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1.
Acta Med Okayama ; 76(5): 541-545, 2022 Oct.
Article in English | MEDLINE | ID: mdl-36352801

ABSTRACT

The purpose of this study was to conduct a self-reported questionnaire survey of work-related musculoskeletal disorders (WMSDs) among Japanese radiological technologists (RTs) and to report on the relationship between wearing a lead apron and WMSDs. Between February and April of 2021, RTs in Okayama Prefecture, Japan, were surveyed by mail and through a website. Information on individual characteristics, physical factors at work, and the presence of WMSDs were collected. All participants were also asked whether they frequently wore lead aprons. A multiple logistic regression analysis was used to assess the relationship between wearing a lead apron and WMSDs. The model was adjusted for age, sex, body mass index (BMI), and working hours. Of the 123 participants, 67 (54.5%) had WMSDs. Multiple logistic regression analysis revealed that WMSDs were significantly associated with wearing a lead apron. Compared to the "Never wear" group, the odds ratios for the "Always/Frequently wear" and "Sometimes/Rarely wear" groups were 7.87 (95% confidence interval [CI]=1.28-48.46; p=0.026) and 7.80 (95% CI=1.43-42.44; p=0.017), respectively. Our analysis suggests that wearing a lead apron is associated with WMSDs, and thus design modifications in lead aprons may improve the occupational health management of RTs.


Subject(s)
Musculoskeletal Diseases , Occupational Diseases , Humans , Japan/epidemiology , Occupational Diseases/epidemiology , Occupational Diseases/etiology , Musculoskeletal Diseases/epidemiology , Musculoskeletal Diseases/etiology , Surveys and Questionnaires , Risk Factors , Prevalence
2.
Acta Med Okayama ; 71(3): 219-226, 2017 Jun.
Article in English | MEDLINE | ID: mdl-28655941

ABSTRACT

 Anticancer drugs induce cell-cycle arrest and apoptosis not only in tumor cells, but also in immune cells. However, many preclinical and clinical findings show that some chemotherapeutic agents can improve the antitumor efficacy of immunotherapy. We immunohistochemically analyzed the degree of immune cell infiltration and the relevance of programmed cell death 1 ligand-1 (PD-L1) expression in surgically resected oral squamous cell carcinoma (OSCC) specimens from patients who had undergone pretreatment with certain chemotherapies and other patients without pretreatment. We divided the patients into the group of neoadjuvant chemotherapy (NAC) patients (n=8) and the nNAC (without NAC) patient group (n=10). We observed that NAC induced infiltrations of CD4, CD8 T cells and CD56 NK cells into the tumor microenvironment. Decreased numbers of Tregs and PD-1-positive cells were observed in the NAC group. No significant difference was observed in the degree of immune-cell infiltration between the patient groups except for CD56 NK cells in the stroma and PD-1 cells in cancer nests. Eighty percent of the nNAC specimens showed intermediate-to-strong PD-L1 protein expression, whereas 75% of the NAC specimens showed down-regulation of the PD-L1 protein, indicating the effectiveness of the chemotherapeutic treatment before surgery.


Subject(s)
Antineoplastic Agents/therapeutic use , B7-H1 Antigen/metabolism , Carcinoma, Squamous Cell/immunology , Mouth Neoplasms/immunology , Neoadjuvant Therapy , Tumor Microenvironment/drug effects , Adult , Aged , Aged, 80 and over , Carcinoma, Squamous Cell/drug therapy , Carcinoma, Squamous Cell/pathology , Carcinoma, Squamous Cell/surgery , Case-Control Studies , Drug Combinations , Female , Humans , Male , Middle Aged , Mouth Neoplasms/drug therapy , Mouth Neoplasms/pathology , Mouth Neoplasms/surgery , Oxonic Acid/therapeutic use , Retrospective Studies , Tegafur/therapeutic use , Tumor Microenvironment/immunology
3.
J Radioanal Nucl Chem ; 307: 1279-1285, 2016.
Article in English | MEDLINE | ID: mdl-26834308

ABSTRACT

Hydroxyapatite (HAP) is a main mineral constituent of bone and tooth and has an outstanding biocompatibility. HAP is a possible sorbent for heavy metals in wastewater due to its high adsorption capacity and low water solubility. We developed a removal system of 90Sr from aqueous solution by HAP column procedure. More than 90 % of 90Sr was adsorbed and removed from the 90Sr containing solution. Divalent cations, Ca2+, had little effect on the removal of 90Sr up to a concentration of 1 mmol L-1. This clearly indicates that the HAP column technique is advantageous with respect to the capacity to adsorb 90Sr from water present in the environment.

