Hematopoietic potential of stem cells isolated from murine skeletal muscle.
AuthorsJackson KA,Mi T,Goodell MA
JournalProceedings of the National Academy of Sciences of the United States of America
PubMed ID10588731
We have discovered that cells derived from the skeletal muscle of adult mice contain a remarkable capacity for hematopoietic differentiation. Cells prepared from muscle by enzymatic digestion and 5-day in vitro culture were harvested, and 18 Ã 10(3) cells were introduced into each of six lethally irradiated recipients together with ... More
SHAP potentiates the CD44-mediated leukocyte adhesion to the hyaluronan substratum.
AuthorsZhuo L,Kanamori A,Kannagi R,Itano N,Wu J,Hamaguchi M,Ishiguro N,Kimata K
JournalThe Journal of biological chemistry
PubMed ID16702221
Authors
Journal
PubMed ID16473883
FDF03, a novel inhibitory receptor of the immunoglobulin superfamily, is expressed by human dendritic and myeloid cells.
JournalJournal of immunology (Baltimore, Md. : 1950)
PubMed ID10903717
Identification of kappa opioid receptors in the immune system by indirect immunofluorescence.
AuthorsLawrence DM, el-Hamouly W, Archer S, Leary JF, Bidlack JM
JournalProc Natl Acad Sci U S A
PubMed ID7862634
'A method to visualize the kappa opioid receptor is described that uses a high-affinity fluorescein-conjugated opioid ligand and indirect immunofluorescence with the phycoerythrin fluorophore to amplify the signal. The mouse thymoma cell line R1E/TL8x.1.G1.OUAr.1 (R1EGO), which expresses the kappa 1 but not mu or delta opioid receptors, was used as ... More
Techniques to improve the direct ex vivo detection of low frequency antigen-specific CD8+ T cells with peptide-major histocompatibility complex class I tetramers.
AuthorsChattopadhyay PK, Melenhorst JJ, Ladell K, Gostick E, Scheinberg P, Barrett AJ, Wooldridge L, Roederer M, Sewell AK, Price DA,
JournalCytometry A
PubMed ID18836993
'The ability to quantify and characterize antigen-specific CD8+ T cells irrespective of functional readouts using fluorochrome-conjugated peptide-major histocompatibility complex class I (pMHCI) tetramers in conjunction with flow cytometry has transformed our understanding of cellular immune responses over the past decade. In the case of prevalent CD8+ T cell populations that ... More
'Over the past two decades, the field of biosensors has been developing fast, portable, and convenient detection tools for various molecules of interest, both biological and environmental. Although much attention is paid to the transduction portion of the sensor, the actual bioreceptor that binds the ligand is equally critical. Tight, ... More
Easily reversible desthiobiotin binding to streptavidin, avidin, and other biotin-binding proteins: uses for protein labeling, detection, and isolation.
AuthorsHirsch JD, Eslamizar L, Filanoski BJ, Malekzadeh N, Haugland RP, Beechem JM, Haugland RP
JournalAnal Biochem
PubMed ID12419349
'The high-affinity binding of biotin to avidin, streptavidin, and related proteins has been exploited for decades. However, a disadvantage of the biotin/biotin-binding protein interaction is that it is essentially irreversible under physiological conditions. Desthiobiotin is a biotin analogue that binds less tightly to biotin-binding proteins and is easily displaced by ... More
Development of a ubiquitin transfer assay for high throughput screening by fluorescence resonance energy transfer.
'An assay based on fluorescence resonance energy transfer (FRET) has been developed to screen for ubiquitination inhibitors. The assay measures the transfer of ubiquitin from Ubc4 to HECT protein Rsc 1083. Secondary reagents (streptavidin and antibody to glutathione-S-transferase [GST]), pre-labeled with fluorophores (europium chelate, Eu(3+), and allophycocyanin [APC]), are noncovalently ... More
Detection of surface and cytoplasmic CD4 on blood monocytes from normal and HIV-1 infected individuals.
AuthorsFilion LG, Izaguirre CA, Garber GE, Huebsh L, Aye MT
JournalJ Immunol Methods
PubMed ID1703191
'CD4 on blood monocytes is generally regarded as being found on a subset of blood monocytes. However, our results show that all monocytes are CD4+ but the number of molecules per cell is lower than T cells. We have performed immunofluorescent (flow cytometry, microscopy) analysis of monocytes from normal donors ... More
Flow-cytometric isolation of human antibodies from a nonimmune Saccharomyces cerevisiae surface display library.
'A nonimmune library of 10(9) human antibody scFv fragments has been cloned and expressed on the surface of yeast, and nanomolar-affinity scFvs routinely obtained by magnetic bead screening and flow-cytometric sorting. The yeast library can be amplified 10(10)-fold without measurable loss of clonal diversity, allowing its effectively indefinite expansion. The ... More
Affinity modulation of platelet integrin alphaIIbbeta3 by beta3-endonexin, a selective binding partner of the beta3 integrin cytoplasmic tail.
