Biomedical subjects
T Yoshimatsu
Publications and source records attributed to T Yoshimatsu.
Esophagopleural fistula originating from diverticulum after pneumonectomy. A case report and review of the literature.
Esophagopleural fistula (EPF) is a very rare and fatal disease. A 56 year-old man developed a pyothorax resulting from an EPF with esophageal diverticulum after a right pneumonectomy. The patient was successfully treated with a three-stage operation and control of infection and nutritional status. First, fenestration was performed, then an ante-thoracic gastroplasty, and a radical thoracoplasty was performed. Surgical management, nutritional support, and control of infection were the cornerstones of the treatment of EPF.
Induction of Tumor-Specific Cytotoxic T Lymphocytes from Regional Lymph Node Lymphocytes of Human Breast Cancer.
BACKGROUND: In this study we activated breast cancer-specific cytotoxic T lymphocytes (CTL) from regional lymph node lymphocytes (RLNL) of HLA-A2-positive patients with breast cancer. METHODS: Freshly isolated RLNL were stimulated with solid phase anti-CD3 monoclonal antibody followed by expansion with recombinant interleukin-2. Subsequently, the RLNL were stimulated with an irradiated HLA 0201 breast cancer cell line,MCF-7, at a responder/stimulator ratio of 10/1 once a week for 2 weeks. RESULTS: The cultured RLNL exhibited specific lysis against MCF-7 in all 5 HLA-A2-positive patients tested, but not in 2 HLA-A2-negative patients. Cytotoxicityagainst MCF-7 was substantially inhibited by addition of anti-HLA-A2 mAb. In 3 of 5 HLA-A2-positive patients, anti-MCF-7 CTL also exhibited a substantial levelof reactivity against PC-9, an HLA-A0206-positive lung adenocarcinoma cell line. Conversely, anti -PC-9-specific CTL were inducible by multiple stimulations ofRLNL with PC-9 cells in 2 of 3 patients. CONCLUSION: These results suggest that several common tumor antigens might exist among HLA-A2-positive breast cancers, some of which may be shared with lung adenocarcinomas.
Improvement of retroviral packaging cell lines by introducing the polyomavirus early region.
To obtain high-titer recombinant retroviruses, we constructed plasmid pDL+, which carries the extended Psi region and the polyomavirus early region, including the replication origin and the early gene. Although pDL+ is useful for obtaining high-titer recombinant retroviruses, this vector plasmid is difficult to modify further for tissue-specific expression of foreign genes. To overcome this problem, the coding region of the polyomavirus early gene was expressed in the packaging cell lines. We modified the packaging cell lines, psi2 and PA317, by stably introducing the polyomavirus early gene, and established psiMP34 and psiMP37 from psi2, and PAMP51 from PA317. In the transient expression system using plasmids with and without the polyomavirus replication origin, the titers of recombinant retrovirus produced by these cell lines were 10-100 times higher than produced by the parent cell line and reached levels of 0.5-1.5 x 106 cfu/ml. Expression of the polyomavirus early gene in the packaging cell lines did not stimulate the production of replication-competent retrovirus. We also routinely established stable clones producing retrovirus titers over 1 x 10(7) cfu/ml from psiMP34 and PAMP51. We found that the activity of the long terminal repeat (LTR) promoter is stimulated by the polyomavirus early region protein(s) in these cell lines. Therefore, increases titer can be expected to occur in all the retroviral vectors in which LTR promoter is used to transcribe the retroviral genome.
Expression of the melanoma antigen-encoding gene in human lung cancer.