4.
J Plant Res ; 127(1): 11-22, 2014.
Article in English | MEDLINE | ID: mdl-24346655

ABSTRACT

One year after the deposition of radionuclides from the Fukushima 1 Nuclear Power Plant (A formal name is Fukushima Daiichi Nuclear Power Station) in March 2011, radiocesium (¹³4Cs, ¹³7Cs) concentrations ([Cs]) were comprehensively investigated in the wild plants of 99 species most of which were annual or summer green perennial herbs and started to grow from April 2012 at the heavily contaminated fields of paddy (three study sites) and upland (one study site) in Fukushima Prefecture. The survey was conducted three times (April, July and October) in the year. In each site, soils (soil cores of 5-cm depth) and plants (aerial shoots) were collected for determination of [Cs] on a dry weight basis, and then the transfer factor (TF) of radiocesium from soil to plant ([Cs]plant/[Cs]soil) was estimated in each species. The [Cs] values of both soils and plants largely varied. However, some species exhibited relatively high TF values (more than 0.4) (e.g., Athyrium yokoscense, Dryopteris tokyoensis, and Cyperus brevifolius), while others exhibited almost negligible values (less than 0.01) (e.g., Salix miyabeana, Humulus scandens, and Elymus tsukushiensis). In addition, judging from the 11 species grown in both paddy and upland fields, TF values were generally higher in the paddy fields. The estimation of phytoextraction efficiency of soil radiocesium by weed communities in the paddy fields suggests that the weed community is not a practical candidate for phytoremediation technique.


Subject(s)
Cesium Radioisotopes/metabolism , Fukushima Nuclear Accident , Plants/metabolism , Soil/chemistry , Biodegradation, Environmental , Cesium Radioisotopes/analysis , Japan , Nuclear Power Plants , Plants/chemistry , Radiation Monitoring , Soil Pollutants, Radioactive/analysis , Soil Pollutants, Radioactive/metabolism , Species Specificity
5.
J Radioanal Nucl Chem ; 298(1): 295-300, 2013.
Article in English | MEDLINE | ID: mdl-26224937

ABSTRACT

Ion exchange is a simple and efficient method for separating no-carrier-added 64Cu from an irradiated Ni target. We developed a semi-automated two-round 64Cu separation system equipped with a strong-base anion exchange resin column. We first verified the efficiency of the system using a non-radioactive substitute consisting of 25 mg of Ni and 127 ng of Cu, and confirmed that Cu was completely eluted at the second round of the separation step. After the bombardment, separation of 64Cu from the Ni target was achieved with high radiochemical purity. 64Cu produced and separated in this study had an extremely low level of Ni impurity. It could be used for labeling monoclonal antibodies for antibody positron emission tomography imaging and synthesizing radiopharmaceuticals.

6.
BMC Cancer ; 12: 520, 2012 Nov 14.
Article in English | MEDLINE | ID: mdl-23151147

ABSTRACT

BACKGROUND: Identification of new cancer antigens is necessary for the efficient diagnosis and immunotherapy. A variety of tumor antigens have been identified by several methodologies. Among those antigens, cancer/testis (CT) antigens have became promising targets. METHODS: The serological identification of antigens by the recombinant expression cloning (SEREX) methodology has been successfully used for the identification of cancer/testis (CT) antigens. We performed the SEREX analysis of colon cancer. RESULTS: We isolated a total of 60 positive cDNA clones comprising 38 different genes. They included 2 genes with testis-specific expression profiles in the UniGene database, such as TEKT5 and a CT-like gene, A kinase anchoring protein 3 (AKAP3). Quantitative real-time RT-PCR analysis showed that the expression of TEKT5 was restricted to the testis in normal adult tissues. In malignant tissues, TEKT5 was aberrantly expressed in a variety of cancers, including colon cancer. A serological survey of 101 cancer patients with different cancers by ELISA revealed antibodies to TEKT5 in 13 patients, including colon cancer. None of the 16 healthy donor serum samples were reactive in the same test. CONCLUSION: We identified candidate new CT antigen of colon cancer, TEKT5. The findings indicate that TEKT5 is immunogenic in humans, and suggest its potential use as diagnostic as well as an immunotherapeutic reagent for cancer patients.