AuthorsKashiwagi H, Schwartz MA, Eigenthaler M, Davis KA, Ginsberg MH, Shattil SJ
JournalJ Cell Biol
PubMed ID9182673
'Platelet agonists increase the affinity state of integrin alphaIIbbeta3, a prerequisite for fibrinogen binding and platelet aggregation. This process may be triggered by a regulatory molecule(s) that binds to the integrin cytoplasmic tails, causing a structural change in the receptor. beta3-Endonexin is a novel 111-amino acid protein that binds selectively ... More
A novel motor, KIF13A, transports mannose-6-phosphate receptor to plasma membrane through direct interaction with AP-1 complex.
AuthorsNakagawa T, Setou M, Seog D, Ogasawara K, Dohmae N, Takio K, Hirokawa N
JournalCell
PubMed ID11106728
'Intracellular transport mediated by kinesin superfamily proteins (KIFs) is a highly regulated process. The molecular mechanism of KIFs binding to their respective cargoes remains unclear. We report that KIF13A is a novel plus end-directed microtubule-dependent motor protein and associates with beta 1-adaptin, a subunit of the AP-1 adaptor complex. The ... More
Characterization of a CD43/leukosialin-mediated pathway for inducing apoptosis in human T-lymphoblastoid cells.
'The monoclonal antibody (mAb) J393 induces apoptosis in Jurkat T-cells. NH2-terminal amino acid sequence analysis identified the 140-kDa surface antigen for mAb J393 as CD43/leukosialin, the major sialoglycoprotein of leukocytes. While Jurkat cells co-expressed two discrete cell-surface isoforms of CD43, recognized by mAb J393 and mAb G10-2, respectively, only J393/CD43 ... More
The association of CRKII with C3G can be regulated by integrins and defines a novel means to regulate the mitogen-activated protein kinases.
AuthorsBuensuceso CS, O'Toole TE
JournalJ Biol Chem
PubMed ID10777617
'In studies to define mechanisms of ERK activation in Chinese hamster ovary cells, we have observed an inverse correlation between CRKII-C3G complex formation and ERK activity. That is, we were able to coprecipitate the guanine nucleotide exchange factor C3G with the adaptor protein CRKII in lysates from suspended cells that ... More
Rapid generation and flow cytometric analysis of stable GFP-expressing cells.
AuthorsLybarger L, Dempsey D, Franek KJ, Chervenak R
JournalCytometry
PubMed ID8914818
'Expression of green fluorescent protein (GFP) represents a unique method for the fluorescent labeling of viable mammalian cells, with many potential applications. The studies detailed in this report examine the detection of GFP expression in murine cells using flow cytometry. Direct comparison of NIH 3T3 cells transiently expressing GFP or ... More
T cells compete for access to antigen-bearing antigen-presenting cells.
AuthorsKedl RM, Rees WA, Hildeman DA, Schaefer B, Mitchell T, Kappler J, Marrack P
JournalJ Exp Med
PubMed ID11034600
'These studies tested whether antigenic competition between T cells occurs. We generated CD8(+) T cell responses in H-2(b) mice against the dominant ovalbumin epitope SIINFEKL (ova8) and subdominant epitope KRVVFDKL, using either vaccinia virus expressing ovalbumin (VV-ova) or peptide-pulsed dendritic cells. CD8(+) T cell responses were visualized by major histocompatibility ... More
T cell antigen receptor expression by subsets of Ly-2-L3T4- (CD8-CD4-) thymocytes.
AuthorsWilson A, Ewing T, Owens T, Scollay R, Shortman K
JournalJ Immunol
PubMed ID2450128
'The V beta 8-specific mAb F23.1 and KJ16 were used as fluorescent stains to test for TCR expression on the surface of subpopulations of early, CD4-CD8- (L3T4-Ly-2-) thymocytes from adult CBA mice. A surprisingly high proportion (27%) of Ly-2-L3T4- thymocytes were strongly F23.1 and KJ16 positive. No positive cells were ... More
Detection of apoptosis and DNA replication by differential labeling of DNA strand breaks with fluorochromes of different color.
AuthorsLi X, Melamed MR, Darzynkiewicz Z
JournalExp Cell Res
PubMed ID8549670
'Selective DNA strand break induction by photolysis (SBIP) at sites that contain incorporated halogenated nucleotides has been recently proposed as a means of analyzing DNA replication and detecting proliferating cells. The presence of numerous in situ DNA strand breaks is also an inherent feature of apoptotic cells. The aim of ... More
Monitoring cellular responses to Listeria monocytogenes with oligonucleotide arrays.