BACKGROUND AND OBJECTIVES: We surveyed expression of melanoma antigen-encoding genes in lung cancer because of promising implications for immunotherapy. METHODS: We studied 57 human lung carcinoma specimens using the reverse transcription-polymerase chain reaction (RT-PCR). RESULTS: In the samples, the expression of melanoma antigen-encoding genes 1, 2, and 3 was observed in 9/43 (20.9%), 13/43 (30.2%), and 22/48 (45.8%), respectively. In 28 cases in which all three messenger RNAs were sought, 18 (64.3%) showed expression of at least one gene, 10 (35.7%) showed expression of two or three genes, and 10 (35.7%) were negative for all three genes. In a clinicopathologic analysis, melanoma antigen-encoding genes 1 and 3 were frequently expressed in squamous cell carcinomas (P = 0.0543) and in cases with regional lymph node metastasis (P = 0.0572), respectively. CONCLUSIONS: The high incidence of melanoma antigen-encoding gene expression in lung cancer indicates the possibility of a future specific immunotherapy for this disease.
Transduction of glioma cells using a high-titer retroviral vector system and their subsequent migration in brain tumors.
The intracranial migration of transduced glioma cells was investigated in order to improve the treatment of malignant glioma by gene therapy using retroviral vectors. In this study, about half the volume of the tumor mass could be transduced in 14 days after only a single implantation of 3 x 10(5) retrovirus-producing cells into a tumor mass with a diameter of 5 mm. Moreover, we were able to follow the migration of glioma cells transduced by the lacZ-harboring retroviruses originating from the high-titer retrovirus-producing cells. Besides the importance of using a high-titer retroviral vector system, our results also indicate that the implantation site of the virus-producing cells and the interval between the implantation of the virus-producing cells and the subsequent administration of ganciclovir are important factors for the efficient killing of glioma cells.
Autologous tumor-specific cytotoxic T lymphocytes in a patient with lung adenocarcinoma: implications of the shared antigens expressed in HLA-A24 lung cancer cells.
Human lung adenocarcinoma-specific cytotoxic T lymphocytes (CTL) were generated by multiple stimulations with autologous tumor cells (named A110L) from regional lymph node lymphocytes and tumor-infiltrating lymphocytes expanded by solid-phase anti-CD3 monoclonal antibody (mAb) and recombinant interleukin-2. The CTL lysed A110L but failed to kill either autologous B lymphocytes immortalized by the Epstein-Barr virus or K562. The killing activity of the CTL against autologous A110L was inhibited by anti-MHC class I mAb (W6/32), but not by anti-MHC class II mAb. The CTL produced interferon-gamma and GM-CSF in response to A110L and the production was completely blocked by the addition of anti-MHC class I mAb. The HLA type of the CTL was HLA-A2/A24, B52/B54, Cw1/-. Allele-specific deletion of HLA-A2 molecules was observed in A110L by staining with anti-HLA-A2 mAb. A partial blocking effect on the cytokine production from the CTL was also obtained with anti-CD8, and anti-HLA-A24 mAbs, but not with anti-MHC class II, anti-CD4 and anti-HLA-A2 mAbs. To analyze further the mechanism of antigen recognition by the CTL, the cross reactivity of the CTL against several HLA-A locus-matched (HLA-A24+) and mismatched allogeneic tumor cells (HLA-A24-) was investigated. The A110L-specific CTL showed a weak but significant cytotoxicity against some HLA-A24 positive lung cancer cell lines, such as Sq-1 (HLA-A11/A24, squamous cell carcinoma) and PC-9 (HLA-A2/A24, adenocarcinoma), but failed to kill HLA-A locus-mismatched allogeneic tumors. This cross reactivity of the CTL against Sq-1 and PC-9 was blocked by anti-MHC class I mAb. These results thus demonstrate that shared common tumor antigens might exist among lung cancer cells in the context of HLA-A24.
Effects of interleukin-12 on the induction of cytotoxic T lymphocytes from the regional lymph node lymphocytes of patients with lung adenocarcinoma.