Subject(s)
Antigens, Neoplasm/blood , Antigens, Neoplasm/immunology , Microtubule Proteins/blood , Microtubule Proteins/immunology , Neoplasms/immunology , Testis/immunology , Antibody Formation , Antigens, Neoplasm/genetics , Antigens, Neoplasm/metabolism , Cloning, Molecular/methods , DNA, Complementary/genetics , Humans , Male , Microtubule Proteins/genetics , Microtubule Proteins/metabolism , Neoplasms/blood , Neoplasms/genetics , Neoplasms/metabolism , Serologic Tests/methods , Transcriptome/genetics , Transcriptome/immunology
7.
Acta Med Okayama ; 65(2): 91-5, 2011 Apr.
Article in English | MEDLINE | ID: mdl-21519366

ABSTRACT

No research has been conducted on the radiation influence of tobacco on the alimentary system, although there have been some previous works on the respiratory system. In this study, the radioactive concentrations of 210Pb and 40K in a cigarette sample were first measured. The transfer factors of the nuclides from tobacco into smoke and solution (saliva and/or alcohol) were then examined. Moreover, the radiation doses from smoke inhalation were also evaluated. The radioactive concentrations of 210Pb and 40K in the cigarette tobacco were 0.01 and 0.3 Bq/cigarette. Since this 210Pb activity and the 210Po activity previously reported for the same sample were comparable, it can be concluded that there was a radioactive equilibrium between the 2 nuclides. The observed transfer factor of 210Pb (12%) into smoke was almost the same as that of 40K (15%), whereas the reported value for 210Po (60%) was significantly higher. The radiation doses due to inhalation of cigarette smoke varied from organ to organ, depending on the organotropic properties of the nuclide. For example, the kidneys, respiratory tract, and spleen showed relatively high doses from 210Pb and 210Po. The leaching rates indicated an inconsistent tendency related to solution types. This result could suggest that alcohol drinking, which is common in smokers, does not especially enhance the leaching characteristics.


Subject(s)
Lead Radioisotopes/analysis , Nicotiana/chemistry , Potassium Radioisotopes/analysis , Smoking , Alcohol Drinking , Humans , Inhalation Exposure , Monte Carlo Method
8.
Cancer Lett ; 301(1): 57-62, 2011 Feb 01.
Article in English | MEDLINE | ID: mdl-21094583

ABSTRACT

Serological analysis of a recombinant cDNA expression library (SEREX) derived from two lung adenocarcinoma cancer cell lines using autologous sera led to the isolation of 41 positive cDNA clones comprising 28 different antigens. They coded for a variety of nuclear and cytoplasmic proteins. Among the antigens, nucleoporin 107 (NUP107) was isolated most frequently (5 of 41 clones). The second most frequently isolated antigen was coded for by C21orf58 (4 of 41 clones). During serological analysis of selected antigens based on their reactivity to sera from normal individuals and lung cancer patients, none of the antigens showed a cancer-restricted recognition pattern. However, five genes including NUP107 showed higher expression when we examined the changes in gene expression in five different adenocarcinoma cell lines, including those used in SEREX, compared with their levels in normal lung tissues by cDNA microarray analysis. On the other hand, the expression levels of five genes including C21orf58 were down regulated in all adenocarcinoma cell lines. This SEREX study combining comprehensive gene expression assays has added to the growing list of lung cancer antigens, which may aid the development of diagnostic and immunotherapeutic reagents for patients with lung cancer.


Subject(s)
Adenocarcinoma/immunology , Antibodies, Neoplasm/immunology , Antigens, Neoplasm/isolation & purification , Gene Library , Lung Neoplasms/immunology , Antigens, Neoplasm/genetics , Antigens, Neoplasm/immunology , Cell Line, Tumor , Humans
9.
Anticancer Res ; 30(12): 4925-35, 2010 Dec.
Article in English | MEDLINE | ID: mdl-21187472

ABSTRACT

Pulmonary surfactant has been used as a carrier to deliver a therapeutic virus to dysfunctional lung cells that reside within an intricate lung structure. To investigate whether pulmonary surfactant enhances the efficacy of intratracheal instillation of a therapeutic virus to target KRAS mutation-bearing lung cancer in vivo, we developed a recombinant adenovirus that induces cell death only in lung cancer cells and injected the adenovirus into a mouse model of KRAS mutation-positive lung cancer intratracheally with and without surfactant. A therapeutic adenovirus that induces cell death only in lung cancer cells was constructed by combining a cancer-specific human telomerase reverse transcriptase (hTERT) promoter fused to CCAAT/enhancer-binding protein alpha (CEBPα) with a modified lung-specific Clara cell-specific 10-kDa protein (CC10) promoter fused to cytotoxic adenovirus type 5 early region 1A (E1A). CEBPα is induced only in cancer cells and activates the CC10 promoter, which in turn induces cytotoxic E1A, and causes cell death only in lung cancer cells in vitro. This adenovirus was intratracheally administered to the model mice (CCSP-rtTA/Tet-op-K-Ras4bG12D bitransgenic mice) in the presence and absence of pulmonary surfactant. Intratracheally administered therapeutic adenovirus with pulmonary surfactant spread to airways, as well as to the alveolar region of the lung, and caused a reduction of lung tumors developed. The therapeutic adenovirus without pulmonary surfactant spread only to airways and was ten-fold less effective in tumor reduction. Here, we demonstrate that pulmonary surfactant is an efficient tool to intratracheally deliver a therapeutic virus to treat KRAS mutation-positive lung cancer in vivo.


