Biomedical subjects

Hideaki Ueno

Publications and source records attributed to Hideaki Ueno.

6 recordsLinked to original sources

PubMed2006

Maturation of dendritic cells and T-cell responses in sentinel lymph nodes from patients with breast carcinoma.

BACKGROUND: Methods for identifying sentinel lymph nodes (SNs) and their clinical significance have been established. Recent advances in molecular immunology have enabled the analysis of precise immune responses. The objective of the current study was to clarify the dendritic cell (DC) maturation, T-helper type 1 (Th-1) and Th-2 responses, and regulatory T-cell responses of SNs in patients with breast carcinoma. METHODS: SNs and non-SNs were identified by radioguided and blue dye-guided methods in 70 consecutive patients with clinically lymph node negative (N0) breast carcinoma. Lymphocytes were collected from SNs and non-SNs and were subjected to flow cytometric analysis (FCM) using antibodies of CD83-fluorescein isothiocyanate (FITC), CD80-phycoerythrin (PE), CD86-PE, CD40-PE, human leukemic D-related antigen (HLA-DR)-FITC, CD4-FITC, and CD25-PE. Total RNA was extracted from SNs and non-SNs, and the expression of CD83, interleukin 12p40 (IL-12p40), interferon gamma (IFN-gamma), IL-4, IL-10, and Foxp3 was evaluated by using quantitative real-time reverse transcriptase-polymerase chain reaction (RT-PCR) analysis. The immunologic status of SNs was analyzed further with regard to micrometastases, which were identified as negative microscopically but positive according to an RT-PCR analysis that was specific for mammaglobin. RESULTS: SNs were detectable in 70 of 71 consecutive patients (98.6%) with clinically N0 breast carcinoma. Fourteen of 70 patients (20.0%) had positive metastasis in SNs. When SNs were compared with non-SNs in 56 metastasis-negative patients, FCM revealed that HLA-DR-positive, CD80-positive, CD86-positive, and CD40-positive cell populations were decreased significantly in SNs. RT-PCR analysis demonstrated that, among 44 patients with metastasis-negative SNs, the expression levels of CD83 and IFN-gamma mRNA were significantly lower in SNs compared with non-SNs. Immunologic parameters also were compared between 44 metastasis-negative SNs and 14 metastasis-positive SNs. The metastasis-positive SNs demonstrated significantly higher expression of CD83, IL-12p40, IFN-gamma, IL-10, and Foxp3 mRNA than the metastasis-negative SNs. Correction of micrometastasis detected by mammaglobin enhanced these differences consistently. CONCLUSIONS: In patients with breast carcinoma, cellular immune responses, from DC maturation to Th-1 responses, appeared to be less active in SNs compared with non-SNs before metastasis developed. Once metastasis was established in SNs, DC maturation was triggered and was followed by the up-regulation of Th-1 responses, which may reflect antigen-specific immune responses in SNs. Unlike DC maturation and Th-1 responses after metastasis in SNs, up-regulation of Th-2 and regulatory T-cell responses developed in parallel.

Adult↗

PubMed2005

Multimarker quantitative real-time PCR detection of circulating melanoma cells in peripheral blood: relation to disease stage in melanoma patients.

BACKGROUND: Detection of melanoma cells in circulation may be important in assessing tumor progression. The objective of this study was to develop a specific, reliable multimarker quantitative real-time reverse transcription-PCR (qRT) assay for detecting melanoma cells in patients' blood. METHODS: We developed qRT assays for the mRNA of four melanoma-associated markers: MART-1, GalNAc-T, PAX-3, and MAGE-A3. In optimization studies, we tested 17 melanoma cell lines and 49 peripheral blood leukocyte (PBL) samples from volunteers. We performed RNA and melanoma cell dilution studies to assess the detection limits and imprecision of the assays. We measured the mRNAs in blood specimens from 94 melanoma patients [American Joint Committee on Cancer (AJCC) stage I, n = 20; II, n = 20; III, n = 32; IV, n = 22]. RESULTS: All markers were frequently detected in melanoma cell lines, whereas none of the markers was detected in PBLs from volunteers. The qRT assay could detect 1 melanoma cell in 10(7) PBLs in the melanoma cell-dilution studies. Markers were detected in 15%, 30%, 75%, and 86% of melanoma patients with AJCC stage I, II, III, and IV disease, respectively. The number of positive markers and AJCC stage were significantly correlated (Spearman correlation coefficient = 0.58; P < 0.0001). CONCLUSIONS: Multimarker qRT can detect circulating melanoma cells in blood. Measurement of the studied molecular markers in blood may be useful in detection of metastasis and monitoring treatment response of melanoma patients.

Antigens, Neoplasm↗

PubMed2003

Future prospects of personalized chemotherapy in gastric cancer patients: results of a prospective randomized pilot study.

