Quantitative analysis of the usage of human T-cell receptor alpha and beta chain variable regions by reverse dot-blot hybridization.
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Base Sequence↗
Biomedical subjects
T Matsutani
Publications and source records attributed to T Matsutani.
Polyclonal expansion of TCRBV2- and TCRBV6-bearing T cells in patients with Kawasaki disease.
We examined T-cell receptor (TCR) usage, cytokine production and antibody responses to superantigens in patients with Kawasaki disease (KD) to facilitate a better understanding of the immunopathogenesis of KD. The mean percentage of VB2- or VB6. 5-bearing T cells in peripheral blood mononuclear cells (PBMC) of patients with acute-phase KD was significantly higher than that of patients in the convalescent phase of KD or in healthy donors. Expansion of VB2- or VB6.5-bearing T cells was polyclonal because DNA sequences in the complementarity determining region 3 of VB2- and VB6.5-positive cDNA clones were all different from each other. The plasma levels of interleukin (IL)-1beta, IL-2, IL-6, IL-8, IL-10, interferon-gamma (IFN-gamma), tumour necrosis factor-alpha (TNF-alpha) and granulocyte colony-stimulating factor (G-CSF) were elevated in the acute phase of KD. We previously reported that streptococcal pyrogenic exotoxin C (SPEC) was a potent stimulator of VB2- and VB6.5-positive T cells and, furthermore, serum levels of anti-SPEC antibodies were significantly higher in patients with acute and convalescent KD than in age-matched controls. The results of the present study, together with those of our previous report, suggest that SPEC induces activation and polyclonal expansion of VB2- and VB6.5-positive T cells, and that SPEC-induced activation of T cells may lead to the pathogenesis of KD.
Acute Disease↗
Detection of T cell apoptosis after major operations.
OBJECTIVE: To detect T cell apoptosis in reduced peripheral lymphocyte counts in patients having major operations. DESIGN: Prospective study. SETTING: University hospital, Japan. SUBJECTS: 11 patients having oesophagectomy and 5 having laparoscopic cholecystectomy. INTERVENTIONS: To investigate T cell apoptosis we detected DNA fragmentation using electrophoresis, and T-cell receptor-gamma (TCR-gamma) gene amplification using polymerase chain reaction (PCR) in serum. MAIN OUTCOME MEASURES: Peripheral lymphocyte count and DNA extracted from the serum preoperatively and on postoperative days 1, 3, 5, and 7. RESULTS: The lymphocyte count decreased significantly until day 5 and then increased in the patients who had had oesophagectomy. DNA fragmentation and PCR products for the TCRgamma variable region gene were found in the serum DNA of 10 patients until day 5. No DNA fragmentation or PCR products were found in the serum of patients who had had laparoscopic cholecystectomy. CONCLUSION: These results suggest that transient T cell apoptosis occurs after major operations.
Adult↗
Glucocorticoid attenuates a decrease of antithrombin III following major surgery.
BACKGROUND: Major surgery, such as esophagectomy, activates inflammatory responses and the coagulation system, and this activation is characterized by release of inflammatory cytokines and a decrease in antithrombin-III (AT-III), respectively. Preoperative glucocorticoid administration has been reported to suppress circulatory cytokine levels after major surgery. PATIENTS AND METHODS: A total of 28 patients underwent esophagectomy for esophageal carcinoma; 14 of them were given 10 mg/kg of methylprednisolone intravenously upon induction of anesthesia and 14 served as controls. Circulating levels of tumor necrosis factor-alpha (TNF-alpha), interleukin 6 (IL-6), polymorphonuclear (PMN) elastase, thrombin-antithrombin III complex (TAT), AT-III, and albumin were measured before and immediately after the operation and on postoperative days (PODs) 1, 3, 5, and 7. RESULTS: TNF-alpha, IL-6, and TAT levels significantly increased after esophagectomy in both groups. AT-III and albumin decreased to their minimum levels on POD 1 and POD 3, respectively. Methylprednisolone treatment effectively inhibited the increases in TNF-alpha and IL-6 and the decreases in AT-III and albumin, but did not inhibit the increases in PMN-elastase and TAT levels. There were significant correlations between AT-III, IL-6, and albumin levels. CONCLUSIONS: These results suggest that methylprednisolone pretreatment attenuates the decrease in AT-III by reducing IL-6 production postoperatively.
Aged↗
Establishment and characterization of Japanese encephalitis virus-specific, human CD4(+) T-cell clones: flavivirus cross-reactivity, protein recognition, and cytotoxic activity.