Subject(s)
Adenoviridae/physiology , Gene Transfer Techniques , Genes, ras , Lung Neoplasms/genetics , Lung Neoplasms/therapy , Oncolytic Virotherapy/methods , Pulmonary Surfactants/administration & dosage , Adenocarcinoma/genetics , Adenocarcinoma/therapy , Adenocarcinoma/virology , Adenoviridae/genetics , Adenovirus E1A Proteins/genetics , Animals , Breast Neoplasms , Cattle , Cell Line, Tumor , Female , Genetic Therapy/methods , Genetic Vectors/genetics , HeLa Cells , Humans , Lung Neoplasms/virology , Mice , Mice, Inbred ICR , Mice, Transgenic , Mutation , Promoter Regions, Genetic
10.
Biosci Biotechnol Biochem ; 73(6): 1425-8, 2009 Jun.
Article in English | MEDLINE | ID: mdl-19502754

ABSTRACT

The cbnA gene encoding chlorocatechol dioxygenase from the soil bacterium Ralstonia eutropha NH9 under the control of a modified cauliflower mosaic virus 35S promoter was introduced into a hybrid poplar (Populus tremula x P. tremuloides). Integration of the cbnA gene in transgenic poplar was confirmed by PCR and genomic Southern blot analysis. Expression of the cbnA gene was analyzed by Western blot analysis. Transgenic poplar calli efficiently converted 3-chlorocatechol to 2-chloro-cis,cis-muconate.


Subject(s)
Cupriavidus necator/enzymology , Dioxygenases/genetics , Populus/genetics , Blotting, Southern , Blotting, Western , Chromatography, Liquid , Genes, Plant , Plants, Genetically Modified , Polymerase Chain Reaction , Populus/cytology
11.
Int J Cancer ; 124(10): 2347-52, 2009 May 15.
Article in English | MEDLINE | ID: mdl-19165854

ABSTRACT

Cancer/testis (CT) antigens are expressed in normal germ line tissues and various cancers. They are considered promising target molecules for immunotherapy for patients with various cancers. To identify CT antigens, we performed serological identification of antigens by recombinant expression cloning. The humoral immune response of cancer patients against a newly defined antigen was analyzed. A testicular cDNA library was immunoscreened with serum obtained from a gastric adenocarcinoma patient whose primary cancer had regressed once and most liver metastases had disappeared transiently. We isolated 55 positive cDNA clones comprising 23 different genes. They included 4 genes with testis-specific expression profiles in the Unigene database, including coiled-coil domain containing 62 (CCDC62). RT-PCR analysis showed that the expression of 2 splice variants of CCDC62 was restricted to the testis in normal adult tissues. In malignant tissues, CCDC62 variant 2 (CCDC62-2) was aberrantly expressed in a variety of cancers, including stomach cancer. A serological survey of 191 cancer patients with a range of different cancers by ELISA revealed antibodies to CCDC62-2 in 13 patients, including stomach cancer. None of the 41 healthy donor serum samples were reactive in the same test. The serum reaction against CCDC62-2 was confirmed by western blot. CCDC62-2 is a CT antigen that is immunogenic in cancer patients.


Subject(s)
Adenocarcinoma/immunology , Antigens, Neoplasm/immunology , Stomach Neoplasms/immunology , Testis/immunology , Transcription Factors/immunology , Aged , Antigens, Neoplasm/biosynthesis , Antigens, Neoplasm/genetics , Base Sequence , Blotting, Western , DNA Primers , DNA, Complementary , Enzyme-Linked Immunosorbent Assay , Humans , Male , RNA, Messenger/genetics , Recombinant Proteins/biosynthesis , Recombinant Proteins/genetics , Recombinant Proteins/immunology , Reverse Transcriptase Polymerase Chain Reaction , Transcription Factors/biosynthesis , Transcription Factors/genetics
12.
Cancer Sci ; 99(7): 1441-7, 2008 Jul.
Article in English | MEDLINE | ID: mdl-18452555

