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Biomedical subjects

T Shimano

Publications and source records attributed to T Shimano.

At least 37 records · Page 2Linked to original sources
PubMed1997

Augumentation of splenic antitumor immunity by local immunotherapy in gastric cancer patients.

We previously reported that the antitumor effect of OK-432, a streptococcal preparation, was markedly augmented when this agent was injected into tumors together with fibrinogen. In order to elucidate the effect of this treatment on the spleen, we assessed splenic function in gastric cancer patients receiving preoperative local immunotherapy with OK-432 and fibrinogen. Immunohistochemical studies of the spleen at 7 days after intratumoral injection therapy revealed numerous macrophages phagocytizing OK-432 in the splenic sinuses. Phenotypic analysis of splenocytes by flow cytometry revealed an increase in the CD4/CD8 ratio and in the expression of HLA-DR, CD25, and Leu M3 by splenic T cells of the patients treated with OK-432 plus fibrinogen when compared to patients treated with OK-432 alone or untreated patients. Splenic T cells from patients treated with OK-432 plus fibrinogen showed significantly higher cytotoxicity against Daudi and K562 cells than T cells from control patients (p < 0.05), and culture of these splenic T cells with recombinant IL-2 induced the expansion of lymphokine-activated killer cells. These results demonstrate that local immunotherapy with a mixture of OK-432 and fibrinogen effectively augumented splenic antitumor immunity in gastric cancer patients.

Antineoplastic Agents↗
PubMed1997

A detailed analysis of the role of K-ras gene mutation in the progression of colorectal adenoma.

To elucidate the role of ras gene mutations during the early stage of colorectal tumour progression, K-ras gene mutations were analysed in 32 benign adenomas and 36 adenomas with focal carcinoma in the colorectum by microscraping of histologically pure regions from tissue sections, polymerase chain reaction-restriction fragment length polymorphism and in part by direct sequencing. Several regions were scraped out and analysed when an adenoma contained areas with different grades of dysplasia. The frequencies of K-ras gene mutation in mild dysplasia, moderate dysplasia and focal carcinoma were 19% (7/36), 51% (25/49) and 39% (14/36) respectively. The K-ras gene status was heterogeneous in 4 of the 11 benign adenomas from which multiple samples were obtained, and mutations were always found in the regions with more advanced dysplasia in these adenomas. Thirteen of the 36 adenomas with focal carcinoma showed heterogeneity of mutations between the adenoma region and the focal carcinoma. Seven of which had mutations only in the adenoma region. These findings indicated that the K-ras gene mutations occur during the late stage of adenoma progression and may confer a more advanced morphological phenotype of adenoma, but these mutations are not mainly involved in malignant transformation from adenoma to carcinoma.

Adenoma↗
PubMed1997

Successful endoscopic injection sclerotherapy with N-butyl-2-cyanoacrylate following the recurrence of bleeding soon after endoscopic ligation for ruptured duodenal varices.

Bleeding from duodenal varices in a 63-yr-old man with alcoholic cirrhosis of the liver was found at endoscopy, and ligation surgery was carried out. Ten months after the operation, bleeding from the duodenal varices occurred and was treated by endoscopic ligation. However, after performing this procedure, bleeding again occurred 1 week later. Hemostasis was finally achieved by endoscopic injection sclerotherapy with N-butyl-2-cyanoacrylate. For the 22 months since the injection, the patient has been free from further bleeding. These results suggest that endoscopic injection sclerotherapy with N-butyl-2-cyanoacrylate was effective in bringing about immediate cessation of the bleeding in duodenal varices and that long-term hemostasis can be expected.

Duodenoscopy↗
PubMed1996

Analysis of cytotoxic activity of the CD4+ T lymphocytes generated by local immunotherapy.