Lung cancer-specific cytotoxic T lymphocytes (CTL) were induced by repeated stimulations of regional lymph node lymphocytes (RLNL) in lung cancer patients with either autologous or HLA-A-locus-matched tumor cells. To investigate the effect of interleukin-12 (IL-12), IL-12 was added during the stimulation of RLNL from HLA A24/adenocarcinoma patients with either autologous tumor cells or HLA A24-positive adenocarcinoma cells (PC-9) in combination with, or instead of interleukin-2 (IL-2), and then the cytotoxic activity, cytokine production and populations of the lymphocyte subsets were examined. The addition of IL-12, or the substitution of IL-2 by IL-12 was found to enhance the cytotoxic activity and the cytokine production (IFN-gamma, GM-CSF) of the CTL as compared with IL-2 alone. The cytotoxic activity and cytokine production were both partially inhibited by anti-MHC-class I monoclonal antibody. The CTL thus induced by IL-12 had a higher proportion of CD3+/CD56+ cells than the CTL induced with IL-2 alone. The positively selected CD8+/CD56- lymphocytes showed PC-9-specific cytotoxic activity, because the population did not show any cytotoxicity to K562 or A549 (HLA-A26/A30). However, the CD3+/CD56+ lymphocytes were cytotoxic to both PC-9 and K562. In conclusion, IL-12 is considered to be a useful cytokine for both the induction of lung-cancer specific CTL and the augmentation of non-MHC-restricted cytotoxicity against tumor cells, and may be applicable for adoptive immunotherapy using CTL.
[A clinical study of causes of death from active pulmonary tuberculosis with chemotherapy].
We evaluated retrospectively the causes of death from active pulmonary tuberculosis by the review of records and chest radiograohs of 364 patients (male 282, female 82) with active pulmonary tuberculosis, who were admitted to our hospital during 1995 to 1998. 43 patients (male 33, female 10) were died under anti-tuberculous chemotherapy. 20 cases were tuberculous death; death from acute progression of tuberculosis without response to chemotherapy (acute progression group) in eight cases and death from debility in spite of partial response to chemotherapy (debility group) in eight cases. 23 cases were died from underlying diseases; death from malignant neoplasmas (malignant group) in nine cases and death from complication of bacterial pneumonia (pneumonia group) in seven cases. In acute progression group, the age (mean +/- SE) was 64.8 +/- 5.2 years old and the survival period from admission was 11.8 +/- 4.2 days. Five cases were laborer or unemployed. This group was characterized with far advanced diseases presenting extensive lung lesions complicated with DIC or hepatic dysfunction, low performance status (PS), severe malnutrition and lymphocytepenia. In debility group, the age was 70.8 +/- 3.9 years old and the survival period from admission was 254.6 +/- 90.7 days. Five cases were laborer or unemployed. This group was characterized with multiple underlying diseases, low PS, previous anti-tuberculous chemotherapy and resistance to INH and/or RFP. In malignant group, the age was 69.3 +/- 3.2 years old and the survival period from admission was 99.9 +/- 21.2 days. This group was characterized with relatively well nourished, relatively good PS in comparison with other groups, and lymphocytepenia. In pneumonia group, the age was 82.8 +/- 1.7 years old and the survival period from admission was 153.3 +/- 54.5 days. This group was characterized with remarkably advanced age, low PS related to underlying disorders of central nervous system. In the causes of death with active pulmonary tuberculosis under chemotherapy, inhomogenous groups were included. Extensive disease, low PS, malnutrition, lymphocytopenia, previous chemotherapy, resistance to INH and/or RFP, and poorer social circumstances seemed to be risk factors for tuberculous death. In contrast, underlying malignant nepolasma, lower PS, and far advanced age were seemed to be the risk factors for non-tuberculous death.
Prognostic value of the immunohistochemical detection of p16INK4 expression in nonsmall cell lung carcinoma.