AuthorsCohen P, Bouaboula M, Bellis M, Baron V, Jbilo O, Poinot-Chazel C, Galiègue S, Hadibi EH, Casellas P
JournalJ Biol Chem
PubMed ID10753925
'Listeria monocytogenes is a pathogenic intracellular microorganism whose infection induces pleiotropic biological changes associated with host cell gene expression regulation. Here we define the gene expression profiles of the human promyelocytic THP1 cell line before and after L. monocytogenes infection. Gene expression was measured on a large scale via oligonucleotide ... More
Steroid and xenobiotic receptor SXR mediates vitamin K2-activated transcription of extracellular matrix-related genes and collagen accumulation in osteoblastic cells.
AuthorsIchikawa T, Horie-Inoue K, Ikeda K, Blumberg B, Inoue S
JournalJ Biol Chem
PubMed ID16606623
'Vitamin K2 is a critical nutrient required for blood coagulation. It also plays a key role in bone homeostasis and is a clinically effective therapeutic agent for osteoporosis. We previously demonstrated that vitamin K2 is a transcriptional regulator of bone marker genes in osteoblastic cells and that it may potentiate ... More
Bacterial transcript imaging by hybridization of total RNA to oligonucleotide arrays.
Authorsde Saizieu A, Certa U, Warrington J, Gray C, Keck W, Mous J
JournalNat Biotechnol
PubMed ID9447592
'We have used high-density oligonucleotide probe arrays (chips) for bacterial transcript imaging. We designed a chip containing probes representing 106 Hemophilus influenzae genes and 100 Streptococcus pneumoniae genes. The apparent lack of polyadenylated transcripts excludes enrichment of mRNA by affinity purification and we thus used total, chemically biotinylated RNA as ... More
Differing roles of inflammation and antigen in T cell proliferation and memory generation.
AuthorsBusch DH, Kerksiek KM, Pamer EG
JournalJ Immunol
PubMed ID10754299
'Recent studies have demonstrated that viral and bacterial infections can induce dramatic in vivo expansion of Ag-specific T lymphocytes. Although presentation of Ag is critical for activation of naive T cells, it is less clear how dependent subsequent in vivo T cell proliferation and memory generation are upon Ag. We ... More
Natural autoreactive B cells in transgenic mice reproduce an apparent paradox to the clonal tolerance theory.
'Naturally occurring autoreactive B cells are thought to be physically eliminated or rendered functionally silent through different mechanisms of tolerance. However, multireactive low affinity natural autoantibody-producing B cells seem to escape these mechanisms in normal adults and could constitute the B cell pool from which pathological autoantibodies can emerge. To ... More
MHC class II tetramers identify peptide-specific human CD4(+) T cells proliferating in response to influenza A antigen.
AuthorsNovak EJ, Liu AW, Nepom GT, Kwok WW
JournalJ Clin Invest
PubMed ID10606632
'Antigen-specific T helper cells present in peripheral blood at very low frequencies are capable of rapid clonal expansion during antigenic challenge. The exquisite specificity of this response provides for activation and expansion of a very select cohort of T cells, a feature we have used to directly identify and quantify ... More
Free cholesterol loading of macrophages induces apoptosis involving the fas pathway.
AuthorsYao PM, Tabas I
JournalJ Biol Chem
PubMed ID10791964
'Macrophage death is an important feature of atherosclerosis, but the cellular mechanism for this process is largely unknown. There is increasing interest in cellular free cholesterol (FC) excess as an inducer of lesional macrophage death because macrophages accumulate large amounts of FC in vivo, and FC loading of macrophages in ... More
Marker-specific sorting of rare cells using dielectrophoresis.
'Current techniques in high-speed cell sorting are limited by the inherent coupling among three competing parameters of performance: throughput, purity, and rare cell recovery. Microfluidics provides an alternate strategy to decouple these parameters through the use of arrayed devices that operate in parallel. To efficiently isolate rare cells from complex ... More
Detection by immunofluorescence of surface molecules present in low copy numbers. High sensitivity staining and calibration of flow cytometer.
AuthorsZola H, Neoh SH, Mantzioris BX, Webster J, Loughnan MS
JournalJ Immunol Methods
PubMed ID1703189
'Receptors for lymphokines and growth factors are present on cell surfaces often at concentrations of 100-500 copies per cell. Although conventional immunofluorescence cannot detect such low levels, cell membrane antigens present at these concentrations can be detected using an optimally set up flow cytometer together with a three-layer immunofluorescence technique, ... More
Cell surface expression of the varicella-zoster virus glycoproteins and Fc receptor.
AuthorsLitwin V, Sandor M, Grose C
JournalVirology
PubMed ID2167554
'Varicella-zoster virus (VZV) specifies the synthesis of viral glycoproteins which are important antigens for induction of the host immune response. In this report the technology of laser-activated flow cytometry has been employed to measure the membrane expression of VZV glycoproteins gpI, gpII, gpIII, and gpIV. By use of biotinylated monoclonal ... More
Detection of low-abundance membrane markers by immunofluorescence--a comparison of alternative high-sensitivity methods and reagents.
