-A*0201-restricted CTL responses may thus function in vivo to suppress the development of overt lymphoma., (Copyright 2000 Wiley-Liss, Inc.)

Prognostic factors and treatment outcome of 74 patients with non-Hodgkin's lymphoma of Waldeyer's ring (male 34, female 16, median age 63 years) and cervical lymph nodes (male 17, female 7, median age 62 years) were analyzed retrospectively. The prognostic factors analyzed were: age, B-symptoms, serum LDH levels, and Ann Arbor stage. We also analysed the TNM classification (1987) of Waldeyer's ring lymphoma (previously referred as nasopharyngeal and oropharyngeal cancer) as a new prognostic factor. The 5-year-survival rate of the patients with Waldeyer's ring non-Hodgkin's lymphoma was 67%, compared with 34% for cervical lymph node lymphoma. In univariate analyses, unfavorable outcome was associated with age (age > 60 years), B-symptoms, elevated serum LDH level, and stage. Multivariate analysis showed that age and elevated serum LDH level were significant independent risk factors for death. The 5-year-survival rate according to TNM classification of Waldeyer's ring lymphoma was: T1 (4 cases) 75%, T2 (23 cases) 83%, T3 (10 cases) 56%, T4 (4 cases) 0%; according to N stage, N0 (15 cases) 91%, N1 (7 cases) 73%, N2 (11 cases) 55%, N3 (5 cases), M1 (3 cases) 0%, and according to TNM stage, stage I (one case) stage II (9 cases) 100%, stage III (12 cases) 80%, stage IV (19 cases) 34%. These results suggest that the N classification of Waldeyer's ring non-Hodgkin's lymphoma may be a new prognostic factor.

Nasal lymphoma with polymorphic reticulosis (PR) morphology is now categorized as T/natural killer (T/NK) cell lymphoma. In this study, immunophenotypes and genotypes of proliferating cells in 21 cases with PR were examined. The patients included 13 men and 8 women ranging in age from 20 to 74 (median 37) years. All patients presented with lesions in the upper respiratory tract, mostly in the nasal cavity. Histological specimens obtained from the primary lesions (19 cases) and metastatic cervical lymph nodes (2 cases) were used for analyses. Histologically, polymorphous proliferation was found in 20 cases, and these were thus diagnosed as PR. A monomorphous pattern was found in the remaining last case. Immunohistochemical analysis revealed that the proliferating cells were CD56 (123C3)+ and/or CD16 (2H7)+, TIA-1+ and frequently stained CD3 epsilon+. Tumor cells were frequently stained positively with monoclonal antibodies (mAbs) for T lymphocytes, but were negative for T-cell receptor (TCR) beta and delta chain expression. In situ hybridization analysis using an Epstein-Barr virus-encoded early RNA 1 (EBER-1) probe revealed positive signals in 13 of the 15 cases examined. Southern blotting analysis for clonality of the Epstein-Barr virus (EBV) genome in 12 positive cases confirmed the presence of monoclonal proliferation in 7 cases. The pattern of TCR gamma chain gene rearrangement was examined by PCR analysis of DNA from tumor tissues by the denaturing gradient gel electrophoresis method. The results demonstrated no clonal rearrangement in any of the 21 cases examined, including 7 cases with proven clonal proliferation of EBV-infected cells, indicating the absence of T-cell clones. Our findings strongly suggested that nasal T-cell lymphoma is in fact a NK cell lymphoma.

To clarify the cytokine profile of tonsils, cytokine mRNA was determined by reverse transcription-polymerase chain reaction (RT-PCR). The mRNA of interleukin-1 beta (IL 1 beta), IL-2, IL-4, IL-6, IL-8, interferon-gamma (IFN-gamma) and tumor necrosis factor-alpha (TNF-alpha) were expressed on the whole tonsillar tissue of all 5 subjects with recurrent tonsillitis, obstructive sleep apnea syndrome caused by tonsillar hypertrophy or tonsillar focal infection. These cytokine mRNAs were detected in tonsillar mononuclear cells freshly isolated from all or 14 of 15 subjects. The CD4, CD8 and CD19-positive cells were purified using immunomagnetic beads. The CD4-positive cells expressed mRNA of IL-1 beta, IL-2, IL-4, IL-8, IFN-gamma and TNF-alpha. The CD8-positive cells expressed mRNA of IFN-gamma and TNF-alpha. The CD19-positive cells expressed mRNA of IL-1 beta, IL-6, IL-8 and TNF-alpha. Quantitative measurement of PCR products in tonsillar mononuclear cells revealed that the IL-8 mRNA was expressed at a higher level than any other cytokine. A comparison among tonsillar diseases mentioned above revealed that the IL-1 beta and TNF-alpha mRNA were expressed at significantly higher levels in tonsillar mononuclear cells from patients with recurrent tonsillitis than in patients with obstructive sleep apnea syndrome caused by tonsillar hypertrophy. These data indicate that tonsils are active immunologic organs containing a wide variety of cytokines.

