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K Kuwano

Publications and source records attributed to K Kuwano.

At least 109 records · Page 6Linked to original sources

[Cross reactive antigens of Acholeplasma laidlawii and L-form of Staphylococcus aureus].

We demonstrated that the membrane of Acholeplasma laidlawii PG8 and L-form of Staphylococcus aureus, both of which induce cellular immunity in BALB/c mice, were antigenically related each other. Foodpad responses of the mice immunized with a mixture of either antigen and Freund's complete adjuvant showed clearly a cross reaction when challenged with the other antigen. Cross responses to incorporate 3H-thymidine to the spleen lymphocytes of the mice immunized with either antigen occurred in the presence of the other antigen. Furthermore, the purified T cells, but not B cells, of the spleen were activated in the presence of antigen-presenting cells. These antigens existing in the membrane fractions of both microorganisms were purified by Razin's method. Finally, these membrane components of A. laidlawii and L-form of S. aureus were subjected to gel electrophoresis and transferring to nitrocellulose membrane and used to stimulate the spleen lymphocytes of the mice immunized with A. laidlawii or of non-immunized mice. The fractions representing molecular weights of approximately 45 kD, 25 kD, and 13 kD of both microorganisms consistently stimulated the lymphocytes of the immunized mice but not those of non-immunized mice.

Acholeplasma laidlawii↗

[The CT diagnosis of emphysema and correlation with pathologic findings].

It is difficult to diagnose clinically silent or mild emphysema. The efficacy of CT scans for the diagnosis of emphysema has attracted attention and comparisons have been made between CT images and the pathological grade of emphysema in resected lungs. With a view to determine to what extent high resolution CT images are accurate concerning the diagnosis of mild emphysema, we conducted an extensive comparative study on CT scores of emphysema, based on high resolution CT, the pathology score derived from the cut surface of the lung identical in its plane with that of the CT and destructive index (DI) which is said to be instrumental in representing the degree of deterioration of alveolar walls in the same regions of the lungs. In this study, 42 patients who underwent thoracotomy and their lung specimens including a solitary nodule of considerable size have been employed. The CT scores and pathology scores depend on the picture grading system developed by Thurlbeck and coworkers for the basis of counting. The findings were such that with 1 mm collimation, the CT scores ranged between 12 and 57 with a mean +/- SD of 22.1 +/- 9.6 (n = 35) while with 5 mm collimation, the CT scores ranged between 7 and 46 with a mean +/- SD of 16.5 +/- 8.3 (n = 33). The pathology scores stood at 10 to 57 with a mean +/- SD of 23.2 +/- 9.8 (n = 42).(ABSTRACT TRUNCATED AT 250 WORDS)

Adult↗

[Evaluation of pulmonary emphysema with high resolution computed tomography and the effect of smoking].

Cigarette smoking is thought to play an important part in the development of pulmonary emphysema. The authors previously reported on the correlation of high resolution computed tomography (CT) with the pathology grade of emphysema and the destructive index of lung specimens from 42 patients undergoing thoracotomy for a solitary pulmonary nodule. High resolution CT was effective to identify the presence and grade of mild emphysema. The mean per cent predicted FEV1 was 97 per cent. There were 28 smokers and 14 nonsmokers. In this report, the correlation of smoking history with pathology score and high resolution CT was analyzed. There was significant correlation among these indices. These results further support the relationship between smoking and pulmonary emphysema. High resolution CT scan may help in the understanding of the role of cigarette smoking in the development of early pulmonary emphysema.

Adult↗

Cross-reactive protection against influenza A virus infections by an NS1-specific CTL clone.

An influenza A subtype cross-reactive CTL clone (A-11) was established following stimulation of A/PR/8 virus-immune spleen cells of Balb/C (H-2d) mice. This T cell clone lysed target cells infected with influenza viruses of the H1, H2, or H3 subtypes, and recognizes a conserved epitope on the NS1 protein. The clone is restricted by the H-2Ld allele. Adoptive transfer of A-11 significantly reduced virus titers in the lungs of mice infected with influenza A viruses of the H1, H2, or H3 subtypes. These results suggest that the conserved epitope on NS1 which is recognized by A-11 may be a useful component to consider for inclusion in experimental cross-reactive influenza vaccines.

