81 research outputs found
Enhancement of the antigen-specific cytotoxic T lymphocyte-inducing ability in the PMDC11 leukemic plasmacytoid dendritic cell line via lentiviral vector-mediated transduction of the caTLR4 gene.
The aim of the present study was to enhance the efficiency of leukemia immunotherapy by increasing the antigen-specific cytotoxic T lymphocyte-inducing ability of leukemia cells. The leukemic plasmacytoid dendritic cell line PMDC05 containing the HLA-A02/24 antigen, which was previously established in our laboratory (Laboratory of Hematology and Oncology, Graduate School of Health Sciences, Niigata University, Niigata, Japan), was used in the present study. It exhibited higher expression levels of CD80 following transduction with lentiviruses encoding the CD80 gene. This CD80-expressing PMDC05 was named PMDC11. In order to establish a more potent antigen-presenting cell for cellular immunotherapy of tumors or severe infections, PMDC11 cells were transduced with a constitutively active (ca) toll-like receptor 4 (TLR4) gene using the Tet-On system (caTLR4-PMDC11). CD8(+) T cells from healthy donors with HLA-A02 were co-cultured with mutant WT1 peptide-pulsed PMDC11, lipopolysaccharide (LPS)-stimulated PMDC11 or caTLR4-PMDC11 cells. Interleukin (IL)-2 (50 IU/ml) and IL-7 (10 ng/ml) were added on day three of culture. Priming with mutant WT1 peptide-pulsed PMDC11, LPS-stimulated PMDC11 or caTLR4-PMDC11 cells was conducted once per week and two thirds of the IL-2/IL-7 containing medium was replenished every 3-4 days. Immediately prior to the priming with these various PMDC11 cells, the cultured cells were analyzed for the secretion of interferon (IFN)-γ in addition to the percentage and number of CD8(+)/WT1 tetramer(+) T cells using flow cytometry. caTLR4-PMDC11 cells were observed to possess greater antigen-presenting abilities compared with those of PMDC11 or LPS-stimulated PMDC11 cells in a mixed leukocyte culture. CD8 T cells positive for the WT1 tetramer were generated following 3-4 weeks of culture and CD8(+)/WT1 tetramer+ T cells were markedly increased in caTLR4-PMDC11-primed CD8(+) T cell culture compared with PMDC11 or LPS-stimulated PMDC11-primed CD8(+) T cell culture. These CD8(+) T cells co-cultured with caTLR4-PMDC11 cells were demonstrated to secrete IFN-γ and to be cytotoxic to WT1-expressing target cells. These data suggested that the antigen-specific cytotoxic T lymphocyte (CTL)-inducing ability of PMDC11 was potentiated via transduction of the caTLR4 gene. The present study also suggested that caTLR4-PMDC11 cells may be applied as potent antigen-presenting cells for generating antigen-specific CTLs in adoptive cellular immunotherapy against tumors and severe viral infections
Increased phosphatidylcholine (16:0/16:0) in the folliculus lymphaticus of Warthin tumor
Warthin tumor (War-T), the second most common benign salivary gland tumor, consists mainly of neoplastic epithelium and lymphoid stroma. Some proteins and genes thought to be involved in War-T were evaluated by molecular biology and immunology. However, lipids as an important component of many tumor cells have not been well studied in War-T. To elucidate the molecular biology and pathogenesis of War-T, we investigated the visualized distribution of phosphatidylcholines (PCs) by imaging mass spectrometry (IMS). In our IMS analysis of a typical case, 10 signals were significantly different in intensity (p < 0.01) between the War-T and non-tumor (Non-T) regions. Five specific PCs were frequently found in the War-T regions of all of the samples: [PC (16:0/16:0) + K](+) (m/z 772.5), [PC (16:0/20:4) + K](+) (m/z 820.5), [PC (16:0/20:3) + K](+) (m/z 822.5), [PC (18:2/20:4) + K](+) (m/z 844.5), and [PC (18:0/20:5) + K](+) (m/z 846.5). PC (16:0/16:0) was increased specifically in the folliculus lymphaticus of War-T lymphoid stroma, suggesting a different metabolism. Localization of PC (16:0/16:0) might reflect inflammation activity participating in the pathogenesis of War-T. Thus, our IMS analysis revealed the profile of PCs specific to the War-T region. The molecules identified in our study provide important information for further studies of War-T pathogenesis