BACKGROUND: The product of the p16INK4/CDKN2/MTS1 (p16) controls the transition from the G1 phase to the S-phase in the cell cycle by inhibiting the phosphorylation of the retinoblastoma gene product. A lack of p16 expression has been reported in various cancer cell lines and tumors; however, there have been only a few reports on the prognostic significance of p16 alteration. The authors studied p16 expression in nonsmall cell lung carcinoma (NSCLC) and also examined its correlation with clinicopathologic features and prognosis. METHODS: p16 expression was determined by immunohistochemical analysis of 115 paraffin specimens of primary NSCLC that were curatively resected. The immunohistochemical study was performed using the labeled streptavidin-biotin method with anti-p16 rabbit polyclonal antibody. RESULTS: Thirty-one of 115 NSCLC specimens (27%) showed negative p16 staining. The frequency of negative p16 expression was significantly higher in squamous cell carcinoma (39.5%) than in adenocarcinoma (20.3%) (P = 0.026). There were no statistically significant differences in the p16 status with respect to age, gender, smoking history, histologic differentiation, or stage of the disease. The Kaplan-Meier survival curves demonstrated that patients with negative p16 expression survived for a significantly shorter period of time than those with positive p16 expression (P = 0.043). p16 status was a significant prognostic factor, especially in patients with early stage disease (Stages I-II) (P = 0.039). CONCLUSIONS: A lack of p16INK4 expression in NSCLC was observed more frequently in squamous cell carcinoma than in adenocarcinoma, and also was found to be closely related to prognosis, especially in patients with early stage squamous cell carcinoma.
Thermostable alkaline cellulase from an alkaliphilic isolate, Bacillus sp. KSM-S237.
Thermostable alkaline cellulase (endo-1,4-beta-glucanase, EC 3.2.1.4) activity was detected in the culture medium of a strictly alkaliphilic strain of Bacillus, designated KSM-S237. This novel enzyme was purified to homogeneity by a two-step column-chromatographic procedure with high yield. The N-terminal amino acid sequence of the purified enzyme was Glu-Gly-Asn-Thr-Arg-Glu-Asp-Asn-Phe-Lys-His-Leu-Leu-Gly-Asn-Asp-Asn-Val- Lys-Arg. The enzyme had a molecular mass of approximately 86 kDa and an isoelectric point of pH 3.8. The enzyme had a pH optimum of 8.6-9.0 and displayed maximum activity at 45 degrees C. The alkaline enzyme was stable up to 50 degrees C and more than 30% of the original activity was detectable after heating at 100 degrees C and at pH 9.0 for 10 min. The enzyme hydrolyzed carboxymethylcellulose, lichenan (beta-1,3;1,4-linkage), and p-nitrophenyl derivatives of cellotriose and cellotetraose. Crystalline forms of cellulose (Avicel and filter paper), H3PO4-swollen cellulose, NaOH-swollen cellulose, curdlan (beta-1,3-linkage), laminarin (beta-1,3;1,6-linkage), and xylan were barely hydrolyzed at all.
Postoperative prognosis in patients with non-small cell lung cancer with synchronous ipsilateral intrapulmonary metastasis.
BACKGROUND: Non-small cell lung cancer with intrapulmonary metastasis (PM) was recently reclassified according to the revision of the TNM classification. To determine whether the new staging system is appropriate, we analyzed the postoperative prognosis of patients with synchronously detected and resected PM lesions. METHODS: Of 509 patients with non-small cell lung cancer who underwent surgical resection, 42 patients were revealed to have synchronous and ipsilateral PM. Their survival was compared with that of matched stage groups (without PM) by Kaplan-Meier test and log rank test. RESULTS: Two patients who were classified as stage I survived 40 and 30 months after operation, respectively. One patient was determined to be stage II, and survived 100 months postoperatively. Thirty-eight patients were classified as stage IIIA/IIIB (19 each) (90.5% of all cases with PM). There was no significant difference between 3- and 5-year survival rates of the PM stage IIIA group (34.2% and 34.2%) and those of the other IIIA (144 patients; 37.9% and 31.6%). Survival rates of such stage IIIA subgroups as PM, T3 and N2, were comparable. No significant differences were observed between the 3- and 5-year survival rates of the PM stage IIIB (16.6/16.6%) and those of the other stage IIIB (45 cases; 11.7% and 0.0%). The survival rates of such stage IIIB subgroups as PM, T4 and N3 were also similar. CONCLUSIONS: The new staging system for patients with synchronous resectable PM appears to be reasonable regarding survival. Most cases of PM are categorized as locally advanced disease; however, stage IIIA/IIIB cases have become a more heterogeneous population.