Alleles↗

Enhancement of cytotoxicity of active macrophages by mycoplasma: role of mycoplasma-associated induction of tumor necrosis factor-alpha (TNF-alpha) in macrophages.

Tumor necrosis factor-alpha (TNF-alpha)-inducing activity of several mycoplasmas including Mycoplasma pneumoniae, a causative agent in human respiratory infectious diseases, was investigated. Purified peritoneal macrophages from BALB/c mice markedly enhanced their cytotoxic activity to Meth A cells, when cultured with either viable or non-viable mycoplasmas. The supernatants of the macrophage culture with mycoplasmas, M. pneumoniae and Acholeplasma laidlawii, showed the potent cytotoxic activity to TNF-alpha-sensitive L cells but not to TNF-alpha-insensitive L cells. Addition of anti-TNF-alpha antiserum inhibited completely the cytotoxic activity of these supernatants, indicating that a major part of the cytotoxic activity might be due to TNF-alpha. Various other mycoplasmas, either glucose- or arginine-utilizing species, as far as tested showed also the potent activity to produce TNF-alpha. These results strongly suggest the possibility that mycoplasmas possess the activity of TNF-alpha induction which might be responsible for a part of enhancement of cytotoxic activity of macrophages and resistance to infection with mycoplasmas in vivo.

Animals↗

Mycoplasmas induce transcription and production of tumor necrosis factor in a monocytic cell line, THP-1, by a protein kinase C-independent pathway.

We demonstrated that mycoplasmas (MP), previously shown to augment the antitumor activity of murine peritoneal macrophages, also induce cytotoxic activity in a human monocytic cell line, THP-1. THP-1 cells were induced to produce cytotoxic activity by MP in a time- and dose-dependent manner. By using neutralization by antibody against tumor necrosis factor (TNF), the cytotoxic activity was shown to be due to TNF released from the MP-stimulated cells. Studies with inhibitors of second-messenger pathways and Northern RNA blot analysis indicated that a Ca2(+)-dependent, but not protein kinase C-dependent, biochemical pathway is involved in MP-induced TNF production by THP-1 cells and that MP induce TNF production in the cells at the level of transcription. MP, unlike other bacteria, lack cell walls and lipopolysaccharide. The possible involvement of a TNF production mechanism distinct from that triggered by lipopolysaccharide is discussed.

Animals↗

The diagnosis of mild emphysema. Correlation of computed tomography and pathology scores.

Early and accurate diagnosis of emphysema is said to be invaluable for identification of clinically silent and mild emphysema. Recently, computed tomography (CT) has been much advocated for its efficacy in detailed diagnosis of emphysema, and the results have been compared with the pathology grade of emphysema in resected lung specimens. To assess the ability of high resolution CT scan in detecting and grading mild emphysema, we correlated the high resolution CT scan with the pathology grade of emphysema and the destructive index (DI) of lung specimens from 42 patients undergoing thoracotomy for a solitary pulmonary nodule. The high resolution CT scan and the cut surface of the lung, corresponding exactly to the same plane of the CT scan image, were assessed using the picture-grading system of Thurlbeck and coworkers on a scale of zero to 100. The CT scores for all patients ranged from 12 to 57, with a mean +/- SD of 22.1 +/- 9.6 using 1-mm collimation (n = 35), and from 7 to 46 with a mean +/- SD of 16.5 +/- 8.3 using 5-mm collimation (n = 33). The pathology scores ranged from 10 to 57, with a mean +/- SD of 23.2 +/- 9.8 (n = 42). The DI ranged from 15.4 to 67.1, with a mean +/- SD of 31.4 +/- 10.8 (n = 42). The CT scores using 1-mm and 5-mm collimation correlated significantly with the pathology scores (r = 0.68 and 0.76, respectively, p less than 0.001), and with the DI (r = 0.62 and 0.74, respectively, p less than 0.001). The pathology scores correlated significantly with the DI (r = 0.72, p less than 0.001). We therefore concluded that high resolution CT can help to identify the presence and grading of mild emphysema.