ABSTRACT

We have previously shown that the RLakt antigen was predominantly recognized by CD8 cytotoxic T lymphocytes (CTL) in RL male 1-bearing or -rejected syngeneic BALB/c mice. CD8 CTL were directed to the octamer pRL1a peptide IPGLPLSL of which recognition was H-2L(d)-restricted. In this study, we identified a CD4 T-cell epitope peptide in the tumor rejection antigen RLakt on BALB/c radiation-leukemia RL male 1. Analyses of the recognition of a bulk CD4 T-cell line using several recombinant RLakt proteins suggested the presence of multiple CD4 T-cell epitopes in the molecule. However, cloning from a bulk CD4 T-cell line resulted in only two clones from 200 wells seeded at three cells per well, and those two CD4 T-cell clones recognized the same epitope peptide in RLakt. The epitope peptide was 14-mer p12-25, AYREETLSIIPGLP, and its recognition was H-2IA(d)-restricted. This sequence overlapped with the CD8 T-cell epitope pRL1a in its N-terminal 5 amino acid residues. The relationship of the epitope to the pRL1a peptide predominantly recognized by CD8 CTL suggests that the 14-mer epitope is predominantly recognized by CD4 T-cells.


Subject(s)
Antigens, Neoplasm/immunology , CD4-Positive T-Lymphocytes/immunology , Epitopes, T-Lymphocyte , Leukemia, Radiation-Induced/immunology , Animals , Cell Line , Female , Histocompatibility Antigens Class II/immunology , Lymphocyte Activation , Male , Mice , Mice, Inbred BALB C , T-Lymphocytes, Regulatory/immunology
13.
Int J Oncol ; 29(4): 903-10, 2006 Oct.
Article in English | MEDLINE | ID: mdl-16964386

ABSTRACT

OY-TES-1 is a novel target that belongs to the family of 'cancer/testis' (CT) antigens. Our goal was to examine the expression and immunogenicity of OY-TES-1 in epithelial ovarian cancer (EOC) to determine its potential as a target for vaccine therapy. OY-TES-1 expression was determined by one-step reverse transcriptase PCR on 100 EOC samples, 5 EOC cell lines, and a panel of normal tissues. Immunohistochemistry (IHC) was performed on the same panel of EOC tissues. Sera from a sub-group of patients were tested for OY-TES-1 antibody by ELISA. Thymus and leukocytes were weakly positive for OY-TES-1 while the remaining 5 normal tissues were negative. Expression of OY-TES-1 by either RT-PCR and/or IHC was demonstrable in 69/100 (69%) tumors. Humoral immunity to OY-TES-1 was demonstrated in 1/10 (10%) serum samples from patients whose tumors expressed the antigen. The median follow-up of the patient population was 34 months. There was no correlation between antigen expression and stage, grade, histology and survival. OY-TES-1 is expressed in 69% of patients with EOC, is absent from normal ovarian tissue, and a proportion of patients show evidence of a specific humoral immune response. These findings make OY-TES-1 an attractive target for antigen-specific immunotherapy in EOC.


Subject(s)
Antibodies, Neoplasm/blood , Carcinoma/metabolism , Carrier Proteins/metabolism , Ovarian Neoplasms/metabolism , Adult , Aged , Aged, 80 and over , Animals , Antibodies, Monoclonal/immunology , Carcinoma/chemistry , Carcinoma/pathology , Carrier Proteins/analysis , Carrier Proteins/genetics , Female , Humans , Immunohistochemistry , Leukocytes/chemistry , Mice , Middle Aged , Ovarian Neoplasms/chemistry , Ovarian Neoplasms/pathology , RNA, Messenger/analysis , RNA, Messenger/metabolism , Thymus Gland/chemistry
14.
J Gastroenterol Hepatol ; 21(8): 1281-5, 2006 Aug.
Article in English | MEDLINE | ID: mdl-16872310

ABSTRACT

BACKGROUND AND AIM: The present study was designed to investigate the expression of and humoral response against NY-ESO-1 in patients with hepatocellular carcinoma and to analyze the relationship between expression of NY-ESO-1 mRNA and clinicopathological features. METHODS: NY-ESO-1 mRNA and protein expression in surgically resected hepatocellular carcinoma specimens, adjacent non-cancerous liver and non-tumor bearing liver were examined by reverse transcription-polymerase chain reaction and immunohistochemical staining using a monoclonal antibody against NY-ESO-1 (ES121), respectively. The antibody response to NY-ESO-1 was examined by enzyme-linked immunosorbent assay using recombinant NY-ESO-1 protein. RESULTS: NY-ESO-1 mRNA was detected in 18 of 41 (43.9%) hepatocellular carcinomas. No NY-ESO-1 mRNA was expressed in 41 paired non-cancerous specimens and 18 specimens histologically diagnosed as liver cirrhosis or chronic hepatitis. Immunohistochemistry revealed heterogeneous expression of NY-ESO-1 protein in three of 18 NY-ESO-1 mRNA-positive hepatocellular carcinomas. None of 23 NY-ESO-1 mRNA-negative hepatocellular carcinomas expressed NY-ESO-1 protein. Antibody against NY-ESO-1 protein was detected in two of 92 patients with hepatocellular carcinoma. Both of these patients had tumors invading main branches of the portal vein. CONCLUSIONS: The present study has demonstrated the expression of NY-ESO-1 mRNA in hepatocellular carcinoma and NY-ESO-1 antibody production in patients with advanced hepatocellular carcinoma. Although the enhancement of NY-ESO-1 protein expression and the activation of immune response of the patients with hepatocellular carcinoma are necessary, NY-ESO-1 has the potential to be a good target molecule for immunotherapy against advanced hepatocellular carcinoma.


