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61.
Rapid and efficient generation of lentivirally gene-modified dendritic cells from DC progenitors with bone marrow stromal cells 总被引:4,自引:0,他引:4
Sumimoto H Tsuji T Miyoshi H Hagihara M Takada-Yamazaki R Okamoto S Ikeda Y Takahashi T Kawakami Y 《Journal of immunological methods》2002,271(1-2):153-165
Since dendritic cells (DC) play pivotal roles in both innate and adaptive immunity, DC can be a good target for immuno-gene therapy. However, the optimal generation method for gene-modified DC has not yet been well exploited. CD34+ cells from cord blood (CB), bone marrow (BM), or peripheral blood (PB) were expanded in a medium containing stem cell factor (SCF), flt 3 ligand (Flt3L) and thrombopoietin (TPO) with or without HESS-5, a murine BM stromal cell line, for 2 weeks (the first expansion step), then differentiated to DC in a medium containing granulocyte-macrophage colony-stimulating factor (GM-CSF), flt 3 ligand (Flt3L), stem cell factor (SCF), tumor necrosis factor-alpha (TNF-alpha), IL-4, and lipopolysaccharide (LPS) for 9 days (the second differentiation step). DC progenitors were transduced with human immunodeficiency virus (HIV) vectors at different time points during the second step. Use of HESS-5 during the first step resulted in more DC generation than without it (cell expansion: CB, 10,461 vs. 354-fold; BM, 962 vs. 225-fold; peripheral blood mononuclear cell (PBMC), 8,506 vs. 240-fold; %DC: CB, 83.4% vs. 76.9%; BM, 83.6 vs. 69.8%; PBMC, 85.9 vs. 60.5%). Gene transduction to the in vitro expanded DC progenitors at day 3 during the second step, resulted in better final yield of the gene-modified DC than that to those at day 0 or day 6 (as much as 44% of DC expressed green fluorescence protein (GFP) as a transgene) and the transduction efficiency correlated with endocytic ability and percent of S phase. DC transduced with an HIV vector encoding a melanoma antigen, MART-1, were adequately recognized by specific anti-MART-1 CTL. The two-step culture method with HESS-5 is useful for rapid expansion of DC progenitors and subsequent lentiviral gene transduction to DC. 相似文献
62.
Okada K Sugiura T Kuroda E Tsuji S Yamashita U 《Clinical and experimental immunology》2001,124(3):406-413
The effects of chronic administration of phenytoin, a common anticonvulsive drug, on immune responses were studied in mice. Anti-keyhole limpet haemocyanin (KLH) IgE antibody response after KLH-immunization was enhanced in phenytoin-treated mice. Proliferative responses of spleen cells induced with KLH, concanavalin A (ConA), lipopolysaccharide and anti-CD3 antibody were reduced in phenytoin-treated mice. Accessory function of spleen adherent cells on ConA-induced T cell proliferative response was reduced in phenytoin-treated mice. KLH-induced IL-4 production of spleen cells was enhanced, while IFN-gamma production was reduced in phenytoin-treated mice. In addition, production of IL-1 alpha, but not IL-6 and IL-12 by spleen adherent cells from phenytoin-treated mice was reduced. Natural killer cell activity was reduced in phenytoin-treated mice. These results suggest that phenytoin treatment preferentially induces a Th2 type response. We also observed that plasma ACTH and corticosterone levels were increased in phenytoin-treated mice, and speculated that phenytoin might act directly and indirectly, through HPA axis activation, on the immune system to modulate Th1/Th2 balance. 相似文献
63.
Identification of Vibrio parahaemolyticus pandemic group-specific DNA sequence by genomic subtraction 总被引:1,自引:0,他引:1 下载免费PDF全文
Okura M Osawa R Arakawa E Terajima J Watanabe H 《Journal of clinical microbiology》2005,43(7):3533-3536
A genomic subtraction between a pandemic Vibrio parahaemolyticus and a nonpandemic strain that seemed to be clonally related was performed. A subtractive DNA fragment was identified to be a part of a 16-kbp insertion sequence which was present in almost all pandemic strains but not in nonpandemic strains tested. 相似文献
64.