Using two-color flow cytometry, we measured intracellular expression of interleukin-1 alpha (IL-1 alpha), IL-2, IL-4, IL-6, IL-8, interferon gamma (IFN-gamma) and tumor necrosis factor-alpha (TNF-alpha) in tonsillar mononuclear cells freshly isolated and stimulated by phorbol myristate acetate (PMA) and ionomycin. In freshly isolated tonsillar mononuclear cells, IL-1 alpha was produced in 0.39% of CD3 cells, 0.48% of CD4 cells, 0.66% of CD19 and 11.2% of CD14 cells; TNF-alpha was found in 5.4% of CD14 cells. After 8-hour culture without any mitogens, IL-4, IL-8, TNF-alpha and IL-1 alpha were detected in 2.1%, 0.8%, 0.55%, and 0.42% of tonsillar mononuclear cells, respectively. Flow cytometric detection of intracellular cytokines in tonsillar mononuclear cells stimulated with PMA and ionomycin revealed that CD3 cells produced IL-1 alpha, IL-2, IL-4, IL-8, IFN-gamma and TNF-alpha, and CD19 cells produced IL-1 alpha, IL-6, IL-8 and TFN-alpha. In CD3 cells, the number of cells producing IL-2 and TNF-alpha were significantly higher than those expressing other cytokines; and the number of cells producing IFN-gamma and IL-8 were significantly higher than those expressing IL-4 and IL-1 alpha. In CD19 cells, the number of cells producing IL-6 and TNF-alpha were significantly higher than those of IL-8 and IL-1 alpha; and the number of cells producing IL-8 was significantly higher than that of IL-1 alpha. There was no difference in the number of CD3 and CD19 cells producing any cytokine between the adult recurrent tonsillitis group and adult obstructive sleep apnea syndrome group. However, the number of CD3 cells producing IL-2 or TNF-alpha and CD19 cells producing IL-1 alpha, IL-6 or TNF-alpha were significantly lower in children than that of adults (p < 0.05). These findings indicate that the cytokine production in tonsillar mononuclear cells is heterogenous according to the subset and activation and that flow cytometric analysis of intracellular cytokines is a useful means to investigate the pathophysiological role of cytokines in the tonsils.

To determine the systemic immune response to alpha-streptococci (Str. sanguis, Str. salivarius and Str. mitis) and beta streptococcus (Str. pyogenes T12) in patients with tonsil related pustulosis palmaris et plantaris (PPP), we measured serum antibody levels to whole cell body antigens of Streptococcus (Str.) sanguis, Str. salivarius, Str. mitis or Str. pyogenes by enzyme-linked immunosorbent assay (ELISA). The serum IgG antibody levels to alpha-streptococci, Str. sanguis and Str. salivarius were significantly higher in patients with PPP (n = 44) than in patients with recurrent tonsillitis (RT: n = 25) and in healthy adults (n = 17). Serum antibody IgG level to Str. pyogenes was not different among the 3 groups. The IgA antibody levels against any Streptococcus strain were not different among the 3 groups. The IgM antibody levels to Str. pyogenes were significantly higher in patients with RT than in patients with PPP. In western blot analysis, the serum IgG antibodies against 25-27 kDa protein from whole cell body of Str. sanguis, Str. salivarius and Str. mitis were found more frequently in patients with PPP than in healthy adults. However, the western blot profile in Str. pyogenes was not different between PPP and healthy adults. No significant difference was seen in the western blot profile of IgM or IgM antibodies to any streptococcal whole cell bodies. These data suggest that systemic hyper immune response to alpha-streptococci may be present in patients with tonsil-related PPP and the 25-27 kDa protein of the organism may be the target for this immunologic abnormality.

In vitro cytokine production by tonsillar mononuclear cells under culture conditions with or without lyophilized streptococcal antigens was measured by enzyme-linked immunosorbent assay (ELISA). Under culture conditions without any stimulus, a certain amount of interferon (IFN)-gamma, tumour necrosis factor (TNF)-alpha, and interleukin (IL)-6, was detected in the supernatant culture fluids from all 29 donors tested and just detectable levels of IL-1alpha and IL-2 were identified in 5 of 12 donors tested; but IL-4 and IL-5 were not detected in any supernatants from 21 donors tested. In 16 patients with pustulosis palmaris et plantaris (PPP), all three alpha-streptococcal antigens from Streptococcus, (S.) sanguis, S. salivarius and S. mitis induced production of IL-6. IFN-gamma and TNF-alpha by tonsillar mononuclear cells at a significant level. However, no significant induction of such cytokines by alpha-streptococcal stimulus was seen in 13 donors without PPP. No significant difference in cytokine production by tonsillar mononuclear cells was seen between the PPP and non-PPP subjects, when stimulated with S. pyogenes, Staphylococcus aureus Cowan I and pokeweed mitogen. In the experiments using tonsillar cell subsets, the cytokine inductions by alpha-streptococcal stimulus were found in monocyte-depleted cells as well as in purified T-cells. but not in B-cells. These data suggest that IL-6, IFN-gamma and TNF-alpha may play an important role in immunological reactions in the tonsils and that a hyper-immune response to some of alpha-streptococcal antigens through increased production of such cytokines by tonsillar mononuclear cells, to which T-cells mainly contribute, may play a key role in the pathogenesis of PPP.

Cellular immune responses to the P6 outer membrane protein of non-typeable Haemophilus influenzae (NTHi) were determined in vitro by measuring immunoglobulin (Ig) secreting cells and lymphocyte proliferation in adenoidal and tonsillar lymphocytes from 19 children. Preliminary tests showed that P6 did not stimulate naive cells such as cord blood lymphocytes, but did stimulate sensitized cells in adenoids and tonsils. Cellular proliferation was significantly higher in adenoidal lymphocytes than in tonsillar lymphocytes (median: quadratile of stimulation index = 3.7:2.3-5.5 vs. 1.2:1.0-2.1, p < 0.02). A comparison between children with or without otitis media revealed that proliferative responses to P6 of adenoidal lymphocytes from children with otitis media were significantly decreased (2.0:1.8-3.6 vs. 3.7:2.3-5.5, p < 0.04). P6-specific antibody secreting cells were identified in a total of 14 adenoids and the number of cells secreting IgA was decreased in the otitis media group compared to controls (median: quadratile/10(6) cells = 435:359-499 vs. 755:593-1870, p < 0.05). Cultivation with P6 stimulated IgA secretion in children without otitis media, while no response was seen in children with otitis media (median: quadratile/10(6) cells = 1323:915-2410 vs. 2240:1900-2830, p < 0.02). These preliminary data demonstrate that lymphocytes from adenoids and tonsils recognize P6 as a specific antigen and that the adenoid is the more reactive of the two organs. Impaired P6-specific cellular immune responses of adenoids in children with otitis media may explain the recurrent nature of otitis media due to NTHi in the otitis prone population.