Subject(s)
Antigens, Neoplasm/genetics , Carcinoma, Hepatocellular/genetics , Liver Neoplasms/genetics , Membrane Proteins/genetics , Adult , Aged , Antigens, Neoplasm/immunology , Antigens, Neoplasm/metabolism , Carcinoma, Hepatocellular/immunology , Carcinoma, Hepatocellular/metabolism , DNA, Neoplasm/analysis , Female , Gene Expression , Humans , Immunohistochemistry , Liver/pathology , Liver Neoplasms/immunology , Liver Neoplasms/metabolism , Male , Membrane Proteins/immunology , Membrane Proteins/metabolism , Middle Aged , Reverse Transcriptase Polymerase Chain Reaction
15.
Cancer Sci ; 96(11): 801-9, 2005 Nov.
Article in English | MEDLINE | ID: mdl-16271074

ABSTRACT

For regulatory factor X4 (RFX4), two alternatively spliced variants, RFX4-A and -B, were reported in the testis. In this study, we identified transcript variants RFX4-C, -D, -E, and -F, and demonstrated by reverse transcription-polymerase chain reaction (RT-PCR) that RFX4-A, -B and -C mRNAs were expressed only in the testis, and RFX4-D mRNA was expressed only in normal brain tissues. In tumors, RFX4-E and -F in addition to RFX4-D mRNA were expressed in gliomas by rapid amplification of cDNA ends and RT-PCR analyses. Expression of RFX4 mRNA was not observed in other tumors, such as lung, esophageal, stomach, colon or liver cancers. Quantitative real-time RT-PCR using common primer pairs detecting all of the variant transcripts showed high expression in normal testis, low expression in the brain (1% compared to the expression in testis), and overexpression in 17 of 61 gliomas (28%). Western blot analysis using DC28 monoclonal antibody (mAb) produced against recombinant RFX4-D C-terminus protein showed expression of RFX4-A and -C proteins, but not RFX4-B protein, in the testis, and expression of RFX4-D protein in the brain. Moreover, expression of RFX4-E and -F proteins, but not RFX4-D protein, was observed in gliomas. Immunohistochemistry analysis using DC28 mAb showed positive staining in the nuclei of spermatocytes in the testis and glioma cells. Antibody against RFX4 was detected in the sera of 3 of 58 (5%) glioma patients by enzyme-linked immunosorbent assay, suggesting the immunogenicity of RFX4-E and -F proteins in glioma patients.


Subject(s)
Antibody Formation , Brain Neoplasms/immunology , Glioma/immunology , Transcription Factors/biosynthesis , Adolescent , Adult , Aged , Aged, 80 and over , Alternative Splicing , Blotting, Western , Brain/physiology , Child , Child, Preschool , Enzyme-Linked Immunosorbent Assay , Female , Gene Expression Profiling , Humans , Immunohistochemistry , Male , Middle Aged , Protein Isoforms , Regulatory Factor X Transcription Factors , Reverse Transcriptase Polymerase Chain Reaction , Testis/physiology
16.
Microbiol Immunol ; 49(11): 1009-16, 2005.
Article in English | MEDLINE | ID: mdl-16301813

ABSTRACT

OY-TES-1 was identified as a human homologue of the mouse, guinea pig, and pig proacrosin binding protein sp32 precursor. Differential expression levels of OY-TES-1 mRNA between testis and other normal tissues, and its expression in cancers indicated that OY-TES-1 would be classified as a cancer/testis antigen and considered to be a candidate of target antigen for cancer immunotherapy. In this study, we showed identification of HLA-A24-binding OY-TES-1 peptide, TES(401-409) (KTPFVSPLL) recognized by CD8 T-cells. Purified CD8 T-cells from healthy donors stimulated in vitro with the peptide-pulsed autologous DC and PBMC produced IFNgamma in response to the peptide-pulsed PBMC and showed cytotoxicity against the peptide-pulsed autologous EBV-B specifically. Furthermore, cytotoxicity was also observed against an OY-TES-1 mRNA-expressing tumor line, LK79. The retention time of the fraction in HPLC of the acid eluate from LK79 cells that showed positive sensitization against autologous EBV-B cells in recognition by CD8 CTL was the same as that of the fraction of the TES(401-409) peptide itself, suggesting that the TES(401-409) was a naturally processed peptide on LK79.