Adult↗

[Induction by Staphylococcus aureus L-form of tumor necrosis factor-alpha from macrophages].

Induction of tumor necrosis factor-alpha (TNF-alpha) by Staphylococcus aureus L-form was investigated. The supernatant of a macrophage culture mixed with S. aureus L-form showed a potent cytotoxic activity to L cells. Addition of anti TNF-alpha antibody inhibited completely the cytotoxic activity of the supernatant, indicating that the activity might be due mostly to TNF-alpha. To investigate localization of TNF-alpha production, the membranes of hypotonicity treated L-form were layered on a step-gradient composed of an upper and lower layers of 35% and 50% sucrose, respectively. The membranes were banded at the interface of 35% and 50% of sucrose. The activity of TNF-alpha production of the membrane fraction was 10-times higher than that of the soluble fraction.

Animals↗

[Membranous and respiratory bronchiole disease in patients with normal air-flow and mild air-flow limitation].

In order to evaluate the role of membranous (MB) and respiratory bronchioles (RB) in patients with normal and mild airflow limitation, we quantitatively assessed the pathological change of MB, RB and alveoli in the pathological specimens of 42 patients who required surgery for removal of a solitary pulmonary nodule. We also measured the destructive index (DI), to evaluate the destruction of alveolar walls. High resolution CT scan and pulmonary function tests were performed within 1 month of resection. Results of the pulmonary function tests correlated with pathological changes of RB better than with MB. Significant correlation between the emphysema score of resected lung and the pathological change of MB and RB was observed. There was also significant correlations between the CT score or DI and RB changes. However, there was no significant correlation between the CT score or DI and the change of MB. There were significant differences between smokers and nonsmokers in the results of pulmonary function test, pathological changes of RB and severity of emphysema. We postulate that RB is responsible for subtle functional abnormalities in the early stages of chronic obstructive pulmonary disease.

Adult↗

Active immunization against virus infections due to antigenic drift by induction of crossreactive cytotoxic T lymphocytes.

We have examined whether active immunization with c13 protein, a hybrid protein of the first 81 amino acids of the viral NS1 nonstructural protein and the HA2 subunit of A/PR/8 (H1N1) hemagglutinin, could protect BALB/c mice from challenge with A/PR/8 H1 subtype virus. Mice immunized with the c13 protein had a significant reduction of pulmonary virus titers with A/PR/8 (H1) virus, but failed to limit the replication of A/PC (H3) virus, which reflects the in vitro CTL activity of c13 immune spleen cells. We observed that the epitope recognized by HA2 specific CTL, which are induced by a derivative of c13 protein, is highly conserved among H1 and H2 subtype virus strains. This led us to test whether active immunization with c13 protein would also limit pulmonary virus replication in mice infected with the A/TW virus, a virus of the H1 subtype, which was isolated in 1986, and with a virus of the H2 subtype, A/Japan/305/57. Immunized mice had significantly lower lung virus titers than did control mice, and did not possess any neutralizing antibodies to the challenger viruses. These results indicate that active immunization with a fusion protein containing the cross-reactive CTL epitope protects mice from influenza infection by inducing CTL against influenza A H1 and H2 subtype virus strains, which markedly vary in their antibody binding sites on the HA1. The ability to induce active cross-reactive immunization with a fusion protein which contains a highly conserved CTL epitope offers a model for vaccine approaches against viruses which undergo significant variations in their antibody binding sites.

Animals↗

Cytotoxic T lymphocytes recognize a cross-reactive epitope on the transmembrane region of influenza H1 and H2 hemagglutinins.