'The analysis of membrane molecules using antibodies detected by immunofluorescence staining and flow cytometry is used widely in research and diagnostic immunology. Conventional staining techniques readily detect molecules present at concentrations of around 2000 molecules per cell, but some molecules are expressed and function at much lower abundance. We described ... More
Gene expression profiling of ovarian tissues for determination of molecular pathways reflective of tumorigenesis.
'Ovarian cancer is the fourth leading cause of gynecological cancer death among women in the United States. Early detection is a critical prerequisite to initiating effective cancer therapy. Gene microarray technology and proteomics have provided much of the biomarkers with potential use for diagnosis. However, more research is needed to ... More
Tissue plasminogen activator coexpressed in Chinese hamster ovary cells with alpha(2,6)-sialyltransferase contains NeuAc alpha(2,6)Gal beta(1,4)Glc-N-AcR linkages.
AuthorsMinch SL, Kallio PT, Bailey JE
JournalBiotechnol Prog
PubMed ID7619404
'Genetic alteration of the set of oligosaccharide biosynthesis enzymes expressed in a genetically engineered host cell line is a plausible strategy for manipulating the oligosaccharides on a cloned glycoprotein coexpressed in that cell line. This hypothesis was verified for the particular case of sialylation of recombinant human tissue plasminogen activator ... More
Signal thresholds and modular synergy during expression of costimulatory molecules in B lymphocytes.
AuthorsNatarajan K, Sahoo NC, Rao KV
JournalJ Immunol
PubMed ID11418639
'We analyzed intracellular pathways modulating surface densities of CD80 and CD86 in B cells activated through ligation of the Ag receptor, and the adhesion molecule CD54. Whereas B cell Ag receptor (BCR) cross-linking alone stimulated increased expression of CD86, up-regulation of CD80 required dual stimulation with anti-IgM and anti-CD54. The ... More
PI3P signaling regulates receptor sorting but not transport in the endosomal pathway.
AuthorsPetiot A, Faure J, Stenmark H, Gruenberg J
JournalJ Cell Biol
PubMed ID12975344
'While evidence is accumulating that phosphoinositide signaling plays a crucial role in growth factor and hormone receptor down-regulation, this signaling pathway has also been proposed to regulate endosomal membrane transport and multivesicular endosome biogenesis. Here, we have followed the fate of the down-regulated EGF receptor (EGFR) and bulk transport (fluid ... More
Epstein-Barr virus suspension cell assay using in situ hybridization and flow cytometry.
AuthorsCrouch J, Leitenberg D, Smith BR, Howe JG
JournalCytometry
PubMed ID9298811
'We have developed a procedure for quantitative assay of Epstein-Barr virus (EBV)-infected cells in suspension in either latent or replicative phase using in situ hybridization and flow cytometry. The cells were hybridized with EBV-specific digoxigenin or biotin-labeled oligonucleotide probes, followed by binding to fluorescein-conjugated anti-digoxigenin or phycoerythrin-conjugated streptavidin, respectively. The ... More
Primary structure of an apical protein from Xenopus laevis that participates in amiloride-sensitive sodium channel activity.
'High resistance epithelia express on their apical side an amiloride-sensitive sodium channel that controls sodium reabsorption. A cDNA was found to encode a 1,420-amino acid long polypeptide with no signal sequence, a putative transmembrane segment, and three predicted amphipathic alpha helices. A corresponding 5.2-kb mRNA was detected in Xenopus laevis ... More
Quantitation of microparticles released from coated-platelets.
AuthorsDale GL, Remenyi G, Friese P,
JournalJ Thromb Haemost
PubMed ID16102115
'Dual agonist stimulation of platelets with thrombin and convulxin results in generation of coated-platelets, a sub-population of cells known formerly as COAT-platelets (collagen and thrombin). Coated-platelets retain several procoagulant proteins on their surface and express phosphatidylserine (PS). In this report, we utilize a new methodology to demonstrate that coated-platelets also ... More
Chemical chaperones enhance superantigen and conventional antigen presentation by HLA-DM-deficient as well as HLA-DM-sufficient antigen-presenting cells and enhance IgG2a production in vivo.
AuthorsGhumman B, Bertram EM, Watts TH
JournalJ Immunol
PubMed ID9759841
'Chemical chaperones, first defined in studies of mutant cystic fibrosis transmembrane conductance regulator proteins, are small molecules that act as stabilizers of proteins in their native state and have the ability in some cases to rescue protein-folding mutants within cells. HLA-DM is an MHC II-specific molecular chaperone that facilitates peptide ... More
Flow cytometric analysis of reverse transcription-PCR products: quantification of p21(WAF1/CIP1) and proliferating cell nuclear antigen mRNA.
AuthorsWedemeyer N, Göhde W, Pötter T
JournalClin Chem
PubMed ID10926883
'BACKGROUND: Reverse transcription-PCR (RT-PCR) is a powerful tool in clinical diagnostics for analyzing even small amounts of RNA, but sensitive assays for quantifying the amplification products are time-consuming or expensive. Here we describe a novel flow cytometry-based assay for rapid and sensitive determination of relative amounts of RT-PCR products. METHODS: ... More
Concurrent measurement of the survival of two populations of rabbit platelets labeled with either two PKH lipophilic dyes or two concentrations of biotin.