In vivo expression of monokine and inducible nitric oxide synthase in experimentally induced pulmonary granulomatous inflammation. Evidence for sequential production of interleukin-1, inducible nitric oxide synthase, and tumor necrosis factor. 下载免费PDF全文
The present study examined the temporal pattern and localization of interleukin-1, tumor necrosis factor-alpha, and inducible nitric oxide synthase expression in lung tissue undergoing foreign body granuloma formation. Pulmonary granulomas were induced by the intratracheal injection of dextran beads into genetically high granuloma responder, carrying Bcgs (BALB/c), and low responder, carrying Bcgr (C3H/HeJ and DBA/2), mice. There was a pattern of sequential expression of these molecules in BALB/c mice. Thus, interleukin-1 alpha and inducible nitric oxide synthase were induced mostly in the cells accumulated around the beads and also in some bronchiolar epithelial cells during the early phase (1 to 3 days), whereas tumor necrosis factor-alpha was induced in the cells around the beads at the later resolution phase (3 to 7 days). By contrast, in low responder mice, an increase in the expression of interleukin-1 alpha and inducible nitric oxide synthase was detected in lung macrophages as well as in alveolar cells and bronchiolar epithelial cells on day 1, but that of tumor necrosis factor-alpha was not detected throughout the period. These results together with our previous findings on cytokine activity in granuloma extract suggest that interleukin-1 and nitric oxide produced by recruited macrophages may take part in the early, macrophage-dependent phase of granuloma formation whereas tumor necrosis factor-alpha may be more crucial as a mediator responsible for the difference in innate resistance or susceptibility to granuloma formation. 相似文献
65.
Depression of the generation of cell-mediated cytotoxicity by suppressor cells after surgery 下载免费PDF全文
S Miyazaki T Akiyoshi S Arinaga F Koba T Wada H Tsuji 《Clinical and experimental immunology》1983,54(2):573-579
The effects of surgical operation on the generation of cell-mediated cytotoxicity in mixed cell cultures were studied in patients with various carcinomas or benign lesions. Peripheral blood mononuclear cells from patients were cultured with B lymphoblastoid cell line Raji in mixed culture, and the induced cytotoxicity was measured by 51Cr release assay. In 15 patients with various carcinomas, the capacity of cells to generate cytotoxic cells was significantly depressed 1, 3 and 6 days after surgery, as compared to that before surgery. It returned to the pre-operative level by the 8th post-operative day. In eight patients with benign lesions, significant decrease in cytotoxic cell activity was observed 3 and 6 days after operation. At the 8th day, however, there was a significant increase in the generated cytotoxicity. The depressed generation of cytotoxic cells 3 days after surgery could be abrogated by removal of adherent cells from the responding cell population. This effect could be partially reconstituted by addition of separated, autologous adherent cells back to the responding non-adherent cell culture. These results demonstrate that suppressor cells, presumably monocytes, may be responsible for the depressed generation of cytotoxic cells after surgery. 相似文献
66.
Enkhtuvshin Gereltzul Yoshiyuki Baba Naoto Suda Momotoshi Shiga Maristela Sayuri Inoue Michiko Tsuji Insik Shin Yukio Hirata Kimie Ohyama Keiji Moriyama 《Journal of human genetics》2008,53(10):941-946
This is a report of a 27-year-old woman with an unusual de novo chromosomal abnormality. Mosaicism was identified in peripheral
blood cells examined by standard G-bands by trypsin using Giemsa (GTG) analysis and fluorescence in situ hybridization (FISH)
analysis with chromosome-18 region-specific probes, 46,XX,del(18)(pter → q21.33:)[41], 46,XX,r(18)(::p11.21 → q21.33::)[8],
and 46,XX,der(18)(pter → q21.33::p11.21 → pter)[1]. On the other hand, the karyotype of periodontal ligament fibroblasts was
nonmosaic, 46,XX, der(18)(pter → q21.33::p11.21 → pter)[50]. All cell lines appeared to be missing a portion of 18q (q21.33 → qter).
