Dual channel STED nanoscopy of lytic granules on actin filaments in natural killer cells.
AuthorsMace EM, Orange JS,
JournalCommun Integr Biol
PubMed ID22808328
Natural killer (NK) cells are innate immune effectors that eliminate diseased and tumorigenic targets through the directed secretion of specialized secretory lysosomes, termed lytic granules. This directed secretion is triggered following the formation of an immunological synapse (IS), which is characterized by actin re-modeling and receptor organization at the interface ... More
Tyramide signal amplification for analysis of kinase activity by intracellular flow cytometry.
'Intracellular flow cytometry permits quantitation of diverse molecular targets at the single-cell level. However, limitations in detection sensitivity inherently restrict the method, sometimes resulting in the inability to measure proteins of very low abundance or to differentiate cells expressing subtly different protein concentrations. To improve these measurements, an enzymatic amplification ... More
Dual receptor T cells mediate pathologic alloreactivity in patients with acute graft-versus-host disease.
AuthorsMorris GP, Uy GL, Donermeyer D, Dipersio JF, Allen PM,
Journal
PubMed ID23740900
Acute graft-versus-host disease (aGVHD) results from a robust response of donor T cells transferred during hematopoietic stem cell transplantation (HSCT) to allogeneic peptide-major histocompatibility complex antigens. Previous investigations have not identified T cell subsets that selectively mediate either protective immunity or pathogenic alloreactivity. We demonstrate that the small subset of ... More
Activation of T lymphocytes in atherosclerotic plaques.
AuthorsGrivel JC, Ivanova O, Pinegina N, Blank PS, Shpektor A, Margolis LB, Vasilieva E,
JournalArterioscler Thromb Vasc Biol
PubMed ID21960562
To decipher the immunologic mechanisms of plaque maturation and rupture, it is necessary to analyze the phenotypes and distribution of individual lymphocytes that migrate to the plaques, as well as their activation at different stages of plaque formation. We developed a protocol to isolate plaque-residing immune cells and analyze their ... More
In vivo functional analysis and genetic modification of in vitro-derived mouse neutrophils.
AuthorsMcDonald JU, Cortini A, Rosas M, Fossati-Jimack L, Ling GS, Lewis KJ, Dewitt S, Liddiard K, Brown GD, Jones SA, Hallett MB, Botto M, Taylor PR,
JournalFASEB J
PubMed ID21368104
Mature neutrophils are notoriously short-lived immune cells that cannot be genetically manipulated. Analysis of gene function therefore requires genetically modified animals, which is expensive, time-consuming, and costly in animal life. Analysis of gene function in neutrophils in a physiologically relevant context thus represents a significant problem in the field. We ... More
T cell-signaling network analysis reveals distinct differences between CD28 and CD2 costimulation responses in various subsets and in the MAPK pathway between resting and activated regulatory T cells.
AuthorsKalland ME, Oberprieler NG, Vang T, Taskén K, Torgersen KM,
JournalJ Immunol
PubMed ID22013130
To uncover signaling system differences between T cell stimuli and T cell subsets, phosphorylation status of 18 signaling proteins at six different time points following TCR triggering and CD28/CD2 costimulation was examined in human T cell subsets by phospho-epitope-specific flow cytometry of fluorescent cell barcoded samples, thereby providing a high-resolution ... More
B-cell signaling networks reveal a negative prognostic human lymphoma cell subset that emerges during tumor progression.
Human tumors contain populations of both cancerous and host immune cells whose malignant signaling interactions may define each patient's disease trajectory. We used multiplexed phospho-flow cytometry to profile single cells within human follicular lymphoma tumors and discovered a subpopulation of lymphoma cells with impaired B cell antigen receptor (BCR) signaling. ... More