Subject(s)
CD8-Positive T-Lymphocytes/immunology , Carrier Proteins/immunology , HLA-A Antigens/metabolism , Oligopeptides/pharmacology , T-Lymphocytes, Cytotoxic/immunology , Animals , Antigens, Neoplasm/genetics , Antigens, Neoplasm/immunology , Cell Line, Tumor , Epitopes , HLA-A24 Antigen , Humans , Mice
17.
J Biosci Bioeng ; 99(1): 38-42, 2005 Jan.
Article in English | MEDLINE | ID: mdl-16233751

ABSTRACT

The PHT1 promoter::GUS fusion gene was constructed and introduced into Arabidopsis and rice by Agrobacterium-mediated transformation. Strong beta-glucuronidase (GUS) activity was detected in roots and showed phosphate starvation induction both in Arabidopsis and rice. In contrast, GUS activity in aerial tissues such as those of the leaf and stem was low. In situ GUS staining of root tissue indicated that PHT1 was expressed in root hairs and the outer layer of the main roots, but not in root tips. The PHT1 promoter has a desirable character for biotechnological transgene expression in monocot rice plants.


Subject(s)
Arabidopsis Proteins/genetics , Arabidopsis Proteins/metabolism , Oryza/genetics , Oryza/metabolism , Phosphate Transport Proteins/genetics , Phosphate Transport Proteins/metabolism , Plant Roots/genetics , Plant Roots/metabolism , Protein Engineering/methods , Gene Expression Regulation, Plant/physiology , Gene Transfer Techniques , Plants, Genetically Modified/metabolism , Promoter Regions, Genetic/genetics , Recombinant Proteins/metabolism
18.
Clin Cancer Res ; 11(15): 5496-503, 2005 Aug 01.
Article in English | MEDLINE | ID: mdl-16061866

ABSTRACT

PURPOSE: XAGE-1 was originally identified by the search for PAGE/GAGE-related genes using expressed sequence tag database and was shown to exhibit characteristics of cancer/testis-like antigens. Four transcript variants XAGE-1a, XAGE-1b, XAGE-1c, and XAGE-1d have been identified thus far. We recently identified XAGE-1b as a dominant antigen recognized by sera from lung adenocarcinoma patients. We here investigated the mRNA expression of four XAGE-1 variants and XAGE-1 protein expression in non-small cell lung cancer (NSCLC). Humoral immune response to XAGE-1b was also evaluated in patients. EXPERIMENTAL DESIGN: Forty-nine NSCLC specimens were analyzed for the expression of four XAGE-1 transcript variants by conventional 30-cycle and real-time reverse transcription-PCR and XAGE-1 protein expression by immunohistochemistry. Sera from 74 patients were analyzed for XAGE-1b antibody production by ELISA and Western blot. RESULTS: XAGE-1b and XAGE-1d mRNA were detected in 15 and 6 of 49 lung cancer specimens, respectively. No XAGE-1a or XAGE-1c mRNA expression was observed. XAGE-1b mRNA expression was observed in 14 of 31 (45%) adenocarcinoma and 1 of 18 (6%) lung cancer with other histologic types. Immunohistochemical analysis using a XAGE-1 monoclonal antibody showed that 14 of 15 XAGE-1b mRNA-positive and 3 of 34 XAGE-1b mRNA-negative specimens expressed XAGE-1 protein. Seropositivity was observed in 5 of 56 patients with adenocarcinoma, whereas none of 18 patients with other histologic types produced XAGE-1b antibody. CONCLUSION: XAGE-1b is highly and strongly expressed in lung adenocarcinoma and immunogenic in patients, suggesting that XAGE-1b is a promising antigen for immunotherapy against lung adenocarcinoma.