The pattern of the dup(18p)/del(18q) in the rod configuration raises the possibility of an inversion in chromosome 18 in one
of the parents. However, no chromosomal anomaly was detected in either parent. The most probable explanation is that de novo
rod and ring configurations arose simultaneously from an intrachromosomal exchange. The unique phenotype of this patient,
which included primary hypothyroidism and primary hypogonadism, is discussed in relation to her karyotype. 相似文献
67.
Renal functions were examined in 102 patients with yusho in 1988, Frequencies of proteinuria, microhematuria and history of renal diseases were not different from 20 age-matched controls. The means of blood urea nitrogen, serum creatinine and serum uric acid levels of yusho patients did not differ from those of controls. The levels of serum beta 2-microglobulin and its urinary excretion showed no difference between two groups. Serum concentrations of sodium, potassium, chloride, calcium and phosphorus revealed no abnormality in all patients except for one who had hypophosphatemia. Urinary excretions of phosphorus, however, were significantly higher in yusho patients than in controls. Serum PCB levels, which were still higher in yusho patients, did not correlate with urinary excretions of phosphorus. The mechanism and the clinical significance of this phenomenon remain to be elucidated. 相似文献
68.
Modified PCR-RFLP method for HLA-DPB1 and -DQA1 genotyping. 总被引:17,自引:0,他引:17
M Ota T Seki N Nomura K Sugimura N Mizuki H Fukushima K Tsuji H Inoko 《Tissue antigens》1991,38(2):60-71
We previously developed a new technique for HLA class II genotyping by digestion of polymerase chain reaction-amplified genes with restriction endonucleases (PCR-RFLP method). This PCR-RFLP method is an efficient and convenient typing technique for class II alleles. However, small fragments or bands located close to each other on polyacrylamide gels sometimes prevent precise analysis of the RFLP bands. Furthermore, the restriction enzymes we have reported in the previous papers are not sufficient to identify the genotypes of all heterozygous individuals. Here, we report an improved PCR-RFLP method using some informative restriction enzymes which have either a single cleavage site or, alternatively, no cleavage site in the amplified DNA region, depending on the HLA alleles, making reading of RFLP band patterns much easier. Each second exon of the HLA-DQA1 or -DPB1 gene was selectively amplified from genomic DNAs of 70 HLA-homozygous B-cell lines and 100 healthy Japanese by PCR. Amplified DNAs were digested with restriction endonucleases and then subjected to electrophoresis assaying simply for cutting, or no cutting, of the DNA. ApaLI, HphI, BsaJI, FokI, MboII and Mn1I can discriminate eight alleles of the DQA1 gene. Similarly 19 alleles of the DPB1 gene can be discriminated with Bsp1286I, FokI, DdeI, BsaJI, BssHII, Cfr13I, RsaI, EcoNI, and AvaII enzymes. This modified PCR-RFLP method can be successfully applied to heterozygotes. Thus, the method is technically simpler and more practical for routine HLA typing work than our previous PCR-RFLP method. 相似文献
69.
70.
Isolation of amantadine-resistant influenza a viruses (H3N2) from patients following administration of amantadine in Japan 总被引:3,自引:0,他引:3 下载免费PDF全文
Iwahashi J Tsuji K Ishibashi T Kajiwara J Imamura Y Mori R Hara K Kashiwagi T Ohtsu Y Hamada N Maeda H Toyoda M Toyoda T 《Journal of clinical microbiology》2001,39(4):1652-1653
In Japan, the use of amantadine for treatment of influenza A virus infection was not accepted until November 1998, although it was widely used for treatment of Parkinsonism. Since then, we have monitored the emergence of amantadine-resistant viruses and isolated two viruses from patients on long-term treatment with amantadine. 相似文献