CD8 T cell competition for dendritic cells in vivo is an early event in activation.
AuthorsWillis RA, Kappler JW, Marrack PC
JournalProc Natl Acad Sci U S A
PubMed ID16880405
'T cell responses against an antigen are often focused on a small fraction of potentially immunogenic determinants, a phenomenon known as immunodominance. Immunodominance can be established at several stages of antigen presentation, including antigen processing, binding of peptides to MHC, and competition between T cells for dendritic cells (DCs). The ... More
Color-blind fluorescence detection for four-color DNA sequencing.
AuthorsLewis EK, Haaland WC, Nguyen F, Heller DA, Allen MJ, MacGregor RR, Berger CS, Willingham B, Burns LA, Scott GB, Kittrell C, Johnson BR, Curl RF, Metzker ML
JournalProc Natl Acad Sci U S A
PubMed ID15800037
'We present an approach called pulsed multiline excitation (PME) for measurements of multicomponent, fluorescence species and demonstrate its application in capillary electrophoresis for DNA sequencing. To fully demonstrate the advantages of PME, a fluorescent dye set has been developed whose absorption maxima span virtually the entire visible spectrum. Unlike emission ... More
Multicolor super-resolution imaging with photo-switchable fluorescent probes.
AuthorsBates M, Huang B, Dempsey GT, Zhuang X,
JournalScience
PubMed ID17702910
Recent advances in far-field optical nanoscopy have enabled fluorescence imaging with a spatial resolution of 20 to 50 nanometers. Multicolor super-resolution imaging, however, remains a challenging task. Here, we introduce a family of photo-switchable fluorescent probes and demonstrate multicolor stochastic optical reconstruction microscopy (STORM). Each probe consists of a photo-switchable ... More
Correlative live-cell and superresolution microscopy reveals cargo transport dynamics at microtubule intersections.
AuthorsBálint Š, Verdeny Vilanova I, Sandoval Álvarez Á, Lakadamyali M,
JournalProc Natl Acad Sci U S A
PubMed ID23401534
Intracellular transport plays an essential role in maintaining the organization of polarized cells. Motor proteins tether and move cargos along microtubules during long-range transport to deliver them to their proper location of function. To reach their destination, cargo-bound motors must overcome barriers to their forward motion such as intersection points ... More
Cascade blue as a donor for resonance energy transfer studies of heme-containing proteins.
Cascade Blue acetyl azide is an amine reactive compound with spectral properties ideally suited for fluorescence resonance energy transfer (FRET) studies in which heme prosthetic groups serve as acceptors. To demonstrate utility of the Cascade Blue-heme spectroscopic ruler, cytochrome c was employed as a test case to calibrate distance measurements ... More
Rapid generation of fully human monoclonal antibodies specific to a vaccinating antigen.
AuthorsSmith K, Garman L, Wrammert J, Zheng NY, Capra JD, Ahmed R, Wilson PC,
JournalNat Protoc
PubMed ID19247287
We describe herein a protocol for the production of antigen-specific human monoclonal antibodies (hmAbs). Antibody-secreting cells (ASCs) are isolated from whole blood collected 7 d after vaccination and sorted by flow cytometry into single cell plates. The antibody genes of the ASCs are then amplified by RT-PCR and nested PCR, ... More
Rab conversion as a mechanism of progression from early to late endosomes.
AuthorsRink J, Ghigo E, Kalaidzidis Y, Zerial M
JournalCell
PubMed ID16143105
The mechanisms of endosome biogenesis and maintenance are largely unknown. The small GTPases Rab 5 and Rab 7 are key determinants of early and late endosomes, organizing effector proteins into specific membrane subdomains. Whether such Rab machineries are indefinitely maintained on membranes or can disassemble in the course of cargo ... More