Planning of observation and motion for interpretation of road intersection scenes considering uncertainty
A novel efficient feeder-free culture system for the derivation of human induced pluripotent stem cells
Nakagawa, M.et al. A novel efficient feeder-free culture system forthe derivation of human induced pluripotent stem cells.Sci. Rep.4, 3594; DOI:10.1038/srep03594 (2014)
胎盤における一酸化窒素(NO)の産生とNO合成酵素(NOS)の発現
Nitric oxide (NO) production in the rat placenta(pars fetalis placentae and decidua basalis) was monitored and quantified by electron paramagnetic resonance (EPR) spectroscopy using an iron complex with N-(dithiocarboxy) sarcosine (Fe-DTCS) as a NO trapping reagent. The expression of nitric oxide synthase (NOS) isoforms was also examined by quantitative reverse transcriptase-mediated polymerase chain reaction (RT-PCR) analysis. The EPR spectrum of the placenta with Fe-DTCS trapping showed a triplet signal (g = 2.038) derived from an NO-Fe-DTCS complex. The heights of the NO-Fe-DTCS (a) and MnO (b) signals were simultaneously measured to calculate the ratio (a/b) of these signal heights used for the quantification of the NO production level. Although, the ratio of the signal heights of NO-Fe-DTCS and MnO did not vary significantly with gestational stage in the fetal placenta, the ratio was markedly decreased in the maternal placenta during the last few days of gestation. At the gestational stages examined, the level of NOS 2 (iNOS) mRNA expression was significantly higher than that of NOS 3 (eNOS) mRNA expression at any given stage in the fetal placenta, and the production pattern of NO was in good accordance with the expression pattern of NOS 2 in the maternal placenta. These results suggest that NOS 2 is the predominant producer of NO in the placenta and that NOS 2-generated NO plays significant roles in the maintenance of placental functions immediately before birth
p63 regulates Satb1 to control tissue-specific chromatin remodeling during development of the epidermis
Genome organizer Satb1 is regulated by p63 and contributes to epidermal morphogenesis by remodeling chromatin structure and gene expression at the epidermal differentiation complex locus
ドッキョウ イカ ダイガク コシガヤ ビョウイン ニオケル, フクブ チョウ オンパ ケンサ ニヨル タンノウ リュウキセイ ビョウヘン ノ ケントウ
腹部超音波検査が施行された3572 例を対象として胆嚢隆起性病変の検討を行った.胆嚢隆起性病変は3572例中791例( 22.1%) に認められ,重複検査例を除いた773例の平均年齢は59.6±13.6歳であり,男性370 例,女性403 例であった.胆嚢隆起性病変の最大径の平均は4.7±5.8 mm で,単発が256 例 (33.1%),多発が517例( 66.9%) であった.773例中,10 mm 以上の病変を有する症例は44 例( 5.6%) であった.これら44例の最終診断は,胆嚢良性ポリープ19例( 43.2%),胆嚢腺筋症2 例( 4.6%),胆泥貯留2 例( 4.6%),胆嚢結石2例( 4.6%) 切除可能胆嚢癌6例( 13.6%),切除不能胆嚢癌6 例( 13.6%),その他の癌2 例( 4.6%),不明5例( 11.3%) であり,胆嚢癌の半数が切除不能であった.今後,超音波検査を用いて切除可能な胆嚢癌をより多く拾い上げるためには,人間ドック等による,より幅広いスクリーニングが必要であると考えられた.The present study investigated the presence and characteristicsof elevated gallbladder lesions in 3572 patients whounderwent abdominal ultrasonography in our hospital betweenApril 2011 and March 2012. Elevated gallbladder lesionswere present in 791 patients (22.1 %). After excludingpatients who underwent repeat examination, 44 of theremaining 773 patients (5.6 %) had lesions ≥ 10 mm. Finaldiagnoses in these 44 patients were as follows:benign gallbladderpolyp, n=19 (43.2 %);gallbladder adenomyosis,n=2 (4.6 %);biliary sludge accumulation, n=2 (4.6 %);gallbladder stone, n=2( 4.6%);resectable gallbladder cancer,n=6( 13.6%);non-resectable gallbladder cancer, n=6(13.6%);other cancers, n=2( 4.6%);and unknown, n=5(11.3 %). Wider screening during routine medical examinationssuch as annual health checks is required to enable increasedidentification of gallbladder cancer at an early stagewhen resection is still possible
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