We measured the levels of serum IgG antibodies to CD outer membrane protein of Moraxella catarrhalis, P6 outer membrane protein of non-typeable Haemophilus influenzae and capsular polysaccharides of Streptococcus pneumoniae in 168 children with otitis media with effusion (OME) who were followed prospectively, using ELISA. Serum IgG antibodies to CD, P6 and pneumococcal capsular polysaccharides were detected in all samples. The anti-pneumococcal polysaccharides antibody level was highest, followed by the anti-P6 antibody level and anti-CD antibody was lowest (median:interquartile ranges were 45.9:19.1-100 microg/ml, 15.6:9.70-23.2 microg/ml and 1.06:0.73-1.87 microg/ml, respectively). In children aged 0-6 years, there were positive correlations among the antibody levels (anti-CD vs anti-P6, r=0.325, p <0.001; anti-CD vs anti-polysaccharide, r=0.397, p <0.0001; anti-P6 vs anti-polysaccharide, r=0.175, p=0.057). However, no relationship was seen in children aged 7-15 years. Children were classified according to severity of OME during the 1-year follow-up. In children aged 0-6 years, the severity of OME correlated inversely with the levels of anti-CD antibody (r=-.23, p=0.012), of anti-P6 antibody (r=-0.292, p=0.0015), and of anti-pneumococcal polysaccharides antibody (r=-0.25, p=0.0064). However, no correlation was found between antibody levels and severity of OME in children aged 7-15 years. These data suggest that persistence and/or recurrence of OME may be due to an insufficient serum antibody response to middle ear pathogens in young children.

A clinical study was performed of 83 patients with hypopharyngeal cancer treated in the Sapporo Medical University Hospital from 1982 to 1995. Five-year cumulative survival rate was 34.9% in the whole group and 37.2% in the radical surgical and/or radiation treatment group. In the radiation treatment group, the patients with T1, T2, N0 or N1 stage disease, especially those whose original disease responded almost completely to 40Gy irradiation, showed high tumor control rates following full-dose irradiation with or without radical neck dissection. In the radical surgery group, the patients with T4 or N2 stage disease showed better prognosis than those in the radiation group. However, among the patients with N3 lymph node metastases, there were no long-term survivors in either the radiation or the surgery groups.

This report clarifies the prognostic factors for survival in localized non-Hodgkin's lymphoma (NHL) of the head and neck and defines optimal regimens for this disease. One hundred-seven untreated patients with Stage I or II NHL of the head and neck were treated with involved field radiation therapy for orbital, nasal, or paranasal lymphoma and extended field radiation for Waldeyer's ring or neck lymphoma. Radiation doses were 39-48 Gy. In the latter half of the study, adjuvant chemotherapy was administered. Of 107 patients, 95 achieved complete response (CR). Of the 12 patients that did not achieve CR, 9 had nasal T-cell lymphoma (NTL) of the lethal midline granuloma type (LMG-NTL). Only one patient who obtained CR relapsed in a previously irradiated area. Age, sex, stage, bulky mass, number of involved sites, LMG-NTL, histologic subtypes, radiation dose, and adriamycin dose were analyzed for prognostic significance for disease-specific survival in NHL by multivariate analysis. LMG-NTL was the most significant prognostic factor (P < 0.001). Patients with higher age also experienced a higher relative risk than patients of > or =60 years of age (P = 0.0063). Dose of adriamycin reached the borderline significance (P = 0.0600). Radiotherapy is excellent for obtaining local control of head and neck NHL. Randomized trials are required to determine the appropriate radiation field and dose in patients previously treated with chemotherapy. LMG-NTL and age were the significant prognostic factors for disease-specific survival.

We report 10 postlingually deafened adults in whom the electrophysical criteria for cochlear implant were fulfilled, except that they showed the following unfavorable middle ear lesions: otitis media with effusion, chronic perforative otitis media, cholesteatoma and previous radical ear operation. Staged operations for cochlear implant were performed in 8 cases, and 2 patients who had undergone radical ear operation had a single-stage operation. As a first step, one of the following was performed in each patient as surgically indicated: myringoplasty with or without mastoidectomy, mastoidectomy with reconstruction of the posterior wall of the external canal, mastoidectomy with the insertion of a ventilation tube, radical mastoidectomy or surgical cleansing of the radical cavity. From 6 months to 2.5 years after the first operation, the actual cochlear implant was performed in the second or third stage. There was no major complication as a result of electrode insertion into the cochlea and the results of speech perception in these cases were not different from those in patients with normal middle ears. In our experience, it was considered that the staged operations would enable successful cochlear implants in selected patients with pathological middle ear lesions even if they had previously been diagnosed as contraindicated for this procedure. In a case with radical ear cavity a single-stage operation could be performed when there was no cavity problem.

In this report, we present 3 cases with thyroid carcinoma with tracheal invasion. In all cases, the tracheal defects were simultaneously reconstructed using a sternocleidomastoid muscle-clavicle myoosseous flap. We were able to close the tracheostomas without trouble, and no stenotic lesions or late complications were noted. This myoosseous composite flap has proved itself to be a reliable material for laryngotracheal reconstruction in some cases with thyroid carcinoma.

The authors reviewed the clinical, phenotypic, genotypic, and Epstein-Barr virus (EBV)-findings of 18 patients with nasal T-cell lymphoma(NTL). The clinical features were characterized as prolonged fever, widespread dissemination into distant sites, and poor prognosis with median survival of only 6 months. EBV-encoded small nuclear early region(EBER) transcripts were identified in 16 of 18 patients. Monoclonal EBV genome, EBV-encoded nuclear antigen(EBNA)-1, and latent membrane protein(LMP)-1 were also detected in all EBER-positive cases tested. All EBV-positive NTL showed coexpression of natural killer(NK) cell phenotype CD56 and CD2. Of 9 EBV-positive NTL, seven cases expressed T-cell receptor (TCR)-delta chain with rearranged beta-, gamma- and/or delta- genes. These data suggest that some cases of EBV-positive NTL may be derived from the lineage of NK-like T-cells or gamma delta T-cells, and that EBV may play a role in the lymphomagenesis.