We analyzed the CD4(+) T-lymphocyte responses of two donors who had received Japanese encephalitis virus (JEV) vaccine 6 or 12 months earlier. Bulk culture proliferation assays showed that peripheral blood mononuclear cells (PBMC) responded to JEV antigens (Ag) but also responded at lower levels to West Nile virus (WNV) and dengue virus type 1, 2, and 4 (D1V, D2V, and D4V, respectively) Ag. Five JEV-specific CD4(+) human T-cell clones and one subclone were established from PBMC of these two donors. Two clones responded to WNV Ag as well as to JEV Ag, whereas the others responded only to JEV Ag. Three of five CD4(+) T-cell clones had JEV-specific cytotoxic activity and recognized E protein. The HLA restriction of the JEV-specific T-cell clones was examined. Three clones were HLA-DR4 restricted, one was HLA-DQ3 restricted, and the HLA restriction of one clone was not determined. T-cell receptor analysis showed that these clones expressed different T-cell receptors, suggesting that they originated from different T lymphocytes. These results indicate that JEV vaccine induces JEV-specific and flavivirus-cross-reactive CD4(+) T lymphocytes and that these T lymphocytes recognize E protein. The functions and HLA restriction patterns of these T lymphocytes are, however, heterogeneous.
CD4-Positive T-Lymphocytes↗
[A case report of esophageal carcinoma with grade 3 response by preoperative chemotherapy using CDDP/5-FU/LV].
A 67-year-old man was diagnosed as having a type 3 advanced esophageal carcinoma by barium swallow and endoscopy. Biopsy specimens showed well-differentiated squamous cell carcinoma with positive immunostaining for p53, C-erb B-2 and negative for bcl-2. Two courses of chemotherapy using 5-FU, leucovorin and CDDP were performed before operation. Because no cancer cells were present in the surgical specimens, the effect was evaluated as grade 3. This neoadjuvant chemotherapy may be effective for esophageal carcinoma with a possible apoptosis mechanism.
Aged↗
Quantitative analysis of the usage of human T cell receptor alpha and beta chain variable regions by reverse dot blot hybridization.
We previously developed an adaptor ligation-mediated PCR method to amplify the T cell receptor (TCR) cDNA pools. In the present study we applied reverse dot blot hybridization to PCR-amplified specimens for quantitative analysis of the usage of TCR alpha and beta chain variable (V) region. 44 VA sequence-specific oligonucleotide probes (SSOPs) and 38 VB SSOPs were synthesized corresponding to unique sequences of VA and VB subfamilies. Peripheral blood lymphocytes of ten healthy donors and five T cell clones established from bone marrow cells were examined for VA and VB usage using this method. The results were consistent with those obtained by a colony hybridization method and those by immunofluorescence staining using monoclonal antibodies to VA and VB. Thus, reverse dot blot hybridization for TCR V(alpha) and Vbeta is a new, easy and dependable technique useful for analysis of VA and VB usage by human T cells.
Base Sequence↗
Accumulation and depuration of tributyltin oxide and its effect on the fertilization and embryonic development in the pacific oyster, Crassostrea gigas.
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Animals↗
Endoscopic dexamethasone injection following balloon dilatation of anastomotic stricture after esophagogastrostomy.
BACKGROUND: Anastomotic stricture is common after esophagogastrostomy. Recent advances in nonsurgical treatment include the silicon bougie and balloon dilatation. However, simple dilatation alone with a silicon bougie or endoscopic balloon dilator was repeated a mean of 4.7+/-5.4 times to control anastomotic stricture because of its temporary effect. METHODS: For 11 patients, endoscopic injection of dexamethasone (8 mg) around the anastomosis was done immediately after balloon dilatation (40 psi for 5 minutes). RESULTS: This method significantly reduced the number of the dilatations to 1.1+/-0.3 (P < 0.05). Ten of the 11 patients did not need any further treatment. There were no side effects or complications of dexamethasone injection. CONCLUSION: A combination of endoscopic balloon dilatation and dexamethasone injection provided an easy and safe method for preventing the recurrence of anastomotic stricture.
Anastomosis, Surgical↗
Non-restricted T cell receptor (TCR)-V alpha and -V beta gene usage in patients with pulmonary sarcoidosis.