AuthorsRand ML, Wang H, Mody M, Chu I, Treutiger I, Nguyen A, Packham MA, Freedman J
JournalCytometry
PubMed ID11813201
'BACKGROUND: To avoid radioisotopic labeling and permit comparison of the survival of two platelet populations concurrently in one animal, we compared simultaneous recoveries and survival times of homologous rabbit platelets labeled in vitro with the lipophilic dyes PKH26 (red fluorescing) and PKH67 (green fluorescing) and with two levels of biotin ... More
Optimization of cell permeabilization for multiparametric flow cytometric analysis with lectin staining.
AuthorsVerdier M, Jayat C, Ratinaud MH, Troutaud D
JournalCytometry
PubMed ID10942897
'BACKGROUND: This study was undertaken in mice to develop a reproducible procedure of cell permeabilization, allowing intracellular protein staining by immunofluorescence (i.e., Bcl-2) without losing surface labeling especially for lectins (i.e., B220 and peanut agglutinin [PNA]). This article reports results obtained with different permeabilization protocols. METHODS: Lymphoid cells were extracted ... More
Expression monitoring by hybridization to high-density oligonucleotide arrays.
AuthorsLockhart DJ, Dong H, Byrne MC, Follettie MT, Gallo MV, Chee MS, Mittmann M, Wang C, Kobayashi M, Horton H, Brown EL
JournalNat Biotechnol
PubMed ID9634850
'The human genome encodes approximately 100,000 different genes, and at least partial sequence information for nearly all will be available soon. Sequence information alone, however, is insufficient for a full understanding of gene function, expression, regulation, and splice-site variation. Because cellular processes are governed by the repertoire of expressed genes, ... More
A shift in the phenotype of melan-A-specific CTL identifies melanoma patients with an active tumor-specific immune response.
AuthorsDunbar PR, Smith CL, Chao D, Salio M, Shepherd D, Mirza F, Lipp M, Lanzavecchia A, Sallusto F, Evans A, Russell-Jones R, Harris AL, Cerundolo V
JournalJ Immunol
PubMed ID11086110
'In a significant proportion of melanoma patients, CTL specific for the melan-A(26/7-35) epitope can be detected in peripheral blood using HLA-A2/peptide tetramers. However, the functional capacity of these CTL has been controversial, since although they prove to be effective killers after in vitro expansion, in some patients they have blunted ... More
CD8+ T cell-dependent elimination of dendritic cells in vivo limits the induction of antitumor immunity.
AuthorsHermans IF, Ritchie DS, Yang J, Roberts JM, Ronchese F
JournalJ Immunol
PubMed ID10706699
'The fate of dendritic cells (DC) after they have initiated a T cell immune response is still undefined. We have monitored the migration of DC labeled with a fluorescent tracer and injected s.c. into naive mice or into mice with an ongoing immune response. DC not loaded with Ag were ... More
Diabetes is predicted by the beta cell level of autoantigen.
AuthorsByersdorfer CA, Schweitzer GG, Unanue ER
JournalJ Immunol
PubMed ID16177075
'Two novel transgenic (Tg) strains were created expressing hen egg-white lysozyme (HEL) in a pancreas-specific fashion. RmHP.111 mice had levels of HEL per cell similar to that of the established ILK-3 strain, while RmHP.117 mice had 10-fold lower levels (50,000 molecules per cell). When bred to 3A9 TCR Tg mice, ... More
In vivo identification of glycolipid antigen-specific T cells using fluorescent CD1d tetramers.
AuthorsBenlagha K, Weiss A, Beavis A, Teyton L, Bendelac A
JournalJ Exp Med
PubMed ID10839805
'The CD1 family of major histocompatibility complex (MHC)-like molecules specializes in presenting lipid and glycolipid antigens to alpha/beta T lymphocytes, but little is known about the size of the CD1-restricted T cell population or the frequency of T lymphocytes specific for a given glycolipid antigen. Here, we report the generation ... More
Regulation of the GABA cell phenotype in hippocampus of schizophrenics and bipolars.
'GABAergic dysfunction is present in the hippocampus in schizophrenia (SZ) and bipolar disorder (BD). The trisynaptic pathway was "deconstructed" into various layers of sectors CA3/2 and CA1 and gene expression profiling performed. Network association analysis was used to uncover genes that may be related to regulation of glutamate decarboxylase 67 ... More
Rapid isolation of high-affinity protein binding peptides using bacterial display.