Subject(s)
Antigens, Neoplasm/biosynthesis , Carcinoma, Non-Small-Cell Lung/metabolism , Lung Neoplasms/metabolism , Adenocarcinoma/metabolism , Adenocarcinoma/pathology , Antibodies, Monoclonal/chemistry , Antigens, Neoplasm/chemistry , Blotting, Western , DNA Primers/chemistry , DNA, Complementary/metabolism , Databases as Topic , Enzyme-Linked Immunosorbent Assay , Expressed Sequence Tags , Genetic Vectors , Humans , Immunohistochemistry , Immunotherapy/methods , Plasmids/metabolism , Protein Structure, Tertiary , RNA, Messenger/metabolism , Recombinant Proteins/chemistry , Recombinant Proteins/metabolism , Reverse Transcriptase Polymerase Chain Reaction , Treatment Outcome
19.
BMC Cancer ; 5: 9, 2005 Jan 20.
Article in English | MEDLINE | ID: mdl-15661074

ABSTRACT

BACKGROUND: Insulin-like growth factor binding protein (IGFBP)-3 functions as a carrier of insulin-like growth factors (IGFs) in circulation and a mediator of the growth suppression signal in cells. There are two reported p53 regulatory regions in the IGFBP3 gene; one upstream of the promoter and one intronic. We previously reported a hot spot of promoter hypermethylation of IGFBP-3 in human hepatocellular carcinomas and derivative cell lines. As the hot spot locates at the putative upstream p53 consensus sequences, these p53 consensus sequences are really functional is a question to be answered. METHODS: In this study, we examined the p53 consensus sequences upstream of the IGFBP-3 promoter for the p53 induced expression of IGFBP-3. Deletion, mutagenesis, and methylation constructs of IGFBP-3 promoter were assessed in the human hepatoblastoma cell line HepG2 for promoter activity. RESULTS: Deletions and mutations of these sequences completely abolished the expression of IGFBP-3 in the presence of p53 overexpression. In vitro methylation of these p53 consensus sequences also suppressed IGFBP-3 expression. In contrast, the expression of IGFBP-3 was not affected in the absence of p53 overexpression. Further, we observed by electrophoresis mobility shift assay that p53 binding to the promoter region was diminished when methylated. CONCLUSION: From these observations, we conclude that four out of eleven p53 consensus sequences upstream of the IGFBP-3 promoter are essential for the p53 induced expression of IGFBP-3, and hypermethylation of these sequences selectively suppresses p53 induced IGFBP-3 expression in HepG2 cells.


Subject(s)
Gene Expression Regulation, Neoplastic , Gene Silencing , Insulin-Like Growth Factor Binding Protein 3/genetics , Promoter Regions, Genetic , Tumor Suppressor Protein p53/metabolism , Base Sequence , Binding Sites , Carcinoma, Hepatocellular/genetics , Cell Line, Tumor , Consensus Sequence , DNA Methylation , Electrophoretic Mobility Shift Assay , Humans , Liver Neoplasms/genetics , Molecular Sequence Data , Mutagenesis, Site-Directed , Sequence Deletion
20.
Int J Oncol ; 26(1): 57-63, 2005 Jan.
Article in English | MEDLINE | ID: mdl-15586225

ABSTRACT

We investigated NY-ESO-1 and LAGE-1a mRNA expression in normal tissues and various types of cancer by quantitative real-time RT-PCR. In addition to their high expression in the testis, we observed a low expression of NY-ESO-1 mRNA in the placenta, pancreas and liver, and no expression in 12 other normal tissues. We also observed a low expression of LAGE-1a mRNA in the placenta and ovary, and marginal expression in 13 other normal tissues. In contrast to the previous finding that NY-ESO-1 and LAGE-1a mRNAs were mostly co-expressed in solid tumors, we found an independent expression of NY-ESO-1 and LAGE-1a mRNAs. NY-ESO-1 mRNA expression was mostly associated with LAGE-1a mRNA expression in esophageal and liver cancers, but not in prostate cancer. Immunohistochemistry (IHC) using NY-ESO-1-specific ES121 mAb showed that NY-ESO-1 protein was detected in 6 of 9 and 3 of 10 NY-ESO-1 mRNA-positive specimens from esophageal and liver cancers, respectively. NY-ESO-1 protein expression was correlated with the copy numbers of NY-ESO-1 mRNA. IHC was also performed using ES121 mAb and B9.8 mAb recognizing both NY-ESO-1 and LAGE-1a in 4 esophageal and 6 liver cancer specimens preferentially expressing LAGE-1a mRNA. B9.8-specific staining was observed weakly and focally in one liver cancer specimen expressing >10(5) copies of LAGE-1a mRNA.


Subject(s)
Antigens, Neoplasm/metabolism , Membrane Proteins/metabolism , Neoplasms/metabolism , Antibodies, Monoclonal/immunology , Antigens, Neoplasm/analysis , Antigens, Neoplasm/genetics , Female , Humans , Liver/metabolism , Male , Membrane Proteins/analysis , Membrane Proteins/genetics , Neoplasms/genetics , Neoplasms/immunology , Ovary/metabolism , Placenta/metabolism , Protein Biosynthesis/genetics , RNA, Messenger/analysis , RNA, Messenger/metabolism , Reverse Transcriptase Polymerase Chain Reaction , Testis/metabolism , Tissue Distribution/genetics
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