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91.
Sullivan NL Eickhoff CS Zhang X Giddings OK Lane TE Hoft DF 《Journal of immunology (Baltimore, Md. : 1950)》2011,187(3):1358-1368
Trypanosoma cruzi is an intracellular parasite and the causative agent of Chagas disease. Previous work has shown that the chemokine receptor CCR5 plays a role in systemic T. cruzi protection. We evaluated the importance of CCR5 and CCL5 for mucosal protection against natural oral and conjunctival T. cruzi challenges. T. cruzi-immune CCR5(-/-) and wild-type C57BL/6 mice were generated by repeated infectious challenges with T. cruzi. CCR5(-/-) and wild-type mice developed equivalent levels of cellular, humoral, and protective mucosal responses. However, CCR5(-/-)-immune mice produced increased levels of CCL5 in protected gastric tissues, suggesting compensatory signaling through additional receptors. Neutralization of CCL5 in CCR5(-/-)-immune mice resulted in decreased mucosal inflammatory responses, reduced T. cruzi-specific Ab-secreting cells, and significantly less mucosal T. cruzi protection, confirming an important role for CCL5 in optimal immune control of T. cruzi replication at the point of initial mucosal invasion. To investigate further the mechanism responsible for mucosal protection mediated by CCL5-CCR5 signaling, we evaluated the effects of CCL5 on B cells. CCL5 enhanced proliferation and IgM secretion in highly purified B cells triggered by suboptimal doses of LPS. In addition, neutralization of endogenous CCL5 inhibited B cell proliferation and IgM secretion during stimulation of highly purified B cells, indicating that B cell production of CCL5 has important autocrine effects. These findings demonstrate direct effects of CCL5 on B cells, with significant implications for the development of mucosal adjuvants, and further suggest that CCL5 may be important as a general B cell coactivator. 相似文献
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New Tetrahymena basal body protein components identify basal body domain structure 总被引:4,自引:0,他引:4
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Kilburn CL Pearson CG Romijn EP Meehl JB Giddings TH Culver BP Yates JR Winey M 《The Journal of cell biology》2007,178(6):905-912
Basal bodies organize the nine doublet microtubules found in cilia. Cilia are required for a variety of cellular functions, including motility and sensing stimuli. Understanding this biochemically complex organelle requires an inventory of the molecular components and the contribution each makes to the overall structure. We define a basal body proteome and determine the specific localization of basal body components in the ciliated protozoan Tetrahymena thermophila. Using a biochemical, bioinformatic, and genetic approach, we identify 97 known and candidate basal body proteins. 24 novel T. thermophila basal body proteins were identified, 19 of which were localized to the ultrastructural level, as seen by immunoelectron microscopy. Importantly, we find proteins from several structural domains within the basal body, allowing us to reveal how each component contributes to the overall organization. Thus, we present a high resolution localization map of basal body structure highlighting important new components for future functional studies. 相似文献
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Mark Winey Janet B. Meehl Eileen T. O'Toole Thomas H. Giddings Jr. 《Molecular biology of the cell》2014,25(3):319-323
Researchers have used transmission electron microscopy (TEM) to make contributions to
cell biology for well over 50 years, and TEM continues to be an important technology in
our field. We briefly present for the neophyte the components of a TEM-based study,
beginning with sample preparation through imaging of the samples. We point out the
limitations of TEM and issues to be considered during experimental design. Advanced
electron microscopy techniques are listed as well. Finally, we point potential new users
of TEM to resources to help launch their project.Transmission electron microscopy (TEM) has been an important technology in cell biology ever
since it was first used in the early 1940s. The most frequently used TEM application in cell
biology entails imaging stained thin sections of plastic-embedded cells by passage of an
electron beam through the sample such that the beam will be absorbed and scattered, producing
contrast and an image (see Term Definition Beem capsule Plastic forms that hold samples in resin during polymerization Blocks (bullets) Polymerized samples in plastic removed from the Beem capsule and ready
to section Block face Small surface trimmed on a block before sectioning Boat Water reservoir in which sections float after being cut by a knife CLEM Correlative light and electron microscopy Dehydration Removal of water from a sample by replacement with solvent Electron tomography (ET) A method to image thick sections (200–300 nm) and produce
three-dimensional images Embedding Process of infiltrating the sample with resin Fixation Sample preservation with low temperature and/or chemicals to maintain
sample integrity Grid Small metal support that holds the sections for viewing in the electron
microscope HPF/FS High-pressure freezing/freeze substitution sample preparation
technique Immuno-EM Detection of proteins in EM samples using antibodies In-FXXKing credible!!!! Actual user quote in response to particularly beautiful sample. You may
embellish with your own words. Knife A very sharp edge, either glass or diamond, used to slice off
resin-embedded samples into sections Pre-embedding labeling Application of antibodies before fixation and embedding Post-embedding labeling Application of antibodies to sections on the grid Poststaining Staining with heavy metals of sections on a grid Resin Liquid form of the plastics used for embedding Ribbon Collection of serial sections placed on the grid Serials sections One-after-the-other thin sections in a ribbon TEM Transmission electron microscopy Thin sections The 60- to 70-nm sections cut from the samples in blocks Trimming Process of cutting away excess resin to create a block face Ultramicrotome Instrument used to cut sections Vitrification/vitreous ice Unordered ice in which samples can be viewed without fix or stain