Prognostic factors and treatment outcome of 71 patients with non-Hodgkin's lymphoma of Waldeyer's ring were analyzed retrospectively. In univariate analyses, unfavorable prognosis was associated with primary disease in the base of the tongue, stage III-IV diseases, B-symptoms, high-grade histology, T-cell phenotype, elevated serum LDH levels, decreased peripheral blood lymphocyte counts, and negative response on delayed type hypersensitivity skin reactions. Multivariate analysis showed that stage III-IV and T-cell phenotype were significant independent risk factors for death. In stage I-II lymphomas, patients with unilateral large or bilateral cervical lymph node involvement had a poorer prognosis. In stage I-II lymphomas with intermediate or high-grade histology, patients who had received radiotherapy with MTCOP-P chemotherapy (pirarubicin, cyclophosphamide, vincristine, methotrexate with leucovorin rescue, peplomycin, and predonisolone) showed significantly better 5-year disease-free survival rate compared with patients treated with radiotherapy alone.

Nasal T-cell lymphoma of the LMG type (LMG-NTL) is characterized by progressive, unrelenting ulceration, and necrosis of the nasal cavity and midline facial tissues. The clinical behavior of this tumor in 16 patients is compared with that of a nasal lymphoma of non-LMG-NTL type (non-LMG-NTL) in 8 patients and a paranasal sinus lymphoma (PSL) in 6 patients. All patients had stage I or II disease. Fourteen of the 16 patients with LMG-NTL received chemotherapy before and/or after radiotherapy. Cause-specific 5-year survival rates for patients with LMG-NTL, non-LMG-NTL, and PSL were 22%, 75%, and 67% respectively. Seven patients with LMG-NTL, had complete response, although 3 recurred, whereas it was incomplete in 9 patients. The data indicates that it is desirable to deliver 50 Gy or more to achieve in-field control of LMG-NTL.

Extranodal non-Hodgkin's lymphoma in the head and neck, especially T-cell lymphoma of the lethal midline granuloma (LMG) type, has unique clinical and histologic features differentiating it from other lymphomas. The authors measured soluble intercellular adhesion molecule-1 (ICAM-1) in sera from 12 patients with T-cell lymphoma of the LMG type and from 52 patients with other head and neck non-Hodgkin's lymphomas, by double-determinant immunoassay. The expression of ICAM-1 in lymphoma tissue was examined in 26 patients by the avidin-biotin immunoperoxidase method. The serum ICAM-1 levels were significantly higher in T-cell lymphoma of the LMG type than in other head and neck lymphomas or in healthy adult controls. Elevated levels of serum ICAM-1 were associated with increased levels of serum lactate dehydrogenase, poor prognosis, and impaired T-cell-dependent immune functions, both in T-cell lymphoma of the LMG type and in other head and neck lymphomas. When we monitored serum ICAM-1 levels in individual patients, the level decreased in the complete remission interval compared to before treatment and went up again when the lymphoma relapsed. Although the staining intensities of ICAM-1 in lymphoma cells were not related to serum ICAM-1 levels, a markedly intense expression of ICAM-1 was found on the angiocentric or angiodestructive lymphoreticular infiltrate region in the tissues from T-cell lymphoma of the LMG type. A higher serum ICAM-1 level and its tissue expression in T-cell lymphoma of the LMG type may be one of the clues to understanding this particular lymphoma. The serum ICAM-1 level could be an efficient parameter for monitoring the clinical course of head and neck non-Hodgkin's lymphomas.

Background: The authors have previously demonstrated nasal T-cell lymphoma (NTL) associated with Epstein-Barr virus (EBV). The detailed clinical, phenotypic, and genotypic features and the role of EBV in lymphomagenesis remain to be clarified., Methods: The study group consisted of 18 patients with NTL. The phenotype was determined by immunoperoxidase staining with various monoclonal antibodies. Genotypic study was done using Southern blot hybridization. The presence of EBV-encoded small nuclear early region (EBER) RNA and EBV DNA were determined by in situ hybridization. The expression of EBV-encoded nuclear antigen (EBNA) and latent membrane protein (LMP1) were identified by immunohistologic methods. Clonotypic analysis of EBV genomes was performed by Southern blot hybridization with EBV termini fragment probe., Results: The clinical features of NTL were characterized as prolonged fever (16 patients), widespread dissemination into distant sites (13 patients), and poor prognosis with a median survival of only 6 months. EBER transcripts were identified in 16 of 18 patients. Monoclonal EBV genomes EBNA1 and LMP1 were also detected in all EBER-positive cases tested. All 18 patients expressed pan-T antigens such as MT1, CD45RO, and/or CD2. The rearrangements of T-cell receptor (TCR)-beta, -gamma, and/or -delta genes were shown in all 11 patients tested. The natural killer (NK) cell phenotype CD56 was expressed in all EBV-positive cases tested, and was not detected in EBV-negative cases. Seven EBV-positive cases expressed a TCR-delta chain with rearranged TCR-gamma or -delta genes whereas both EBV-negative cases corresponded to alpha beta T-cell lymphoma, which expressed a TCR-beta chain with a rearranged TCR-beta gene., Conclusions: These data suggest that EBV-positive NTL may be derived from the lineage of NK-like T-cells or gamma delta T-cells, and that EBV may play a role in lymphomagenesis. Therefore, we propose that NTL which has peculiar clinical and histologic features could be classified as a new lymphoma entity.