Treatment strategy for patients with surgically discovered N2 stage IIIA non-small cell lung cancer.
BACKGROUND: The treatment strategy for patients with non-small cell lung cancer and clinically negative, but surgically detected mediastinal lymph node metastasis (surgically discovered N2 disease) is controversial. METHODS: From August 1979 through December 1994, 53 patients with non-small cell lung cancer were found to have surgically discovered N2 disease. We retrospectively studied the clinical characteristics and the factors that influenced the prognosis in these patients. RESULTS: The 3-year and 5-year survival rates and the median survival for the 53 patients with surgically discovered N2 disease were 44%, 21%, and 26 months. Two thirds of the patients had adenocarcinoma. Only complete resection affected long-term survival; adjuvant therapy had no effect on survival. In regard to lymph node status, a single metastatic focus in the aortic area was associated with long-term survival. CONCLUSIONS: Patients with adenocarcinoma may require histologic determination of N2 disease. Complete resection, including extensive and complete mediastinal lymph node dissection, is warranted in patients with surgically discovered N2 disease. In particular, when the aortic lymph node (including stations 5 and 6) alone is involved, the patients should undergo as complete a resection as possible.
Optimal dose of basic fibroblast growth factor for long-segment orthotopic tracheal autografts.
When a primary anastomosis of the trachea is not feasible, extensive grafting is required. However, despite the use of omental wrapping for revascularization, long-segment tracheal grafts frequently do not maintain structural integrity because of insufficient blood supply. We examined the use of basic fibroblast growth factor for preservation of long-segment tracheal autografts after orthotopic transplantation with omental wrapping in 23 dogs. All animals received orthotopic tracheal transplantation, with 14-ring autografts that occupied a major part of the thoracic trachea, and omental wrapping. The 23 animals were classified randomly into six groups as follows: no treatment (group I, n = 3), topical administration of fibrin glue alone (group II, n = 4), fibrin glue enriched with 1 microg/cm2 basic fibroblast growth factor (group III, n = 4), fibrin glue enriched with 5 microg/cm2 basic fibroblast growth factor (group IV, n = 4), and fibrin glue enriched with 10 microg/cm2 basic fibroblast growth factor (groups V and VI, each n = 4). The omentum that was used to wrap the autografts was fed by the right gastroepiploic artery in groups I to V and by both the right gastroepiploic artery and splenic artery in group VI. All autografts in groups I and II showed dissolution. Ten of 12 autografts in groups III, V, and VI did not maintain long-term structural integrity. By contrast, all autografts in group IV showed long-term viability, as demonstrated by graft patency, epithelialization, cartilage morphology, and vascularity. We conclude that treatment with fibrin glue enriched with 5 microg/cm2 basic fibroblast growth factor in combination with omental wrapping may prolong the viability of long-segment tracheal autografts.
Combined carboplatin and cisplatin therapy in patients with advanced non-small cell lung cancer.