AuthorsBessette PH, Rice JJ, Daugherty PS
JournalProtein Eng Des Sel
PubMed ID15531628
'A robust bacterial display methodology was developed that allows the rapid isolation of peptides that bind to arbitrarily selected targets with high affinity. To demonstrate the utility of this approach, a large library (5 x 10(10) clones) was constructed composed of random 15-mer peptide insertions constrained within a flexible, surface ... More
Thiol levels in CD134-defined subsets of rat T lymphocytes: possible implications for HgCl2-induced immune dysregulation.
AuthorsRoos A, Claessen N, Schilder-Tol EJ, Chand MA, Weening JJ, Aten J
JournalBiochem Biophys Res Commun
PubMed ID9388500
'CD134 (OX40), a member of the tumour necrosis factor receptor family, is expressed on activated T cells and mediates T and B cell costimulation. Its expression is increased after exposure to the thiol-binding compound HgCl2 in BN rats, but not in Lewis rats, in association with induction of a T ... More
Cryptomonad algal phycobiliproteins as fluorochromes for extracellular and intracellular antigen detection by flow cytometry.
AuthorsTelford WG, Moss MW, Morseman JP, Allnutt FC
JournalCytometry
PubMed ID11309804
'BACKGROUND: Phycobiliproteins play an important role in fluorescent labeling, particularly for flow cytometry. The spectral properties of R-phycoerythrin (R-PE) and allophycocyanin (APC) have made them the dominant reagents in this class of fluorochromes. In this study, we evaluate a lesser-known but potentially important series of low-molecular weight cryptomonad-derived phycobiliproteins (commercially ... More
Inducible IL-2 production by dendritic cells revealed by global gene expression analysis.
AuthorsGranucci F, Vizzardelli C, Pavelka N, Feau S, Persico M, Virzi E, Rescigno M, Moro G, Ricciardi-Castagnoli P
JournalNat Immunol
PubMed ID11526406
'Dendritic cells (DCs) are strong activators of primary T cell responses. Their priming ability is acquired upon encounter with maturation stimuli. To identify the genes that are differentially expressed upon maturation induced by exposure to Gram-negative bacteria, a kinetic study of DC gene expression was done with microarrays representing 11,000 ... More
Parallel detection of antigen-specific T-cell responses by multidimensional encoding of MHC multimers.
AuthorsHadrup SR, Bakker AH, Shu CJ, Andersen RS, van Veluw J, Hombrink P, Castermans E, Thor Straten P, Blank C, Haanen JB, Heemskerk MH, Schumacher TN,
JournalNat Methods
PubMed ID19543285
'The use of fluorescently labeled major histocompatibility complex multimers has become an essential technique for analyzing disease- and therapy-induced T-cell immunity. Whereas classical major histocompatibility complex multimer analyses are well-suited for the detection of immune responses to a few epitopes, limitations on human-subject sample size preclude a comprehensive analysis of ... More
Nonproliferating bystander CD4+ T cells lacking activation markers support HIV replication during immune activation.
AuthorsScales D, Ni H, Shaheen F, Capodici J, Cannon G, Weissman D
JournalJ Immunol
PubMed ID11342670
'HIV replicates primarily in lymphoid tissue and immune activation is a major stimulus in vivo. To determine the cells responsible for HIV replication during Ag-driven T cell activation, we used a novel in vitro model employing dendritic cell presentation of superantigen to CD4(+) T cells. Dendritic cells and CD4(+) T ... More
Relationship of DNA ploidy to chemoresistance of tumors as measured by in vitro tests.
AuthorsVolm M, Efferth T
JournalCytometry
PubMed ID1971209
'To examine whether patients with aneuploid tumors might derive more benefit from chemotherapy than would patients with diploid tumors, predictive tests for determining resistance in human tumors were carried out and the test results compared with the DNA ploidy of the corresponding tumors. Multidrug-resistance in 15 kidney carcinomas grown as ... More
Apoptosis of T cells mediated by galectin-1.
AuthorsPerillo NL, Pace KE, Seilhamer JJ, Baum LG
JournalNature
PubMed ID7501023
'Galectin-1, a member of the family of beta-galactoside binding proteins, has growth regulatory and immunomodulatory activities. We report here that galectin-1, expressed by stromal cells in human thymus and lymph nodes, is present at sites of cell death by apoptosis during normal T-cell development and maturation. Galectin-1 induced apoptosis of ... More
CD4+CD8+ thymocytes develop into CD4 or CD8 single-positive cells in athymic nude mice.
AuthorsTamauchi H, Tamaoki N, Habu S
JournalEur J Immunol
PubMed ID3144457
'A differentiation pathway from CD4+CD8+ cells to CD4+CD8- or CD4-CD8+ cells was investigated in athymic nude mice. Using fluorescence-activated cell sorter, CD4+CD8+ cells were sorted out from AKR thymocytes (H-2k, Thy-1.1) stained with two monoclonal antibodies against CD4 and CD8 (anti-L3T4 and anti-Ly-2). These CD4+CD8+ AKR thymocytes were injected i.v. ... More
Identification of transcriptional targets of HOXA5.