Sarcoidosis is a systemic granulomatous disease of unknown etiology characterized by the pronounced accumulation of CD4+ T cells and macrophages in the affected organs. TCR variable (V) alpha and V beta gene usage in patients with sarcoidosis is still a matter of discussion. In this investigation, we analysed TCR-V alpha and -V beta gene usage in bronchoalveolar lavage fluid (BALF) and peripheral blood mononuclear cells (PBMC) of 30 patients with active pulmonary sarcoidosis using an adapter ligation method, reverse transcriptase-polymerase chain reaction (RT-PCR), and sequence-specific oligonucleotide probe (SSOP) analyses. There was no significant difference in TCR-V alpha or -V beta gene usage between BALF (n = 12) or PBMC (n = 27) of patients and PBMC of healthy subjects (n = 10). Neither selective TCR-V alpha nor -V beta expansion was observed in the paired BALF and PBMC from seven of nine patients. However, selective expansions were observed in a few TCR-V alpha or -V beta subsets in the BALF or PBMC of some individuals. Although a modest increase in a few TCR-V alpha or -V beta subsets was observed in the BALF or PBMC of some individuals, the increased TCR-V alpha or -V beta subsets were not closely associated with the HLA-DRB1, DQA1, DQB1, and DPB1 alleles of these patients. These results suggest that TCR-V alpha or -V beta gene usage is not restricted in both lung and peripheral blood in the majority of patients with active pulmonary sarcoidosis.
Adult↗
5-Hydroxytryptamine1B and alpha 2 adrenaline receptors in the brain of rats prenatally treated with methylazoxymethanol.
Methylazoxymethanol (MAM)-induced cerebral hypoplasia resulted in a significant increase in concentrations of 5-hydroxytryptamine (5-HT, serotonin) and norepinephrine in the frontal cortex, suggesting that these monoaminergic neurons were compressed due to smaller brain volumes. The serotonergic and noradrenergic presynaptic autoreceptors in rat brain are thought to be 5-HT1B receptors and alpha 2-adrenoceptors, respectively. If so, prenatal MAM treatment should increase the density of 5-HT1B and adrenaline alpha 2 receptors in the brain via the compression of noradrenergic and serotonergic axon terminals in the brains of rats with MAM-induced microencephaly. However, neither the densities nor the affinities of 5-HT1B and adrenaline alpha 2 receptors were changed in the MAM rats, suggesting that these presynaptic autoreceptors comprise only a small percentage of the total receptor population. Most 5-HT1B and adrenaline alpha 2 receptors were localized post-synaptically.
Alkylating Agents↗
Changes in urinary nitrate and nitrite during treatment of ulcerative colitis.
The urinary excretion rates of nitrate (NO3) and nitrite (NO2) were monitored in 14 patients with active ulcerative colitis during treatment using hydrocortisone and sulfasalazine. During the active phase of the disease, the NO3 excretion was significantly higher in the patients than in healthy controls (n = 6, p < 0.05), although it varied considerably among the patients. During the healing phase, the NO3 excretion decreased concurrently with improvement of symptoms and colorectal ulceration, but the NO2 excretion increased. During the inactive phase of the disease, the NO3 and NO2 excretions were significantly lower than during the active phase, and the NO2/NO3 ratio resembled that in the healthy controls. In contrast, a patient who failed to respond to treatment showed continuously high NO3 and NO2 excretion rates. These results indicate that urinary NO3 and NO2 excretions vary with the disease state in ulcerative colitis.
Adult↗
Increases in pulmonary artery pressure and cardiac output due to the inhibition of nitric oxide synthesis during operative stress.
Pulmonary artery pressure (PAP), cardiac output (CO), and urinary nitrate, a stable endproduct of nitric oxide (NO), were measured pre- and postoperatively in eight patients who underwent esophagectomy for squamous cell carcinoma of the thoracic esophagus. A significant elevation of PAP and CO on the day of operation (POD 0) was accompanied by a low concentration of urinary nitrate. A reduction in PAP and CO, and an increase in nitrate to the preoperative levels, were found on PODs 2 and 3, respectively, but urinary nitrate decreased again after POD 3. Consequently, the changes in PAP and CO were closely correlated with the nitrate concentration. These results suggest that operative stress inhibited NO synthesis with a transitory induction of endogenous NO synthesis postoperatively.
Aged↗
Molecular analysis of T cell receptor V beta chain to detect leukemia cell clonality in patients by adaptor ligation-mediated polymerase chain reaction.
We have developed a simple and rapid method to analyze the clonality of leukemia cells. After three rounds of amplification by adaptor-ligation polymerase chain reaction (PCR), the cDNA is cut with AluI, HaeIII, RsaI, and Sau3AI, and analyzed by polyacrylamide gel electrophoresis. The size of the restriction fragments is compared to that of the published restriction fragments size each TCR-beta subfamily V region. The sensitivity of adaptor-ligation PCR restriction enzyme analysis (AL-PCR-REA) was 10(-4) MOLT-4 T-ALL cell population in the normal peripheral blood lymphocytes (PBL). Application of AL-PCR-REA to PBL and bone marrow (BM) cells from eight clinical leukemia samples indicated that a detection sensitivity was rather low, but revealed the clonality of all eight clinical samples. This AL-PCR-REA method can detect clonality without the need for either radioisotopes or sequencing procedures.