In the present study, in order to evaluate the feasibility of personalized chemotherapy, a prospective randomized pilot study was performed in 30 advanced or recurrent gastric cancer patients. As we have demonstrated previously, the expressions of mRNA from tumor biopsy samples for seven molecular markers, i.e., dihydropyrimidine dehydrogenase (DPD), glutathione S-transferase (GST)-pi, beta-tubulin (tub), O6-methylguanine-DNA methyltransferase (MGMT), multiple drug-resistant protein (MRP)-1, NADPH/quinone oxidoreductase (NQO)-1, and cytochrome p450 (P450), were examined by reverse transcription-polymerase chain reaction (RT-PCR) analysis and therapy was recommended in a flow chart that depended on the level of expression of these predictive molecular markers. We chose 12 therapeutic plans, including best supportive care (BSC). We treated 15 patients according to the gene expression profiles, and the remaining 15 patients (controls) were treated without recommended regimens, and the therapy was continued after the expression profiles were checked. Interestingly, 11 of 26 lesions (42.3%) responded after treatment given according to gene expression analysis; however, no clinical response was detected in the control group. The prediction of the response, including resistance, was successful in 75.9% by the gene expression profiles. Moreover, the survival of the patients with the recommended treatment was better than that of patients without a recommended protocol. These results indicate that personalized treatment may be beneficial for gastric cancer chemotherapy and further randomized trials should be carried out in the future.

Adult↗

PubMed2003

[A new tracer for sentinel node detection in esophageal and lung cancer--fluorescent microspheres].

Tracers commonly used for sentinel node detection are blue dye and radioisotope (RI)-labeled colloids. In esophageal cancer or lung cancer, it is impossible to detect the sentinel node among anthracotic mediastinal nodes with blue dye. RI-labeled colloids must be treated in a special facility under complicated regulations. To overcome these problems, we used fluorescent microspheres as a tracer in animals and detected fluorescence-positive nodes using the ultraviolet light irradiation method. Systemic and local injection of fluorescent microspheres revealed no side effects in rats. Submucosal and subadventitial injection of fluorescent microspheres to the esophagus of pigs stained the lymphatic pathway and sentinel nodes under ultraviolet light irradiation. This sentinel node detection system is considered to be useful for sentinel node detection especially for mediastinal nodes that show anthracotic pigmentation. It should be noted that this non-RI method is acceptable at any general medical facility.

Animals↗

PubMedDate not supplied

Quantitative detection of carcinoembryonic antigen messenger RNA in the peritoneal cavity of gastric cancer patients by real-time quantitative reverse transcription polymerase chain reaction.

BACKGROUND: Peritoneal dissemination is the most frequent cause of recurrence in gastric cancer patients after gastrectomy. We have previously demonstrated the prognostic factors in gastric cancer including growth factors and their receptors and intra-peritoneal CEA (1,2). MATERIALS AND METHODS: In the present study, to elucidate whether CEA mRNA in peritoneal lavage is a good marker for the prediction of peritoneal recurrence of gastric cancer patients, quantitative real-time RT-PCR with primers specific for CEA was performed by peritoneal lavage of 124 gastric cancer patients collected at the time of operation. RESULTS: CEA crossing-point values (CEA-CP) were significantly correlated with the depth of tumor invasion and the values were cut-off at 27.11, which was the minimum value of mucosal cancer patients. The CEA-CP was significantly correlated with lymph node metastasis, lymphatic invasion, venous invasion, stages and overall survivals. In prediction of peritoneal recurrence by CEA-CP, sensitivity and specificity were 76.9% and 67.7%, respectively. Cytology predicted peritoneal recurrence with 23.1% sensitivity and 98.1% specificity. CONCLUSION: Quantitative RT-PCR of CEA mRNA of peritoneal lavage is a useful factor for prognosis and predicting peritoneal recurrence.

Biomarkers, Tumor↗

PubMedDate not supplied

Experimental study on fluorescent microspheres as a tracer for sentinel node detection.

BACKGROUND: Common tracers for sentinel node navigation surgery are blue dye and technetium-99m-labelled colloids. However, in most esophageal or lung cancer patients, it is impossible to detect the sentinel node among mediastinal nodes by blue dye because of the anthracotic pigmentation of mediastinal nodes. The use of technetium-99m-labelled colloids requires a special facility, while a large hot-spot at the injection site prevents detection of the sentinel node around the primary lesion. To overcome these problems, we investigated the use of fluorescent microspheres (0.1-20 microm in diameter) as tracers in animals and detected fluorescence-positive nodes by a simple ultraviolet light irradiation method. MATERIALS AND METHODS: Two milliliters of fluorescent microspheres 0.1-20 microm in diameter diluted to 2.5% weight per volume was injected via the tail vein of 30 rats; systemic side-effects were examined. One milliliter of 1.0 microm-diameter microspheres dilution was injected on the backs of 30 rats; local side-effects were examined. A microsphere dilution (0.2 ml, 1.0 microm-diameter microspheres) was injected into the footpad of 18 rats; the lymphatic pathway and drainage were examined. Five milliliters of 1.0 microm-diameter fluorescent microspheres was injected endoscopically into the submucosa of the esophagus, stomach and small and large bowels of 6 domestic pigs, and 5 ml was injected into the subadventitia of the esophagus or subserosa of the stomach, and small and large bowels. Fluorescence-positive lymph ducts or nodes were carefully observed under ultraviolet light irradiation. RESULTS: No systemic side-effects were observed in rats. Only mild edema and a mild inflammatory reaction were observed on the backs of rats. Fluorescent microspheres 0.1, 0.5 and 1.0 microm in diameter were detected in lymph ducts or nodes of pigs within 1 hour after injection. CONCLUSION: Sentinel node navigation surgery with the use of fluorescent microspheres might be feasible and advantageous for patients with esophageal or lung cancer, especially in the mediastinum.

Animals↗