The cellular immune responses to nontypable Haemophilus influenzae in adenoids were determined in children by measuring lymphocyte blast transformation and antibody secretion in response to the P6 outer membrane protein. In the lymphocyte transformation assay, stimulation index of adenoid lymphocytes stimulated by P6 in otitis children (2.34 +/- 0.25) was significantly lower than that in non-otitis children (3.91 +/- 0.64, p <0.05). The number of IgM as well as IgA secreting cells after 8 days' culture with P6 were significantly smaller in otitis children (IgM: 7,534 +/- 2,843/10(6) lymphocyte; IgA: 1,573 +/- 620/10(6) lymphocyte) than those in non-otitis children (IgM: 12,994 +/- 2,533, p <0.05; IgA: 2,828 +/- 528, p <0.05). These data suggest that P6 protein is a target for the cellular immune response of the adenoid, and failure of adenoid lymphocytes to recognize it as a specific immunogen may be one of the causes of recurrent otitis media.

We examined the expressions of alpha-subunits of very late antigen (VLA)-4 (alpha4) and VLA-5 (alpha5) on tonsillar lymphocytes and the interaction between these integrin receptors and their ligand, fibronectin (FN). Immunohistological and flow cytometric analyses showed that alpha4 and alpha5 were expressed in the lymphoid follicle and were positive on about 10% each of T cells and on 55% and 35% of B cells. When tonsillar B cells were separated by a discontinuous Percoll gradient, the number of alpha4- and 5-positive cells decreased as the cell density went down, while the number of activated cells went up. After in vitro activation of tonsillar B cells by Staphylococcus aureus Cowan I strain (SAC), the expression of alpha5 and the adhesiveness to FN decreased. The increased proliferation of B cells was observed when tonsillar B cells were cultured with immobilized FN. The expressions of alpha-subunits of VLA-4 and VLA-5 on tonsillar T and B lymphocytes increased with age. These results suggest that: i) B cell activation may cause decreased expressions of VLA-4 and -5, which gives a costimulatory effect on B cell activation itself again in cooperation with FN, ii) Increased expressions of VLA-4 and -5 on tonsillar lymphocytes with age may be related to regional immune response of the palatine tonsils.

The antibodies to LTA of Streptococcus pyogenes (S. pyogenes) and of Staphylococcus aureus (S. aureus) in sera from 110 children with or without recurrent tonsillitis were assayed by enzyme-linked immunosorbent assay (ELISA). The antibody of IgA class was not detected in any sera. There was a significant positive correlation between IgG and IgM antibody levels to LTA of S. pyogenes and those to LTA of S. aureus. There was no significant correlation between anti-LTA IgM antibody and age or episodes of tonsillitis. However, serum anti-LTA IgG levels in younger patients ( < or = 7 y.o.) were significantly lower than those in their elders (>7 y.o.) (anti-S. pyogenes; p = 0.0007, anti-S. aureus; p = 0.007). Serum anti-LTA IgG antibody levels correlated inversely with episodes of tonsillitis in younger patients ( < 7 y.o.) (S. pyogenes: r = -0.362, p = 0.0082, S. aureus: r = -0.276, p = 0.047). These findings suggest that systemic hyposensitivity to LTA of S. pyogenes and S. aureus may be a possible cause of habitualness of tonsillitis in young children.

To clarify the cellular immune response to alpha-streptococci in tonsillar lymphocytes from patients with pustulosis palmaris et plantaris (PPP) we have investigated cytokine production of tonsillar lymphocytes from patients with PPP recurrent tonsillitis (RT). Interleukin (IL)-1 alpha, IL-2, IL-4, IL-5, IL-6, interferon (IFN)-gamma and tumor necrotizing factor (TNF)-alpha were measured. The stimulation index of IFN-gamma production was significantly higher in tonsillar lymphocytes from PPP than that from RT when cultured with alpha-streptococci (stimulation index: the ratio of cytokine level produced by tonsillar lymphocytes with bacterial antigens to that without antigens). The same results were seen in the case of TNF-alpha, but IL-4 and IL-5 were not detected in any supernatants. These data suggest that increased production of IFN-gamma and TNF-alpha in tonsillar lymphocytes may play a key role in this kind of immunologic abnormalities.

Expression of various cytokines in the tonsillar compartments, and in vitro cytokine production and cytokine mRNA expression by tonsillar lymphocytes (TL), were investigated. In immunohistological analysis, a wide variety of cytokines such as interleukin (IL)-1 beta, IL-2, IL-4, IL-5, IL-10 was expressed intensively in the lymphoepithelial symbiosis (LES). A certain number of cells producing IL-6, interferon (IFN)-gamma, and tumor necrosis factor (TNF)-alpha was distributed not only in the LES but also in the extrafollicular area. A significant amount of IL-6, IFN-gamma, and TNF-alpha was detected by enzyme-linked immunosorbent assay in the culture supernatants of TL cultured with streptococcal antigen, even without any mitogens. Furthermore, mRNA of IL-6, IFN-gamma, and TNF-alpha was determined by reverse transcription polymerase chain reaction on the TL freshly isolated and with streptococcal stimulated antigen, whereas, production of IL-1 alpha, IL-4, and IL-5, and expression of their mRNA were not found on TL with or without streptococcal stimulation. These data indicate that tonsils are highly active immunological organs containing a wide variety of cytokine-producing cells, and suggest that IFN-gamma, TNF-alpha, and IL-6 may play an important role on the immune response to bacteria in the tonsil.

To determine whether EBV-specific cellular immunity persists and controls EBV in the palatine tonsil, we compared EBV-specific cellular immune responses in the palatine tonsil and peripheral blood. In 32 EBV-seropositive donors, we performed in vitro growth inhibition assays of EBV-induced B-lymphocyte transformation (regression assay). The results showed that EBV-specific cytotoxic T lymphocytes (CTL) precursor frequency was higher in the palatine tonsil than in the peripheral blood. After a first stimulation with autologous lymphoblastoid cell line (LCL), the EBV-specific cytotoxicity of reactivated CTL was higher in the palatine tonsil than in the peripheral blood. Furthermore EBV-specific CTL lines established from the palatine tonsil had significantly higher EBV-specific cytotoxicity in comparison with those from the peripheral blood. Both the EBV-specific CTLs from tonsils and peripheral blood were restricted by HLA class-I determinants. These data indicate that the palatine tonsil, which is frequently exposed to EBV, plays an important role in controlling EBV infection in the oropharynx.

