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Biomedical subjects

Y Harabuchi

Publications and source records attributed to Y Harabuchi.

At least 55 records · Page 3Linked to original sources

Human milk secretory IgA antibody to nontypeable Haemophilus influenzae: possible protective effects against nasopharyngeal colonization.

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.

Acute Disease↗

Epidemiology of Moraxella catarrhalis in children during the first 2 years of life: relationship to otitis media.

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.

Child, Preschool↗

Nasopharyngeal colonization with nontypeable Haemophilus influenzae and recurrent otitis media. Tonawanda/Williamsville Pediatrics.

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.

Age Factors↗

Clonality, expression and methylation patterns of the Epstein-Barr virus genomes in lethal midline granulomas classified as peripheral angiocentric T cell lymphomas.

We analysed the terminal repeats of Epstein-Barr virus (EBV) in DNAs isolated from six lethal midline granuloma (LMG) biopsies. A single fused terminal fragment could be detected in each case, indicating that these angiocentric peripheral T cell lymphomas represent clonal proliferations of cells infected with EBV on a single occasion. Using reverse transcriptase-PCR, we detected EBV nuclear antigen (EBNA) 1 and latent membrane protein (LMP) 1, but not EBNA 2 messages in LMG biopsy RNAs. The splicing pattern of the EBNA 1 message was consistent with the usage of a promoter localized in the BamHI F fragment (F promoter). The BamHI W fragment repeats and LMP-coding sequences were highly methylated in all cases. In contrast, the LMP regulatory sequences were found to be hypomethylated or partially methylated, as in LMP-expressing nasopharyngeal carcinomas.

Antigens, Viral↗

[Superoxide production by neutrophils in patients with recurrent tonsillitis].

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.

Adolescent↗

[Analysis of Epstein-Barr virus-specific cellular cytotoxicity in tonsillar lymphocytes].

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.

Adolescent↗

Adult T-cell leukaemia-lymphoma in Waldeyer's ring: a report of three cases.

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.

Adult↗

The prognostic value of Ki-67 antigen in non-Hodgkin lymphoma of Waldeyer ring and the nasal cavity.

BACKGROUND: A monoclonal antibody, Ki-67, recognizes an antigen expressed in all phases of the cell cycle, except G0, and can be used as a simple histologic marker of cell proliferation. To assess the prognostic value of the growth fraction in non-Hodgkin lymphoma of Waldeyer ring (W-NHL) and the nasal cavity (N-NHL), the authors applied Ki-67 immunostaining combined with image analysis on such lymphomas. METHODS: The authors studied 29 patients (18 with W-NHL and 11 with N-NHL), applying Ki-67 to frozen sections. The number of Ki-67-positive cells in a unit area (0.044 mm2), as an indicator of proliferative activity, and the mean area per Ki-67-positive cell (microns2), as an indicator of DNA content, were measured by the image processing system. RESULTS: High-grade lymphomas showed a significantly larger number of Ki-67-positive cells than intermediate-grade lymphomas (102.5 +/- 21.6 in high-grade and 46.8 +/- 8.92 in intermediate-grade lymphomas, P = 0.03), even when analyzed separately by immunophenotypes. A large mean area per Ki-67-positive cell was associated significantly with a T-cell phenotype (36.3 +/- 7.69 microns2 in T-cell lymphomas and 19.4 +/- 2.33 microns2 in B-cell lymphomas, P = 0.034) and an unfavorable clinical outcome. High proliferative activity, defined as nuclear Ki-67 expression in 2000 or more B-cell lymphoma cells and 1000 or more T-cell lymphoma cells in a 1-mm2 area, was found to be a strong predictor of poor survival among these patients (P = 0.048 and P = 0.009, respectively). CONCLUSIONS: Ki-67 immunostaining, combined with image analysis, is a novel method for determining a tumor proliferative index that provides useful clinical data regarding head and neck lymphomas.

Adolescent↗

Lethal midline granuloma (peripheral T-cell lymphoma) after lymphomatoid papulosis.