We previously reported that the anti-tumour effect of OK-432 is considerably enhanced by its intratumoral injection together with fibrinogen. In the present study, we generated killer T cells by culturing tumour-infiltrating lymphocytes from thyroid cancer patients who had received this local immunotherapy. Phenotypic analysis revealed that the T cells were positive for CD3+, CD4+, Leu8-, CD45RO+ and T-cell receptor (TCR)alpha beta+, as well as showing strong surface expression of HLA-DR, CD25, LFA-1 and ICAM-1. The generated CD4+ T cells secreted interferon (IFN)-gamma, tumour necrosis factor (TNF)-alpha, TNF-beta, and interleukin (IL)-6 (but not IL-4), and exhibited a high level of cytolytic activity against several tumour cell lines. The cytolytic activity of these T cells for Daudi cells was inhibited by preincubation with an anti-intercellular adhesion molecule (ICAM)-1 antibody, but not by preincubation with anti-TCR alpha beta, anti-CD2, or anti-LFA-1 antibodies. Pretreatment with anti-ICAM-1 antibody inhibited T-cell cytolytic activity, but not conjugation with target cells. In addition, incubation with immobilised anti-ICAM-1 enhanced the secretion of IFN-gamma by T cells. We conclude that ICAM-1 expressed on the effector cytotoxic CD4+ T lymphocytes delivers regulatory signals that enhance IFN-gamma secretion.

Adjuvants, Immunologic↗
PubMed1996

Investigation of the oxytocin receptor expression in human breast cancer tissue using newly established monoclonal antibodies.

The expression of the oxytocin (OT) receptor (OTR) in breast cancer was studied using newly established anti-OTR monoclonal antibodies. Immunoblotting indicated that the antibody 2F8 recognized a 70K OTR in the pregnant myometrium and breast cancer tissue. Among 57 breast cancer patients, we detected OTR immunoreactivity in 52 (91.2%) by immunohistochemistry using 2F8. Using another monoclonal antibody for different receptor domains, 1-2, the staining profile was identical in all positive samples. Of 52 OTR-positive samples, 28 were diffusely positive (> 80% of cancer cells were stained), and 24 were partially positive (< 80% cells were stained). The ratio of estrogen receptor-positive samples was slightly higher among those that were diffusely positive, but there was no apparent relationship between OTR expression and other clinical parameters. We also confirmed the expression of the OTR in positively stained samples by means of Northern blotting and RT-PCR at the transcription level. The OTR messenger RNA and RT-PCR product were the same size as those in the pregnant myometrium. We also determined the expression of the OTR using flow cytometry in four breast cancer cell lines (MCF-7, MDA-MB-231, MDA-MB-361, and MDA-MB-468). However, OT had no significant effect on their growth during a short period (7 days) of culture. These findings indicated that the OTR is expressed in breast cancer derived not from the myoepithelium but from the glandular or ductal epithelium; however, the biological function of OT in breast cancer remains to be determined.

Adult↗
PubMed1995

Subcellular localization of the APC protein: immunoelectron microscopic study of the association of the APC protein with catenin.

Mutations in the APC gene are linked to the development of sporadic colorectal tumors as well as to familial adenomatous polyposis. Recently, the APC protein was reported to associated with catenins, proteins that bind to the cell adhesion molecule E-cadherin. In the present study, we examined the distribution and localization of the APC protein and alpha -catenin in the normal mouse intestine by light and immunoelectron microscopy using specific antibodies. The APC protein was found to be localized in microvilli and in the apical and lateral cytoplasm of the epithelial cells, whereas alpha-catenin was detected only in the lateral cytoplasm. Double-labeling immunoelectron microscopy showed colocalization of the APC protein with alpha-catenin in the lateral cytoplasm, especially along the lateral plasma membrane, although a certain portion of the APC protein in this region was distributed independently of alpha-catenin. These results suggest that a portion of the APC protein localized in the lateral cytoplasm of intestinal epithelial cells functions in cooperation with catenins, whereas the APC protein in microvilli and in the apical cytoplasm has other functions independent of catenins.

Adenomatous Polyposis Coli Protein↗
PubMed1995

Coexpression of cdk2/cdc2 and retinoblastoma gene products in colorectal cancer.

The retinoblastoma gene (Rb gene) is a tumour-suppressor gene and its product (pRB) is known to act as a negative regulator of the cell cycle. Although lack of pRB expression resulting from gene alterations is considered to be responsible for the genesis of several human malignancies, increased expression of pRB has been demonstrated in a majority of colorectal cancer cases. In the present study, we investigated the expression of pRB as well as that of its related kinases, cdk2 and cdc2, in colorectal cancer, since these kinases have been reported to phosphorylate and inactivate pRB. Western blot analysis revealed that colorectal cancer expressed higher levels of cdk2 and cdc2 than did normal mucosa and that the ratio of the hyperphosphorylated form of pRB was higher in colorectal cancer. Furthermore, immunohistochemical studies showed that cdk2/cdc2 was expressed exclusively in the cancer cells positive for pRB. These results suggest that an increase in the expression of cdk2/cdc2 in colorectal cancer may have prevented pRB from braking the cell cycle through phosphorylation.