AuthorsChen H, Rubin E, Zhang H, Chung S, Jie CC, Garrett E, Biswal S, Sukumar S
JournalJ Biol Chem
PubMed ID15757903
'The homeobox gene HOXA5 encodes a transcription factor that has been shown to play important roles in embryogenesis, hematopoiesis, and tumorigenesis. In order to decipher downstream signaling pathways of HOXA5, we utilized oligonucleotide microarray analysis to identify genes that are differentially expressed in HOXA5-induced cells compared with uninduced cells. Comparative ... More
Variable immunodominance hierarchies for H2-M3-restricted N-formyl peptides following bacterial infection.
AuthorsKerksiek KM, Busch DH, Pamer EG
JournalJ Immunol
PubMed ID11145694
'H2-M3-restricted presentation of N-formyl methionine (f-Met) peptides to CD8(+) T cells provides a mechanism for selective recognition of bacterial infection. In this report we demonstrate that Listeria monocytogenes infection induces distinct CD8(+) T cell populations specific for each of the known Listeria-derived formyl methionine peptides presented by M3. The sum ... More
Expression profiling of pancreatic beta cells: glucose regulation of secretory and metabolic pathway genes.
AuthorsWebb GC, Akbar MS, Zhao C, Steiner DF
JournalProc Natl Acad Sci U S A
PubMed ID10811900
'Pancreatic beta cells respond to changes in blood glucose by secreting insulin and increasing insulin synthesis. To identify genes used in these responses, we have carried out expression profiling of beta cells exposed to high (25 mM) or low (5.5 mM) glucose by using oligonucleotide microarrays. Functional clustering of genes ... More
Thrombin responses in human endothelial cells. Contributions from receptors other than PAR1 include the transactivation of PAR2 by thrombin-cleaved PAR1.
AuthorsO'Brien PJ, Prevost N, Molino M, Hollinger MK, Woolkalis MJ, Woulfe DS, Brass LF
JournalJ Biol Chem
PubMed ID10788464
'The recent identification of two new thrombin receptors, PAR3 and PAR4, led us to re-examine the basis for endothelial cell responses to thrombin. Human umbilical vein endothelial cells (HUVEC) are known to express PAR1 and the trypsin/tryptase receptor, PAR2. Northern blots detected both of those receptors and, to a lesser ... More
Critical role for CD8 in binding of MHC tetramers to TCR: CD8 antibodies block specific binding of human tumor-specific MHC-peptide tetramers to TCR.
AuthorsDenkberg G, Cohen CJ, Reiter Y
JournalJ Immunol
PubMed ID11418659
'There are conflicting opinions about the role that the T cell coreceptors CD4 and CD8 play in TCR binding and activation. Recent evidence from transgenic mouse models suggests that CD8 plays a critical role in TCR binding and activation by peptide-MHC complex multimers (tetramers). Here we show with a human ... More
Green- and red-fluorescent nanospheres for the detection of cell surface receptors by flow cytometry.
AuthorsBhalgat MK, Haugland RP, Pollack JS, Swan S, Haugland RP
JournalJ Immunol Methods
PubMed ID9831388
'Fluorescent probes serve as sensitive tools for obtaining structural and functional information in cellular systems. In spite of the high sensitivity provided by fluorescent reagents, cell surface receptors expressed in low numbers often escape detection with commonly used fluorescent probes. R-Phycoerythrin (R-PE), a molecule with a very high quantum yield, ... More
Preparing fluorescent probes for microarray studies.
AuthorsXiang CC, Brownstein MJ
JournalMethods Mol Biol
PubMed ID12710665
Immunostimulatory effects of a plasmid expressing CD40 ligand (CD154) on gene immunization.
AuthorsMendoza RB, Cantwell MJ, Kipps TJ
JournalJ Immunol
PubMed ID9550372
'Interaction of CD40 with its ligand (CD154) can induce CD40-bearing APCs to express immune stimulatory accessory molecules that facilitate immune recognition. We evaluated whether a plasmid vector encoding CD154 (pCD40L) could influence the immune response to a transgene protein encoded by coinjected plasmid DNA. We found that coinjection of pCD40L ... More
Quantitation of HIV-1-specific cytotoxic T lymphocytes and plasma load of viral RNA.
AuthorsOgg GS, Jin X, Bonhoeffer S, Dunbar PR, Nowak MA, Monard S, Segal JP, Cao Y, Rowland-Jones SL, Cerundolo V, Hurley A, Markowitz M, Ho DD, Nixon DF, McMichael AJ
JournalScience
PubMed ID9516110
'Although cytotoxic T lymphocytes (CTLs) are thought to be involved in the control of human immunodeficiency virus-type 1 (HIV-1) infection, it has not been possible to demonstrate a direct relation between CTL activity and plasma RNA viral load. Human leukocyte antigen-peptide tetrameric complexes offer a specific means to directly quantitate ... More
Autocrine and paracrine apoptosis are mediated by differential regulation of Fas ligand activity in two distinct Jurkat T cell populations.