A Japanese woman with an 8-year history of lymphomatoid papulosis (LP) had lethal midline granuloma (LMG) develop at the age of 51 years. There were histologic similarities between LP and LMG seen in this patient. Surface phenotypic studies on nasal and cutaneous lesions demonstrated a population of T-cells expressing CD2, CD4, CD25, CD30, and histocompatibility antigen-DR (HLA-DR). Genotypic analyses of nasal and skin biopsy specimens disclosed a clonal rearrangement of the beta T-cell receptor gene with the same rearrangement pattern. These data indicate that this patient had LMG characterized by clonal peripheral T-cell lymphoma, which probably resulted from progression of the LP.

Adult↗

Distribution of lymphoid cells in tonsillar compartments in relation to infection and age. A quantitative study using image analysis.

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.

Adolescent↗

[Genotypic analysis of lethal midline granuloma].

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.

Adult↗

Epstein-Barr virus in nasal T-cell lymphomas in patients with lethal midline granuloma.

Five cases of lethal midline granuloma were identified histologically and phenotypically as peripheral T-cell lymphomas. Epstein-Barr virus (EBV) DNA was detected in the nasal tumour biopsy specimens by Southern blotting and in-vitro hybridisation with simultaneous detection of EBV-determined nuclear antigen (EBNA) and T-cell surface markers by two-colour immunofluorescence. Further immunofluorescence and northern blotting revealed that EBNA2 gene and also latent membrane protein gene were expressed in the nasal tumour cells. The patients had high titres of antibodies to EBV. These findings suggest that lethal midline granuloma is causally associated with EBV.

Adult↗

[Double immunoenzymatic and two-color flow cytometric analyses of tonsillar T-cell subsets].

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.

Flow Cytometry↗

[Immunohistological identification of the activated lymphocytes in human palatine tonsil].

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.

Adolescent↗

Soluble immune complexes and squamous cell carcinoma-related antigens in patients with head and neck cancer.

Sera from 85 patients with head and neck cancer including laryngeal cancer, hypopharyngeal cancer, nasopharyngeal cancer, and maxillary cancer were assayed for immune complexes (IC) by solid-phase anti-C3 enzyme immunoassay and for squamous cell carcinoma-related (SCC) antigen by radioimmunoassay. The positive rates of IC and SCC antigen in head and neck cancer patients were 29.4% and 34.1%, respectively. Their serum levels and positive rates were found to be elevated according to the degree of advancement of the disease stage, showing their good clinical correlations. With individual patients there was no significant relationship between IC and SCC antigen.

Adolescent↗

Immunosuppressive substance in the sera of head and neck cancer patients.

Serum levels of immunosuppressive substance (IS) and immunosuppressive acidic protein (IAP) were studied by single radial immunodiffusion in 108 patients with head and neck cancer. Mean values of serum IS and IAP in patients with head and neck cancers were as follows: laryngeal carcinoma, 955 +/- 447 micrograms/ml (IS), 604 +/- 324 micrograms/ml (IAP); maxillary carcinoma, 896 +/- 384 micrograms/ml (IS), 568 +/- 246 micrograms/ml (IAP); nasopharyngeal carcinoma, 871 +/- 313 micrograms/ml (IS), 673 +/- 395 micrograms/ml (IAP); hypopharyngeal carcinoma, 1034 +/- 495 micrograms/ml (IS), 669 +/- 335 micrograms/ml (IAP). Except for IAP in hypopharyngeal carcinoma, these values were significantly higher than those of controls (IS, P less than 0.01; IAP, P less than 0.05). The overall positive rates of IS and IAP in the patient group were 46% and 37%, respectively. In patients with laryngeal carcinoma the positive rates of serum IS and IAP were found to be elevated as the disease stage advanced. The IS showed a higher positive rate than IAP in the advanced disease stage. Serial studies revealed that all patients who had recurrent disease after the treatment exhibited marked elevation of serum IS levels; in addition, patients with recurrent cancer showed significantly higher IS levels (1507 +/- 464 micrograms/ml) compared to patients who had remained free of disease in the follow-up period (640 +/- 143 micrograms/ml) (P less than 0.001). These data strongly suggests that serum IS and IAP levels are useful parameters for monitoring the disease stage of head and neck cancer patients.

Adolescent↗

Flow cytometric analysis of Epstein-Barr virus receptor among the different B-cell subpopulations using simultaneous two-color immunofluorescence.

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.

B-Lymphocytes↗