Features of medullary thymic epithelium implicate postnatal development in maintaining epithelial heterogeneity and tissue-restricted antigen expression.
AuthorsGillard GO, Farr AG
JournalJ Immunol
PubMed ID16670287
'Although putative thymic epithelial progenitor cells have been identified, the developmental potential of these cells, the extent of medullary thymic epithelium (mTEC) heterogeneity, and the mechanisms that mediate the expression of a wide range of peripheral tissue-restricted Ags (TRAs) by mTECs remain poorly defined. Here we have defined several basic ... More
Overexpression of the Wiskott-Aldrich syndrome protein N-terminal domain in transgenic mice inhibits T cell proliferative responses via TCR signaling without affecting cytoskeletal rearrangements.
AuthorsSato M, Tsuji NM, Gotoh H, Yamashita K, Hashimoto K, Tadotsu N, Yamanaka H, Sekikawa K, Hashimoto Y
JournalJ Immunol
PubMed ID11591801
'Wiskott-Aldrich syndrome (WAS) is an X-linked recessive disorder characterized by thrombocytopenia with small platelets, severe eczema, and recurrent infections due to defects in the immune system. The disease arises from mutations in the gene encoding the WAS protein (WASP), which plays a role as an adaptor molecule in signal transduction ... More
Reversible MHC multimer staining for functional isolation of T-cell populations and effective adoptive transfer.
AuthorsKnabel M, Franz TJ, Schiemann M, Wulf A, Villmow B, Schmidt B, Bernhard H, Wagner H, Busch DH
JournalNat Med
PubMed ID12042816
'Recently developed major histocompatibility complex (MHC) multimer technologies allow visualization and isolation of antigen-specific T cells. However, functional analysis and in vivo transfer of MHC multimer-stained cells is hampered by the persistence of T-cell receptor (TCR) MHC interactions and subsequently induced signaling events. As MHC monomers do not stably bind ... More
Detection of allergen-specific IgE on microarrays by use of signal amplification techniques.
AuthorsBacarese-Hamilton T, Mezzasoma L, Ingham C, Ardizzoni A, Rossi R, Bistoni F, Crisanti A
JournalClin Chem
PubMed ID12142398
Combined tyramide signal amplification and quantum dots for sensitive and photostable immunofluorescence detection.
AuthorsNess JM, Akhtar RS, Latham CB, Roth KA
JournalJ Histochem Cytochem
PubMed ID12871979
Conventional immunofluorescence detection of biologically relevant proteins and antigens in tissue sections is often limited by relatively weak signals that fade rapidly on illumination. We have developed an immunohistochemical protocol that combines the sensitivity of tyramide signal amplification with the photostability of quantum dots to overcome these limitations. This simple ... More
Turning blood into brain: cells bearing neuronal antigens generated in vivo from bone marrow.
AuthorsMezey E, Chandross KJ, Harta G, Maki RA, McKercher SR
JournalScience
PubMed ID11099419
Bone marrow stem cells give rise to a variety of hematopoietic lineages and repopulate the blood throughout adult life. We show that, in a strain of mice incapable of developing cells of the myeloid and lymphoid lineages, transplanted adult bone marrow cells migrated into the brain and differentiated into cells ... More
Visualization and identification of IL-7 producing cells in reporter mice.
AuthorsMazzucchelli RI, Warming S, Lawrence SM, Ishii M, Abshari M, Washington AV, Feigenbaum L, Warner AC, Sims DJ, Li WQ, Hixon JA, Gray DH, Rich BE, Morrow M, Anver MR, Cherry J, Naf D, Sternberg LR, McVicar DW, Farr AG, Germain RN, Rogers K, Jenkins NA, Copeland NG, Durum SK,
JournalPLoS One
PubMed ID19907640
Interleukin-7 (IL-7) is required for lymphocyte development and homeostasis although the actual sites of IL-7 production have never been clearly identified. We produced a bacterial artificial chromosome (BAC) transgenic mouse expressing ECFP in the Il7 locus. The construct lacked a signal peptide and ECFP (enhanced cyan fluorescent protein) accumulated inside ... More
A novel mechanism for protein delivery: granzyme B undergoes electrostatic exchange from serglycin to target cells.