Eighteen patients with a 22-channel cochlear implant were evaluated both pre- and postoperatively with vestibular function tests; caloric test and body balance test using stabilometry. Preoperative promontory stimulation results and postoperative psychophysical factors of the cochlear implant showed no correlation with the preoperative vestibular function test results. The results of speech perception tests were not affected by the degree of residual vestibular function in any of the patients. However, for patients with deafness for a period of 10 to 20 years, the degree of the residual vestibular function estimated by the caloric test showed a correlation with the results of speech perception tests, significantly in the consonant recognition score. The results, not of length or the Rombberg ratio, but rather of the area of body sway, showed a correlation with the results of speech perception tests; consonant recognition, in patients with a short duration of deafness. Therefore preoperative vestibular function results may be useful as a predictive factor in postoperative speech perception; consonant recognition, for patients with a short duration of deafness. Reduced results of caloric testing after surgery were found in one of 11 patients (9%) with residual vestibular function. The occurrence of vertigo immediately after surgery was observed in 5 of 18 patients (28%), but this symptom was transient. No change in stabilometry results, pre-versus post-operatively, was observed. Thus, vestibular damage after surgery was considered to be minimal.

Superoxide anion (O2-) production by peripheral blood polymorphonuclear leukocytes (PMNs) stimulated with phorbol myristate acetate (PMA) was measured by the cytochrome C method in 57 patients with recurrent tonsillitis. There was no significant correlation between superoxide production and episodes of acute tonsillitis, serum C-reactive protein levels, or peripheral white blood cell count. However, the superoxide production by PMNs was inversely correlated with serum anti-streptolysin-O titers (r = -.38, p = .008). Furthermore, the mean +/- SD superoxide levels produced by PMNs from patients with high serum ASO titers (> 71 U/ml), 69.58 +/- 30.56 nM/10(6) cells, was significantly lower than that of patients with low serum ASO titers (< 71 U/ml), 89.83 +/- 38.90 nM/10(6) cells (p = 0.037), and that of healthy adult controls, 102.27 +/- 44.67 nM/10(6) cells (p = 0.012). In addition, the effect of Streptococcus pyogenes on superoxide production by PMNs was studied in vitro. Superoxide production by PMNs preincubated with 600 micrograms/ml culture supernatant of Streptococcus pyogenes T4 (not detected) and T12 (34.82 +/- 31.40 nM/10(6) cells) was significantly lower than that of PMNs preincubated with culture medium (136.09 +/- 70.41 nM/10(6) cells; p < 0.05, each). Inhibition of superoxide production by PMNs by preincubation with culture supernatant of Streptococcus was increased by the protein in the culture supernatant in a concentration-dependent manner. These findings suggest that frequent and/or persistent streptococcal infections may reduce the superoxide production by PMNs, leading to recurrent episodes of tonsillitis.

The relationship between nasopharyngeal colonization with nontypeable H. influenzae and recurrent otitis media was assessed in 157 children followed prospectively from birth through 12 months of age. Forty-nine (31%) became colonized. Nasopharyngeal secretory IgA (sIgA) reactive with the P6 outer membrane protein was detected in all colonized children. Reduction or elimination of the organism was associated with a better mucosal immune response (560 +/- 864 units/ng/mL of sIgA) than was persistence in the nasopharynx (121 +/- 81; P = .04). Forty colonized children (82%) and 61 noncolonized children (56%) developed otitis media (P = .004); colonized children were four times more likely to be classified as otitis prone (P = .003). The frequency of otitis media episodes was directly related to the frequency of colonization (r = .42, P < .01). These results demonstrate a strong relationship between nasopharyngeal colonization patterns and otitis media. The mucosal immune response may be important in elimination of potential pathogens from the respiratory tract.

Nasopharyngeal colonization with Moraxella catarrhalis was evaluated in a large cohort of infants followed prospectively from birth to 2 years of age; 120 children were examined at 13 routine visits. Of these, 66% became colonized with M. catarrhalis by 1 year and 77.5% by 2 years. Nasopharyngeal colonization with M. catarrhalis increased from 27.0% during healthy visits to 62.7% during visits due to otitis media (P < .001). Otitis-prone children were colonized at 44.4% of all visits compared with 16.7% for children who did not have otitis media (P < .001). DNA from 112 strains of M. catarrhalis from 34 children were evaluated; 106 were successfully digested with restriction enzymes and demonstrated a great degree of heterogeneity. Children tended to acquire and eliminate a number of different strains. Intrafamilial spread of the same strain of M. catarrhalis was frequent. These data suggest that nasopharyngeal colonization with M. catarrhalis is common throughout infancy. A high rate of colonization is associated with an increased risk of otitis media.

Epstein-Barr virus (EBV)-specific cytotoxic activities of tonsillar lymphocytes (TL) and peripheral blood lymphocytes (PBL) obtained from the 26 same donors were investigated. The minimum cell concentration of TL required for regression of EBV-transformation, 3.1 +/- 2.1 x 10(4)/ml, was significantly lower (p < .01) than that of PBL, 4.3 +/- 2.6 x 10(4)/ml, indicating that the EBV-specific cell-mediated cytotoxicity of TL was significantly higher than that of PBL. The cytotoxic activity against an autologous EBV-positive B-lymphoblastoid cell line (LCL) of the tonsillar T-lymphocytes, 10.8 +/- 3.7%, was significantly higher than that of the peripheral blood T-lymphocytes, 5.9 +/- 1.6%, after co-cultivation with an autologous-LCL (p < 0.05). The EBV-specific cytotoxic T-lymphocytes (CTL) lines, which contained > 95% CD8-positive cells, established from TL, had significantly higher cytotoxic activities against autologous-LCL (73.1 +/- 16.5%) than did the CTL lines established from PBL (59.0 +/- 16.4%, p < .01). The number of tonsillar T-lymphocytes which showed regression against in vitro EBV-transformation was significantly higher in patients with tonsillar focal infection, 2276 +/- 901 cells/well, than in patients with simple tonsillar hypertrophy, 991 +/- 350 cells/well, (p < .05), suggesting that the regression activity of tonsillar T-lymphocytes was significantly lower in patients with tonsillar focal infection than in patients with simple tonsillar hypertrophy. These data show that TL have higher EBV-specific cytotoxicities than PBL, suggesting that the tonsils may play an important role in defense against EBV.