Blotting, Western↗
PubMed1994

Germ-line and somatic mutations of the APC gene in patients with Turcot syndrome and analysis of APC mutations in brain tumors.

The Turcot syndrome (TS) is a rare, probably autosomal recessive, disorder characterized by development of primary neuroepithelial tumors of the central nervous system (CNS) and numerous adenomatous colorectal polyps. To examine the possible involvement of mutations of the APC gene, which is responsible for familial adenomatous polyposis (FAP), in Turcot syndrome, we examined DNAs from TS patients for alterations in this gene by means of ribonuclease protection analysis. Germ-line APC mutations were detected in each of three unrelated cases of TS, and additional (somatic) mutations were observed in colonic adenomas that had developed in one of these patients. However, no somatic mutations in APC were found among 91 neuroepithelial tumors (medulloblastoma, glioblastoma, astrocytoma, and oligodendroglioma), whether sporadic or associated with TS. These results suggest that the APC gene is associated with pathogenesis of one feature of TS, but that at least one other gene is responsible for the genesis of neuroepithelial tumors in the CNS.

Adenomatous Polyposis Coli↗
PubMed1994

Immunohistochemical study of p53 in gastric carcinoma.

The authors had previously reported that the accumulation of p53 is clearly demonstrable in microwave-fixed, paraffin-embedded sections of colorectal carcinomas. In the present paper, the authors performed an immunohistochemical study of p53 in gastric carcinomas prepared by microwave irradiation. Using a monoclonal antibody (PAb1801), nuclear p53 was detected in 22 of 45 (49%) cases of gastric carcinoma, and no staining for p53 was demonstrated in the adjacent normal epithelium, including areas of intestinal metaplasia. The incidence of p53 accumulation was not affected by clinical features, such as tumor stage, depth of invasion, and metastasis; however, positivity for p53 was significantly higher in intestinal-type carcinomas (56%) than in diffuse-type (27%) carcinomas. Furthermore, a large number of carcinoma cells expressed p53 in the intestinal-type, but only a few cells were positive for p53 in the diffuse-type. The results of the present immunohistochemical study of p53 accumulation in gastric carcinomas suggest that gene alterations of p53 are not rare in gastric carcinomas and may participate in the carcinogenesis of intestinal-type carcinomas of the stomach.

Gastric Mucosa↗
PubMed1994

Immunohistochemical analysis of p53 in gynecologic tumors.

Immunohistochemical staining for the p53 protein was performed in microwave-fixed, paraffin-embedded sections of normal, premalignant and malignant tissues of the female genital tract using a monoclonal antibody, PAb 1801. No staining was detected in normal and premalignant tissues, whereas nuclear staining of cancer cells was observed in 12 (22%) of 55 cervical squamous cell carcinomas, 4 (25%) of 16 cervical adenocarcinomas, 37 (42%) of 88 endometrial carcinomas, 23 (38%) of 60 ovarian adenocarcinomas, and 6 (100%) of 6 squamous cell carcinomas arising in dermoid cysts. Of interest, 1 of 7 endometrial cancers with concomitant atypical hyperplasia showed weak nuclear staining in a few atypical hyperplastic glands in addition to the cancerous lesions. Although staining was associated with cancers having a high histologic grade and serous papillary adenocarcinomas of the endometrium, it did not correlate with invasion, metastasis, or clinical stage. Comparison of the staining patterns with molecular analysis of mutations in the p53 gene showed the expected correlation of nuclear staining with missense mutations but not with nonsense mutations, which consistuted one third of all mutations found in this series. In addition, cytoplasmic staining did not predict mutation.

Cervix Uteri↗
PubMed1994

Pancreas cancer-associated antigen (PCAA) in medullary thyroid carcinoma.