Combining cisplatin and carboplatin may eliminate some of the toxic effects of each agent and permit the use of higher doses, because these agents have different pharmacodynamics and dose-limiting toxicities. We investigated the safety and efficacy of these agents in combination. The toxicity profile was evaluated in a Phase I trial in 18 patients with advanced non-small cell lung cancer (NSCLC). Carboplatin was administered in doses ranging from 200 to 400 mg/m2 on day 1 and cisplatin at a dose of 80 mg/m2 on day 3. Only one cycle of chemotherapy was administered. Thrombocytopenia was the dose-limiting toxic effect. The maximal tolerated dose of carboplatin was 350 mg/m2. We then investigated the efficacy of the optimal dose of this combined chemotherapy in a Phase II trial in 13 patients. We used a carboplatin dose of 300 mg/m2 for safety in the Phase II trial. Three of 13 patients developed grade 3-4 hematologic toxicity, which was improved without major complications. A partial response was observed in 5 of 13 patients (38.5%). Combination chemotherapy with carboplatin (day 1, 300 mg/m2) and cisplatin (day 3, 80 mg/m2) showed promising effects in patients with advanced NSCLC.
Retroviral introduction of the p16 gene into murine cell lines to elicit marked antiproliferative effects.
The p16 gene is a candidate tumor suppressor, because mutation of the gene has been reported in many transformed cell lines and some primary tumor tissues. We have examined this possibility in murine cell lines (NIH3T3 and RSV-M) which lack p16 gene expression. Full-length human p16 cDNA was obtained from a HeLa cell line using polymerase chain reaction amplification. We constructed two separate retrovirus vectors carrying this p16 cDNA. First, we transduced the p16 cDNA into the murine cell lines using a retrovirus vector harboring the neomycin-resistance gene. The p16 gene-transduced cells formed no colonies after selection with G418, in contrast to the vector-transduced cells. Next, we used another retrovirus vector that expresses both the p16 cDNA and the Lac Z gene, which enabled us to distinguish affected cells from unaffected ones. Proliferation of the p16 gene-transduced cells was markedly inhibited and morphological change in the cells was also observed. Thus, we concluded that the p16 gene has an antiproliferative effect on the cell cycle and that the loss of its function may play a major role in dysregulated proliferation of the cells.
The induction of cytotoxic T lymphocytes against HLA-A locus-matched lung adenocarcinoma in patients with non-small cell lung cancer.
To induce cytotoxic T lymphocytes (CTL) against non-small cell lung cancer (NSCLC) efficiently, the induction of CTL was attempted using HLA-A locus-shared allogeneic NSCLC cells. T cells derived from either tumor tissue specimens or the regional lymph nodes of patients with NSCLC were stimulated twice or three times with an HLA-A2/A24-positive NSCLC cell line (PC-9), and thereafter the cytotoxic activity was examined by 51Cr-release assay. In patients with HLA-A24/ adenocarcinoma, anti-PC-9 cytotoxicity was induced in all 6 patients tested. Anti-PC-9 cytotoxicity was induced in 2 out of 5 patients with HLA-A2 (A24-)/adenocarcinoma, in 2 out of 4 patients with HLA-A24/squamous cell carcinoma, and 1 of 2 patients with HLA-A2/squamous cell carcinoma. The cytotoxic activity was observed to kill PC-9 selectively, not other NSCLC lines, and the activity was substantially blocked by anti-MHC class I antibody, but not by anti-MHC class II antibody. The PC-9-specific CTL produced gamma-interferon in response to autologous tumor cells. These results indicated that the anti-PC-9 cytotoxicity was mediated by cytotoxic T lymphocytes that may recognize the T cell epitope(s) shared and presented by HLA-A2 and/or HLA-A24-positive NSCLC.
[Treatment with expandable metallic stent in patients with carcinomatous airway stenosis--evaluation of patient's quality of life].
The expandable metallic stent was used in 10 patients with carcinomatous airway stenosis. In all patients, the respiratory symptoms improved immediately after insertion of the stent. Eight of the 10 patients, performance status improved. Totally, the use of expandable metallic stent improved the quality of life for patients with carcinomatous airway stenosis. Complication was minimal. We conclude that expandable metallic stent for treatment of carcinomatous airway stenosis are useful in emergent cases.