Sixty-eight children fed human milk were followed prospectively from birth to 12 months of age to assess the effect of milk antibody on nasopharyngeal colonization. Human milk secretory IgA antibody to P6, a highly conserved outer membrane protein of nontypeable Haemophilus influenzae, was measured with the use of an enzyme-linked immunosorbent assay. Nasopharyngeal colonization with nontypeable H. influenzae and the occurrence of otitis media were determined. Nasopharyngeal colonization was found in 22 children (32%), and 39 children (57%) had otitis media. Frequency of isolation of nontypeable H. influenzae was directly related to episodes of otitis media (r = 0.35; p = 0.001). The level of human milk anti-P6 secretory IgA antibody was inversely related to frequency of isolation of the organism (r = -0.27; p = 0.026). The average antibody level, expressed as nanograms per 0.1 mg total secretory IgA, in human milk fed to children with no colonization of nontypeable H. influenzae was significantly higher than in milk fed to children in whom colonization occurred on multiple occasions (156 +/- 120 vs 69 +/- 50; p = 0.013). Prevention of colonization was most evident during breast-feeding. These data suggest that the protective effects of human milk against otitis media may be due in part to inhibition of nasopharyngeal colonization with nontypeable H. influenzae by specific secretory IgA antibody.

Adult T-cell leukaemia-lymphoma (ATLL) is a new type of T-cell malignancy which has an adult onset, a rapidly progressive terminal course and a clustering at patients' birthplaces in southeast Japan and the Caribbean. We report the clinical features of three cases of ATLL in Waldeyer's ring, which has been rarely reported in the literature. The patients complained of throat pain and lump in the neck. They received combination chemotherapy, but all died within a year of first noticing the symptoms.

So-called lethal midline granuloma is of great clinical and theoretical interest. The etiology of lethal midline granuloma is unknown and the pathogenesis is variable, with debate as to precise classification and natural history. In this study, we reported genotypic and immunopathological features in 3 cases of lethal midline granuloma. The histopathological diagnosis of their biopsy specimens was initially polymorphic reticulosis/midline malignant reticulosis. Immunohistologic study of the specimens revealed that immature or atypical cells had phenotypes of T-cells, CD2, CD3, CD4 (Case 1), CD4 (Case 2), and CD2, CD3 (Case 3). Those cells were also found to be positive for HLA-DR, which indicated that they were activated T-cells. Immunohistology in T-cells, however, was not able to give a similar clue to clonarity as it was possible within B-cell neoplasms by immunophenotyping the light chains. With the establishment of cDNA probes for the T-cell receptor genes it was possible to analyze neoplasms of lymphocyte origin for lineage and clonality. The Southern blot analysis of 3 cases showed rearrangement of TCR gene, TCR beta and TCR gamma chain (Cases 1 and 2) and TCR beta and TCR delta chain (Case 3), whereas none of them showed rearrangement of immunoglobulin heavy chain. These findings represented conclusive evidence for a monoclonal T-cell proliferation within lethal midline granuloma. On the ground of immunohistological and genotypic studies, lethal midline granuloma histologically diagnosed as polymorphic reticulosis/midline malignant reticulosis are proven to be a T-cell lymphoproliferative disorder.

The distribution of lymphoid cells in the mantle zone, germinal center, interfollicular area, and subepithelial area of the tonsil was evaluated quantitatively by image analysis in 66 subjects aged 3 to 66 years. The number of Ig-positive cells in the tonsil decreased with advancing years in all compartments. This inverse correlation to age was statistically significant for IgD-, IgM-, and IgG-positive cells. For T-cells, overall change of each T-cell subset with age was smaller than those of Ig-positive cells. An age-related marked decline was seen for CD4-positive cells only in the subepithelial area and for CD8-positive cells only in the interfollicular area. Ki-67-positive cells, cells undergoing active division, were mainly found in the germinal centers and also diminished with advancing years. Patients with frequent episodes of tonsillitis demonstrated a significant increase of IgD-positive cells and IgG-positive cells in interfollicular and subepithelial compartments and a decrease of CD4-positive T-cells in the germinal centers and subepithelial areas. These results suggest that the tonsillar involution with age is immunologically associated in all compartments with the decrease of Ig-positive cells and Ki-67-positive activated cells resulting in a relative increase of T-cell subsets. The method of image analysis provides a novel and unique approach for quantitative immunohistological study of the tonsil.

Distribution of various T-cell subsets in the palatine tonsil was investigated by two-color flow cytometry and double immunoenzymatic stain. Tonsillar lymphocytes contained many (about 20%) helper T (CD4+ Leu8-) cells and few (only 1%) suppressor T (CD8+ CD11b+) cells. In interfollicular area, each T-cell subset, i.e., helper, helper-inducer (CD4+ CD29+), suppressor-inducer (CD4+ CD45RA+), cytotoxic (CD8+ CD11b-), and suppressor, was identified by double immunoenzymatic labeling technique using alkaline phosphatase and peroxidase. On the other hand, only two T-cell subsets, helper, and cytotoxic T-cell subpopulations, were recognized in germial center. These results indicate that double immunoenzymatic stain as well as two-color flow cytometry gives us useful informations in terms of tonsillar T-cell subsets.