Pancreas cancer-associated antigen (PCAA), primarily isolated from the ascites of pancreatic cancer (PC) patients, is strongly positive in PC, colon cancer and normal colonic mucosa. In immunohistochemical staining of tumor tissues with antibodies, medullary thyroid carcinoma (MTC) was no less strongly positive for PCAA than PC, and we studied its details. Antibodies to PCAA, calcitonin and CEA were used in the immunostaining of normal thyroid tissues and thyroid tissues from patients with adenomas, MTC, papillary carcinomas, and follicular carcinomas. The PCAA from the liver metastases of MTC was studied for molecular weight and antigenicity in comparison with the PCAA from the ascites of PC patients. Serum levels of PCAA were determined in MTC patients. Of 11 patients with MTC, PCAA, calcitonin and CEA were studied immunohistologically and positive in 10, 11 and 10 patients, respectively. The PCAA from the metastases had a molecular weight of about 700,000, and was immunochemically identical to that from the ascites of PC patients. Serum levels of PCAA were elevated in 4 of 6 MTC patients. The thyroid tissues from the MTC patients, familial or non-familial, were as strongly positive for PCAA as for calcitonin and CEA. It was antigenically identical to that of PC origin, and positive in the serum of MTC patients.

Adenocarcinoma, Follicular↗
PubMed1993

Significance of p53 expression as a prognostic factor in oesophageal squamous cell carcinoma.

The tumour suppressor gene product p53 is believed to play an important role in the progression of human malignant tumours through mutation and over-expression. Using a microwave oven heating method, we have detected over-expression of p53 in buffered-formalin fixed, paraffin-embedded sections of oesophageal carcinomas immunohistochemically and examined the relationship between the p53 over-expression and postoperative survival. Employing a monoclonal antibody (pAb1801), nuclear p53 was detected in 56 of 105 (53%) tumour specimens. Homogeneous, heterogeneous, and focal immunostaining patterns were noted. No immunostaining was found in adjacent benign tissues. The results in buffered-formalin fixed sections were similar to those in the frozen sections. The cumulative survival rate of patients with p53 expression was significantly lower than that of the patients without expression (P < 0.05), even though there were no significant differences between the clinicopathological features of the two groups. The results indicate that the nuclear accumulation of p53 might be an independent prognostic factor in patients with oesophageal squamous cell carcinomas.

Adult↗
PubMed1993

Augmentation of antitumor immunity in regional lymph nodes by local immunotherapy.

We have previously reported that the antitumor effect of OK-432, a streptococcal preparation, is markedly augmented when injected intratumorally together with fibrinogen (OK-432/fbg) [1]. In order to elucidate the effects of this immunotherapy on regional lymph nodes (RLN), we carried out both morphological and functional analyses of the RLN from colonic cancer patients treated with OK-432/fbg. Computer-aided morphometry revealed that the maximal cross-sectional areas and the broadest diameters of the RLN were significantly greater (p < 0.01) in patients who had undergone local immunotherapy than in patients who had not. The component structures of RLN, such as sinus, follicle and paracortex, were all enlarged in the OK-432/fbg-treated patients, and necrosis of metastatic tumors was observed. RLN lymphocytes recovered from OK-432/fbg treated patients showed elevated reactivity to phytohemagglutinin (PHA) and the stimulation index was clearly higher than that of control patients. Flow cytometric analysis revealed a predominance of T-cells, especially CD4 subsets, and higher positivity for both CD25 and HLA-DR. Furthermore, RLN lymphocytes killed more effectively K562 and Daudi cells in the patients who had had immunotherapy. These results suggest that the effect of local immunotherapy with OK-432/fbg is not restricted to the site of injection but extends to the lymph nodes, and contributes to tumor regression through the augmentation of cellular immunity.

Aged↗
PubMed1993

The effect of local immunotherapy for breast cancer using a mixture of OK-432 and fibrinogen supplemented with activated macrophages.