Base Sequence↗
Transsternal thoracotomy for bilateral pulmonary metastasis.
Advances in the surgical treatment of primary malignancies and the recent chemotherapy have led to an expansion of the surgical treatment of metastatic lung tumors. However, multiple pulmonary metastases are often found and may affect both lungs. It is difficult to reach tumors in the posterior parts of the lung when using a common midsternal approach, especially lesions located in the left lower lobe. We performed transsternal simultaneous bilateral thoracotomy on 10 patients with bilateral lung tumors (9 bilateral metastatic pulmonary tumors and 1 bilateral primary lung cancer). This procedure provides a wide operative field and is an effective method of thoracotomy for patients with bilateral lung tumors. In future, this method should be more actively performed for patients in whom it is indicated.
Adolescent↗
Excitatory amino acid receptors in brains of rats with methylazoxymethanol-induced microencephaly.
We used methylazoxymethanol-acetate (MAM), a potent alkylating agent, to produce microencephaly in offspring by injecting it into pregnant rats on day 15 of gestation. Binding activities of central excitatory amino acid receptors were examined in Triton-treated membranes prepared from brains of adult offspring with MAM-induced microencephaly (MAM rats). MAM rats exhibited approximately 40-50% reductions of the wet weights of the cerebral cortex, hippocampus and striatum compared to those in controls. In the cortex and hippocampus of MAM-rats, total bindings of [3H]glutamate (Glu) (which is sensitive to N-methyl-D-aspartate (NMDA) receptor), and strychnine-insensitive [3H]glycine (Gly) and (+)-5-[3H]methyl-10,11-dihydro-5H-dibenzo[a,d]cyclohepten-5,10-imi ne (MK-801; a noncompetitive antagonist of NMDA receptor), were reduced to approximately 40% of those in controls. Similarly, in both regions of MAM rats, total bindings of [3H]kainate and DL-alpha-amino-3-[3H]hydroxy-5-methylisoxazole-4-propionic acid (an agonist of quisqualate receptors), were reduced to approximately 35-50% of those in controls. However, total bindings of these radioligands in the striatum of MAM rats were more than 65% of those in controls, despite the significant loss of striatum mass. However, specific bindings of radioligands in the striatum of MAM rats were elevated by more than 60% of those in controls, and Scatchard analysis revealed that elevations of [3H]Glu, [3H]Gly and [3H]MK-801 bindings were due to a significant increase in the densities of binding sites, with their affinities remaining unaltered. Spatial recognition ability examined by an 8-armed radial maze task was markedly impaired compared to those in controls. These results suggest that the proliferation of neurons bearing excitatory amino acid receptors (EAA) in the striatum is less affected by MAM treatment on day 15 of gestation than that in the cortex and hippocampus in spite of drastic weight loss in these brain regions. The significant reduction of EAA receptors in the cortex and hippocampus may be involved in the impairment of spatial memory observed in MAM-treated rats.
Animals↗
A case of trans-sternal bilateral thoracotomy for bilateral lung cancer.
Trans-sternal bilateral thoracotomy was performed to resect the right upper lobe and the left S1 + 2 + S3, and to complete lymphadenectomy in a 35-year-old female case of lung cancer in whom multiple lesions were suspected. Trans-sternal bilateral thoracotomy was considered to be useful for one-stage surgery in patients in whom bilateral lung cancer is suspected or confirmed, because it provides a sufficient surgical field enabling the resection of lung and lymph nodes. This may be the first case report of trans-sternal bilateral thoracotomy to treat multiple primary lung cancer.
Adenocarcinoma↗
[Study on cases of resected primary lung cancer in young persons].
A clinical study was conducted of 17 patients aged less than 40 years who received resection for lung cancer in our department. The 17 cases made up 1.8% of the total series of 924 resected lung cancer cases, with the number of cases increasing as the age of 40 years was approached. The male-female ratio was 1.1:1, with proportion of women higher than in the total series of lung cancer cases. The histological type of included a high proportion of adenocarcinomas (47.0%), while squamous cell carcinomas were few. In addition, the proportion of tumors of low-grade malignancy such as carcinoid tumors and mucoepidermoid carcinomas was high. The majority (58.8%) of cases were detected by mass screening. As a result, the number of stage I cases was high (10 cases, 58.8%), and curative resection could be performed in 70.8%. The prognosis of these young patients did not differ significantly from that of the total resected group, with a 5-year survival rate of 62.4% achieved. It was considered that the prognosis of lung cancer in young persons can also be improved with early detection by mass screening and active surgical intervention.
Adenocarcinoma↗