The distribution of Epstein-Barr virus receptor (EBVR) among the different B-cell subpopulations was analyzed by flow cytometry, using simultaneous two-color immunofluorescence of EBVR and cell-surface markers. The expression of EBVR was established by the binding of fluorescein isothiocyanate (FITC)-labeled EBV to the cells, while surface markers were stained by phycoerythrin (PE)-indirect immunofluorescence, using monoclonal antibodies. All cells of both the resting B-cell subpopulation defined by L30 and the B-cell subpopulations expressing surface IgM, IgD, IgA, and IgG were EBVR-positive. In contrast, EBVR was absent from about 10% cells of the activated B-cell subpopulation recognized by OKT9, as well as from about 10% cells of the highly differentiated B-cell subpopulation which reacted with OKT10. These results suggest that the expression of EBVR on the B-cell lineage varies with the maturation stage and with the state of activation. This postulation was further supported by analyses based on in vitro B-cell activation.

Using immunoperoxidase technique and various monoclonal antibodies, B1, B2, OKT9, OKT10 and Leu-7, we investigated B cell differentiation in the tonsillar follicle. Mantle zone was stained with B1 intensely and B2 faintly. In contrast, germinal center was stained with B1, OKT10 and OKT9, and more intensely with B2. In the intermediate part of the germinal center some large cells were stained with OKT10. With OKT9 antibody, most of cells in the lymphoepithelial symbiosis and some large dendritic cells considered to be tangible body macrophages or dendritic reticulum cells in the germinal center were intensely stained. Leu-7 positive cells were localized mainly in the intermediate part of the germinal center. Stages of B cell differentiation in the tonsillar follicle were discussed, considering these results.

The specific antibody response of the adenoids to a respiratory antigen was investigated, both in vitro and in vivo. The in vitro immunoglobulin (Ig) production of adenoidal and tonsillar lymphocytes in the same patient in 18 cases was measured by an enzyme-linked immunosorbent assay. Adenoidal lymphocytes produced more IgM than IgG or IgA under culture conditions without any mitogens, whereas IgG was the major Ig produced by tonsillar lymphocytes. The same results were obtained when cultured with pokeweed mitogen. Under culture conditions with Dermatophagoides farinae (mite) antigen, adenoidal lymphocytes produced only specific IgM class antibody, and at a level significantly greater than tonsillar lymphocytes. Through an in vivo study, we established experimental adenoidal tissue in the guinea pig by long-term exposure to an ovalbumin aerosol. Infiltration of cells producing the specific antibody against an ovalbumin was demonstrated in the epipharyngeal mucosa using immunofluorescent staining. Our results confirm that the adenoids play a role in specific immune responses to respiratory antigens.

We investigated host-immune defenses against head and neck cancer cells by using various monoclonal antibodies with an immunoperoxidase technique to define lymphocyte subsets and natural killer (NK) cells. By so doing, we were able to identify lymphocyte subsets and NK cells in various head and neck cancers. We found that the majority of these cells infiltrate in or around nests of cancer cells and are stained with Leu-1 antibody. They include both Leu-2a and Leu-3a positive cells, which show equally intense levels of infiltration. Leu-7 positive cells were only scattered in the peripheral portion of the cancer nests in some cases. We also found a tendency for T-cells to infiltrate more intensely in poorly differentiated squamous cell carcinomas (SCC) than in moderately or well-differentiated SCC. Similarly, T-cells were more prevalent in maxillary carcinomas than in laryngeal carcinomas. These findings suggest the presence of a host-immune defense mechanism against cancer cells in patients with head and neck cancers.

Employing various monoclonal antibodies and immunoperoxidase technique, we studied the distribution of the activated lymphocytes in human tonsillar tissue. Both the activated T-cells positive for IL2-receptor (IL2-R) and the activated B-cells defined by L29 were seen in the interfollicular area. Vast majority of the lymphocytes in the germinal center was stained with the L29, the anti-transferrin-receptor antibody, and the Ki-67 antibody which reacts with cycling cells in G1, S, or G2 + M phase. On the other hand, scarcely any cells in the mantle zone were stained with those. The Ki-67 positive cells in the germinal center were identified as two types of staining pattern, i.e., with strong nucleolar staining found in the dark zone and with nuclear staining found in the light zone. Small number of IL2-R positive cells was found in the mantle zone. The cell number per unit area of the activated lymphocytes in the interfollicular area as well as in the germinal center was decreased as the increment of patients' age. From these results, the immunological activation system of T- and B-cells in the tonsils was discussed.

Employing a series of monoclonal antibodies directed against T cell subsets and the ABC (avidin-biotin complex) method as the immunoperoxidase technique, we studied the distribution of T cell subsets in frozen tissue sections of tonsils in 8 cases with various tonsillar lesions. Anti-Leu 1 and anti-Leu 4 antibodies, which react with all peripheral T cells, stained a majority of interfollicular cells and mononuclear cells in the lymphoepithelial symbiosis (LES). Approximately 80% of Leu 1+ and Leu 4+ cells were reactive with anti-Leu 3a antibody, which defines the helper/inducer T cell subset. Small lymphocytes in the mantle zone of the tonsillar follicle had IgD and IgM on their surface, whereas the cells in the germinal center showed membrane and/or cytoplasmic staining with anti-IgM serum. In addition, Leu 3a+ cells were found in the germinal center, especially on the capsular side showing a linear distribution. On the other hand, anti-Leu 2a antibody, which defines the suppressor/cytotoxic T cell subset, stained only a minority of lymphocytes in the interfollicular area and in the LES. Leu 2a+ cells were rarely found in the germinal centers. Anti-HLA-DR-positive cells were found with two types of staining pattern, i.e., with membrane staining found in the small lymphocytes in the mantle zone and the germinal center and with membrane and/or cytoplasmic staining found in large cells with dendritic processes in the germinal center, in the LES, and in the interfollicular areas. These findings demonstrate the presence of intimate cellular interactions in T cells, B cells, and macrophages in the tonsil, and indicate that immunohistological study of the tonsil may provide new insights into the pathogenesis of tonsillar lesions.