OK-432 is an immunomodulatory agent prepared from a strain of Streptococcus pyogenes. We have previously reported that intratumoral injection of a mixture of OK-432 and fibrinogen (hereinafter referred to as OK/fbg) is very effective in the local immunotherapy for colorectal cancer. However, we found that the intratumoral injection of OK/fbg into tumor tissues of breast cancers did not always induce a strong antitumor effect. With conventional OK/fbg treatment, tumor necrosis observed in breast cancer tumors was significantly less than that in colorectal cancer tumors; the formation of fibrin meshwork and macrophage infiltration, in particular, were poor. In this study, the OK/fbg mixture was supplemented with activated macrophages for local immunotherapy of breast cancers. Macrophages were prepared from peripheral blood of breast cancer patients and activated with 0.05 mg/ml of OK-432. Between 2-7 days before operation, a single intratumoral injection of the above mixtures was done. The addition of activated macrophages to the OK/fbg mixture resulted in marked degrees of fibrin meshwork formation, macrophage infiltration and cancer cell necrosis. These findings suggest that the recruitment of macrophages in tumor stroma and their activation are necessary for sufficient induction of antitumor immunity, and supplementation of activated macrophages at the site of immune reaction may be an alternative method for reinforcement of the antitumor effect of local immunotherapy.

Breast Neoplasms↗
PubMed1993

A cancer-reactive human monoclonal antibody derived from a colonic cancer patient treated with local immunotherapy.

A human monoclonal antibody, YJ-37 (IgM) was generated through the fusion of human B lymphoblastoid cell line HO-323 with the regional lymph node lymphocytes from a colonic cancer patient who was treated with a local immunotherapy. This antibody was purified and conjugated with biotin, after which direct immunohistochemical staining was performed. The results revealed that YJ-37 selectively reacted with colonic cancer (7/19), gastric cancer (3/6), endometrial cancer (1/2) and colonic adenoma (7/13), but not with normal epithelia. Membrane immunofluorescence and FACS analysis also showed that YJ-37 bound to tumor cell surfaces. Furthermore, the chemical structure of the antigen defined by YJ-37 was analyzed by means of thin-layer chromatography immunostaining and ELISA. The results indicated that YJ-37 reacted with sialylated lacto-series carbohydrate chains, which have been reported to accumulate in cancer cells.

Adenoma↗
PubMed1993

Preparation of a human monoclonal antibody derived from cervical lymph nodes of a patient with anaplastic carcinoma of the thyroid.

An anaplastic carcinoma cell line, KOA-2, was established from a 59-year-old patient with anaplastic carcinoma of the thyroid who died only one month after diagnosis in association with rapid progress of her disease. A human monoclonal antibody, 3C5 (IgM), was generated by a fusion of human B lymphoblastoid cell line, HO-323, with lymphocytes from cervical lymph nodes of the patient, who had been treated with local immunotherapy. Immunocytostaining demonstrated that 3C5 reacted with KOA-2, and membrane immunofluorescence demonstrated that 3C5 reacted with cell membranes. Immunohistochemical staining studies demonstrated selective reaction of 3C5 with three malignant tumors, anaplastic carcinoma of the thyroid (2/4 sampled tested), papillary carcinoma of the thyroid (8/12), and breast cancer (2/6), but no reaction with specimens of benign thyroid disease or normal thyroid gland was demonstrated. The monoclonal antibody and the cell line established from one patient have potential use in the analysis of carcinogenesis and applications to therapy of anaplastic carcinoma of the thyroid.

Antibodies, Monoclonal↗
PubMed1992

Use of fibrinogen to enhance the antitumor effect of OK-432. A new approach to immunotherapy for colorectal carcinoma.

OK-432 (5 KE), an immunomodulatory agent prepared from an attenuated strain of Streptococcus pyogenes, was dissolved in 1 ml of aprotinin (1000 KIE) and mixed with 80 mg of fibrinogen containing Factor XIII. A single intratumoral injection of the mixture was performed preoperatively under endoscopy in 20 patients with colorectal carcinoma. Postoperative histopathologic examinations revealed the formation of fibrin fibers at the site of injection and marked infiltration of inflammatory cells into the tumor stroma on the day after injection; the formation of granulomas containing many giant cells after 4 to 7 days; and extensive regression of tumor tissue after 14 days. This study suggests that the high concentration of exogenous fibrinogen gelatinized enough to trap OK-432 in tumor stroma and that OK-432 induced granulomatous hypersensitivity to degenerate tumor stroma, thereby causing regression of the tumors.

Adenocarcinoma↗