A cross-reactive cytotoxic T lymphocyte clone was produced by stimulation with a hybrid protein that contained a portion of the NS1 and the HA2 subunit of A/PR/8/34 (H1N1) virus. Transfer of this clone clears virus from the lungs of mice challenged with H1 or H2 viruses. In these experiments we define the location of the protective CTL epitope to the transmembrane portion of the influenza A virus hemagglutinin which is well-conserved on H1 and H2 subtype viruses. The H1 and H2 cross-reactive CTL clone recognized a synthetic peptide of 23 amino acids (anchor peptide) corresponding to the transmembrane domain of the A/PR/8 (H1) HA as well as the comparable anchor peptide of the A/JAP (H2) HA. The anchor peptide of the A/PR/8 HA competed against the anchor peptide of A/JAP HA in cold target inhibition tests. These results indicate that the epitope recognized by the cross-reactive CTL is located on the transmembrane region of both A/PR/8 HA and A/JAP HA. We prepared synthetic peptides to define the epitope within the transmembrane region of A/PR/8 HA which is recognized by a cross reactive CTL clone. The results indicate that residues 518-528 in the transmembrane region of A/PR/8 HA contain the cross-reactive CTL epitope.

Amino Acid Sequence↗

[Induction by mycoplasmas of tumor necrosis factor-alpha from macrophages].

Several species of mycoplasmas including M. pneumoniae, the causative agent of human respiratory infection, were investigated for tumor necrosis factor-alpha (TNF-alpha) induction. The cytotoxic activity to Meth A cells of peritoneal macrophages purified from BALB/c mice was enhanced markedly when cultured with either viable or nonviable mycoplasmas. The supernatant of macrophage culture mixed with mycoplasmas, M. pneumoniae or A. laidlawii, showed a potent cytotoxic activity to TNF-alpha-sensitive but not to TNA-alpha-insensitive L cells. Addition of anti-TNA-alpha antiserum inhibited completely the cytotoxic activity of the supernatant, indicating that the cytotoxic activity is due mostly to TNF-alpha. These results strongly suggest that mycoplasmas possess an activity to induce TNF-alpha, which enhances the cytotoxic activity of macrophages and prevent infection with mycoplasmas in vivo.

Animals↗

[Comparison of high resolution computed tomography and pulmonary function tests in diagnosis of mild emphysema].

To assess the ability of high resolution CT scan and pulmonary function tests in detecting and grading mild emphysema, we correlated the high resolution CT scan and pulmonary function tests with the pathologic grade of emphysema and the destructive index of lung specimens from 42 patients undergoing thoracotomy for solitary pulmonary nodules. Using the high resolution CT scan, we could identify the pathologic grade of mild and moderate emphysema. By measuring diffusing capacity per unit alveolar gas volume (DLco/VA), it seemed to be possible to detect the mildest degree of alveolar destruction assessed by the destructive index, which was not detected by high resolution CT scan. The reason for these results seemed to be that we assessed the severity of emphysema by detecting the air space enlargement on high resolution CT scan images caused by the destruction of alveolar walls, which were detectable by measuring DLco/VA. We conclude that it is possible to detect mild emphysema using the combination of high resolution CT scan and pulmonary function tests.

Adult↗

[Membranous bronchiole diseases in patients without chronic air-flow limitation].

In order to investigate the relationship between pulmonary function and diseases involving membranous bronchioles (MB), we measured airway diameters and quantitated airway pathology scores in MB in 35 patients who required surgery for removal of a solitary nodule. The patients had FEV1 greater than 80% of the predicted value. Measurements of FEV1, nitrogen washout curve (delta N2/L), closing volume as a percentage of vital capacity (CV/VC%), and maximal flow rate at 50% of VC (V50) were made prior to the surgery. There were 21 smokers and 14 nonsmokers. There was 1.02 MB per cm2 of tissue. The mean internal diameter for MB cases was 0.55 +/- 0.05 mm. The total pathology scores for all MB cases were less than those found in North American populations. Inflammation scores were higher in patients with FEV1 less than 100% that of the predicted (p less than 0.05). FEV1 values correlated with goblet cell metaplasia (p less than 0.05). Tests specific for small airways did not correlate with the pathology scores in MB. We concluded. In 35 Japanese cases some diseases in MB were detected among cases whose average age was 59.5 years (42-78 yrs) and whose FEV1 was greater than 80% of the predicted value. The disease showed little effects of smoking and it was relatively minor in comparison with North American cases. This degree of abnormality is considered to be found in normal Japanese as well. It is supposed that an ordinary pulmonary function test would not reveal this abnormality in MB.