AuthorsRaja SM, Metkar SS, Höning S, Wang B, Russin WA, Pipalia NH, Menaa C, Belting M, Cao X, Dressel R, Froelich CJ
JournalJ Biol Chem
PubMed ID15788411
The molecular interaction of secreted granzyme B-serglycin complexes with target cells remains undefined. Targets exposed to double-labeled granzyme B-serglycin complexes show solely the uptake of granzyme B. An in vitro model demonstrates the exchange of the granzyme from serglycin to immobilized, sulfated glycosaminoglycans. Using a combination of cell binding and ... More
OmpA targets dendritic cells, induces their maturation and delivers antigen into the MHC class I presentation pathway.
AuthorsJeannin P, Renno T, Goetsch L, Miconnet I, Aubry JP, Delneste Y, Herbault N, Baussant T, Magistrelli G, Soulas C, Romero P, Cerottini JC, Bonnefoy JY
JournalNat Immunol
PubMed ID11101872
We analyzed the interaction between a bacterial cell wall protein and dendritic cells (DCs). Outer membrane protein A from Klebsiella pneumoniae (kpOmpA) specifically bound to professional antigen presenting cells and was endocytosed by immature DCs via a receptor-dependent mechanism. kpOmpA signaled through Toll-like receptor 2, induced DCs to produce interleukin ... More
A microengraving method for rapid selection of single cells producing antigen-specific antibodies.
AuthorsLove JC, Ronan JL, Grotenbreg GM, van der Veen AG, Ploegh HL
JournalNat Biotechnol
PubMed ID16699501
Monoclonal antibodies that recognize specific antigens of interest are used as therapeutic agents and as tools for biomedical research. Discovering a single monoclonal antibody requires retrieval of an individual hybridoma from polyclonal mixtures of cells producing antibodies with a variety of specificities. The time required to isolate hybridomas by a ... More
Drosophila Squid/hnRNP helps Dynein switch from a gurken mRNA transport motor to an ultrastructural static anchor in sponge bodies.
AuthorsDelanoue R, Herpers B, Soetaert J, Davis I, Rabouille C,
JournalDev Cell
PubMed ID17925228
In Drosophila oocytes, dorso-anterior transport of gurken mRNA requires both the Dynein motor and the heterogeneous nuclear ribonucleoprotein (hnRNP) Squid. We show that gurken transcripts are transported directly on microtubules by Dynein in nonmembranous electron-dense transport particles that also contain Squid and the transport cofactors Egalitarian and Bicaudal-D. At its ... More
Inducible gene targeting in the neonatal vasculature and analysis of retinal angiogenesis in mice.
AuthorsPitulescu ME, Schmidt I, Benedito R, Adams RH,
JournalNat Protoc
PubMed ID20725067
The retina is a powerful experimental system for the analysis of angiogenic blood vessel growth in the postnatal organisms. The three-dimensional architecture of the vessel network and processes as diverse as endothelial cell (EC) proliferation, sprouting, perivascular cell recruitment, vessel remodeling or maturation can be investigated at high resolution. The ... More
Analysis of several fluorescent detector molecules for protein microarray use.
AuthorsWiese R
JournalLuminescence
PubMed ID12536376
The utility of several streptavidin-linked fluorescent detector molecules was evaluated on two protein microarray platforms. Tested detector molecules included: Alexa Fluor 546; R-phycoerythrin (RPE), orange fluospheres; Cy3-containing liposomes (Large Unilamellar Vesicles, LUV) labelled with Cy3; and an RPE-antibody complex. The two array architectures tested consisted of an array of murine ... More
Olfactory responses in a gustatory organ of the malaria vector mosquito Anopheles gambiae.
AuthorsKwon HW, Lu T, Rützler M, Zwiebel LJ
JournalProc Natl Acad Sci U S A
PubMed ID16938890
The proboscis is an important head appendage in insects that has primarily been thought to process gustatory information during food intake. Indeed, in Drosophila and other insects in which they have been identified, most gustatory receptors are expressed in proboscis neurons. Our previous characterization of the expression of AgOR7, a ... More
G-protein-coupled receptor activation induces the membrane translocation and activation of phosphatidylinositol-4-phosphate 5-kinase I alpha by a Rac- and Rho-dependent pathway.
AuthorsChatah NE, Abrams CS
JournalJ Biol Chem
PubMed ID11431481
Phosphatidylinositol 4,5-bisphosphate (PI4,5P(2)) mediates cell motility and changes in cell shape in response to extracellular stimuli. In platelets, it is synthesized from PI4P by PIP5K in response to stimulation of a G-protein-coupled receptor by an agonist, such as the thrombin. In the present study, we have addressed the pathway that ... More