Detail of "34215-57-1"

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Discriminating in vitro cell fusion from cell aggregation by flow cytometry combined with fluorescence resonance energy transfer

All Rights Reserved. Discriminating in vitro cell fusion from cell aggregation by flow cytometry combined with fluorescence resonance energy transfer. Huerta, Leonor; Lopez-Balderas, Nayali; Larralde, Carlos; Lamoyi, Edmundo (Ciudad Universitaria, Department of Immunology, Instituto de Investigaciones Biomedicas, Universidad Nacional Autonoma de Mexico, Distrito Federal, C.P. C.P. 04510, Mex.). Journal of Virological Methods, 138(1-2), 17-23 (English) 2006 Elsevier B.V. CODEN: JVMEDH. ISSN: 0166-0934. DOCUMENT TYPE: Journal CA Section: 9 (Biochemical Methods) Expression of fusion proteins in the plasma membrane enables cells to bind and fuse with surrounding cells to form syncytia. Cell fusion can have important functional outcomes for the interacting cells, as syncytia formation does in AIDS pathogenesis. Studies on cell fusion would be facilitated by a quant. method able to discriminate between cellular aggregates and bona fide fused cells in a cell population.There are some commonly used reagents with their cas registry numbers 41085-99-8 and 34215-57-1 in this article. Flow cytometry with fluorescence resonance energy transfer is applied here for analyzing fusion of HIV-1 envelope-expressing cells with CD4+ Jurkat cells. Fusion partners were labeled with the vital lipophilic fluorescent probes DiO (green) and DiI (red) and FRET is manifested by an enhancement of the DiI red fluorescence intensity in double fluorescent cells, thus allowing discrimination between fused and aggregated cells. The inhibitory effect of anti-CD4 monoclonal antibodies and the inhibitory peptide T-20 upon cell fusion were readily quantified by this technique. This method allows the distinction of fused and aggregated cells even when they are at low frequencies. .

Preliminary study on natural killer T cells proliferation in vitro from human peripheral blood cells and their functions

All Rights Reserved. Preliminary study on natural killer T cells proliferation in vitro from human peripheral blood cells and their functions. Fan, Hua-hua; Zheng, Bin; Liu, Yan; Gao, Li; Nie, Xiao-xuan; Gao, Feng; Lu, Hua-zhong (Shanghai Blood Center, Shanghai 200051, Peop. Rep. China). Xiandai Mianyixue, 25(5), 398-401 (Chinese) 2005 Shanghai Shi Mianyixue Yanjiusuo. CODEN: XMIIAT. DOCUMENT TYPE: Journal CA Section: 15 (Immunochemistry) This paper intended to establish a method to proliferate the natural killer T (NKT) cells in vitro and to investigate their functions. TCR Va24+/Vb11+ NKT cells were proliferated from human peripheral blood mononuclear cells (MNC) and purified T lymphocytes by several kinds of methods. The levels of IL-4, IFN-g and TNF-a secretions from NKT cells were detd. by flow cytometry, and the CD4+ and CD8+ cells were removed by using CD4/CD8 Dynal beads to further purify NKT cells. Meanwhile, the cytotoxic activity of NKT cells was measured by means of DIOC18 staining and flow cytometry. It was found that the NKT cells could be proliferated in vitro through the actions of a-Galcer and IL-2, and the ratio of TCR Va24+/Vb11+ NKT cells elevated up to 25.5% ± 7.2% after 19 days. The highest proliferation times of NKT cells might reach up to (1. 34215-57-1 and 17090-79-8 are cas registry numbers of chemicals which are used as reagents here.51 ± 0.91) X 104 times. The expanded NKT cells highly expressed TCR Va24, TCR Vb11, CD3 and CD161, while low expressed CD56, CD14 and CD19. Under the stimulation of CD3 monoclonal antibody and IL-2, the proportion of TCR Vb11+ cells secreting IL-4 and IFN-g was higher than that of the TCR Vb11- cells (P < 0.05). In addn., when the CD4+ and CD8+ cells were removed, the frequency of NKT cells increased to 80%. Intense cytotoxic effect of the NKT cells to tumor cell lines U937 and HL60 as well as to dendritic cells could be demonstrated. In conclusion, it is evident that the NKT cells can be expanded from human peripheral blood mononuclear cells directly, thus making simplification of the exptl. procedures. .