Adult↗

HA2 subunit of influenza A H1 and H2 subtype viruses induces a protective cross-reactive cytotoxic T lymphocyte response.

Influenza H1 subtype-specific CTL can be induced by secondary stimulation of a hybrid protein of the first 81 amino acids of the viral NS1 non-structural protein and the HA2 subunit of A/Puerto Rico/8/34(H1N1) hemagglutinin. In addition, a derivative of this protein with 65 amino acids deleted from the N-terminal end of HA2 can also generate H1 subtype-specific CTL in bulk cultures. CTL clones established by stimulation with the derivative protein demonstrated cross-reactive lysis of target cells infected with virus strains of the H1 and H2 subtypes. Cold target competition experiments with CTL clones as effectors demonstrated that the Ag specificity between these two hybrid proteins is identical. Adoptive transfer of the CTL clone significantly reduced virus titers in the lungs of mice infected with the virus strains of the H1 or H2 subtype but not those infected with the H3 subtype virus in vivo, which reflects the in vitro CTL clone activity. These experiments demonstrate that an epitope on the hemagglutinin that is conserved on virus strains of the H1 and H2 subtypes induces a protective CTL response. These results suggest an alternative approach for developing influenza vaccines by using conserved antigenic sites on the hemagglutinin HA2 subunit to avoid the problem of frequent antigenic mutations of the HA1 subunit antibody binding sites.

Animals↗

Induction of interferon production by natural killer cells by organogermanium compound, Ge132.

Interferon (IFN)-inducing activity of the organogermanium compound Ge132 in human peripheral mononuclear cells was investigated. By using Percoll discontinuous density gradient centrifugation, peripheral blood nonphagocytic and nonadherent mononuclear cells were divided into the low-and high-density fractions. Natural killer (NK)-enriched low-density fractions, but not the T-cell-enriched high-density fractions, showed IFN production by the stimulation of Ge132. The maximal titer of IFN by NK-enriched fractions (F1 + F2) was observed after a 74-h cultivation in the presence of 200 micrograms/ml Ge132. IFN production by the NK-enriched fractions was abrogated by treatment of the cells with monoclonal antibody against human NK cells in the presence of complement. The treatment with antiserum-neutralizing human IFN-gamma resulted in a marked reduction, indicating that a major part of IFN was IFN-gamma. These results suggested that Ge132 might possess affinity to NK cells, inducing IFN production by NK cells.

Antigens, Differentiation, T-Lymphocyte↗

Small airways disease in patients without chronic air-flow limitation.

We measured airway diameter and quantitated airway pathology scores in respiratory bronchioles (RB) and membranous bronchioles (MB) in 37 patients undergoing resection for solitary pulmonary nodules. The patients had FEV1 greater than 75% of the predicted. Measurements of FEV1, nitrogen washout curve (delta N2/L), closing volume as a percentage of vital capacity (CV/VC%), and maximal flow at 50% of VC (V50) were made prior to surgery. There were 22 smokers and 15 nonsmokers. There were more MB per cm2 of tissue than RB (0.74 versus 1.02). The mean internal diameter for MB was 0.55 +/- 0.27 mm; for RB it was 0.47 +/- 0.15 mm. There were more MB greater than 1.0 mm in internal diameter than RB greater than 1.0 mm, and the mean diameter of MB was less than that noted in autopsy specimens. The total pathology scores for all airways were less than those found in Caucasian populations. Pathology scores were higher for MB than for RB, except for pigment deposition. Inflammation scores were higher in patients with FEV1 less than 100% predicted both for RB (p less than 0.05) and for MB (p less than 0.05). FEV1 values correlated with goblet cell metaplasia scores (p less than 0.05) for MB and with fibrosis (p less than 0.05), pigment deposition (p less than 0.05), and intraluminal macrophages (p less than 0.05) for RB. Tests specific for small airways (delta N2/L, V50, CV/VC%) did not correlate with the pathology scores.

Adult↗