AuthorsSu X, Cheng J, Liu W, Liu C, Wang Z, Yang P, Zhou T, Mountz JD
JournalJ Immunol
PubMed ID9605126
'Fas ligand (FasL) produced by activated T cells mediates autocrine-induced apoptosis to limit T cell expansion. To investigate the regulation of FasL activity, Jurkat cells were stably transfected with a 2.3-kb fragment of human FasL promoter that controlled the expression of a GFP reporter gene. Two populations of Jurkat cells ... More
Induced multidrug resistance in murine leukemia L1210 and associated changes in a surface-membrane glycoprotein.
AuthorsVolm M, Bak M, Efferth T, Mattern J
JournalJ Cancer Res Clin Oncol
PubMed ID2563999
'The aim of this study was to find out whether only resistant cells of the "multidrug-resistant" phenotype show the described changes of plasma membrane glycoprotein (170 kDa) or whether resistant cells that do not express this phenotype reveal corresponding results. Doxorubicin-resistant (L1210dox) and daunorubicin-resistant L1210 ascites tumor cells (L1210dnr) (multidrug-resistant ... More
Diverse signaling pathways activated by growth factor receptors induce broadly overlapping, rather than independent, sets of genes.
AuthorsFambrough D, McClure K, Kazlauskas A, Lander ES
JournalCell
PubMed ID10380925
'We sought to explore the relationship between receptor tyrosine kinase (RTK) activated signaling pathways and the transcriptional induction of immediate early genes (IEGs). Using global expression monitoring, we identified 66 fibroblast IEGs induced by platelet-derived growth factor beta receptor (PDGFRbeta) signaling. Mutant receptors lacking binding sites for activation of the ... More
Profiling changes in gene expression during differentiation and maturation of monocyte-derived dendritic cells using both oligonucleotide microarrays and proteomics.
AuthorsLe Naour F, Hohenkirk L, Grolleau A, Misek DE, Lescure P, Geiger JD, Hanash S, Beretta L
JournalJ Biol Chem
PubMed ID11279020
'Dendritic cells (DCs) are antigen-presenting cells that play a major role in initiating primary immune responses. We have utilized two independent approaches, DNA microarrays and proteomics, to analyze the expression profile of human CD14(+) blood monocytes and their derived DCs. Analysis of gene expression changes at the RNA level using ... More
Chemosensitivity prediction by transcriptional profiling.
AuthorsStaunton JE, Slonim DK, Coller HA, Tamayo P, Angelo MJ, Park J, Scherf U, Lee JK, Reinhold WO, Weinstein JN, Mesirov JP, Lander ES, Golub TR
JournalProc Natl Acad Sci U S A
PubMed ID11553813
'In an effort to develop a genomics-based approach to the prediction of drug response, we have developed an algorithm for classification of cell line chemosensitivity based on gene expression profiles alone. Using oligonucleotide microarrays, the expression levels of 6,817 genes were measured in a panel of 60 human cancer cell ... More
Global response of Saccharomyces cerevisiae to an alkylating agent.
AuthorsJelinsky SA, Samson LD
JournalProc Natl Acad Sci U S A
PubMed ID9990050
'DNA chip technology enables simultaneous examination of how approximately 6,200 Saccharomyces cerevisiae gene transcript levels, representing the entire genome, respond to environmental change. By using chips bearing oligonucleotide arrays, we show that, after exposure to the alkylating agent methyl methanesulfonate, approximately 325 gene transcript levels are increased and approximately 76 ... More
Gene expression patterns in dendritic cells infected with measles virus compared with other pathogens.
AuthorsZilliox MJ, Parmigiani G, Griffin DE
JournalProc Natl Acad Sci U S A
PubMed ID16492729
'Gene expression patterns supply insight into complex biological networks that provide the organization in which viruses and host cells interact. Measles virus (MV) is an important human pathogen that induces transient immunosuppression followed by life-long immunity in infected individuals. Dendritic cells (DCs) are potent antigen-presenting cells that initiate the immune ... More
Isolation and functional properties of murine hematopoietic stem cells that are replicating in vivo.
'Hematopoietic stem cells (HSC) are multipotent cells that reside in the bone marrow and replenish all adult hematopoietic lineages throughout the lifetime of the animal. While experimenting with staining of murine bone marrow cells with the vital dye, Hoechst 33342, we discovered that display of Hoechst fluorescence simultaneously at two ... More
Cell division is not a "clock" measuring acquisition of competence to produce IFN-gamma or IL-4.
AuthorsBen-Sasson SZ, Gerstel R, Hu-Li J, Paul WE
JournalJ Immunol
PubMed ID11123283
'Naive CD4 T cells acquire the potential to produce IFN-gamma and IL-4 by culture in the presence of their cognate Ag, APC, and appropriate cytokines. In this study, we show that commitment to IFN-gamma production on the part of rigorously purified naive CD4 T cells can occur without cell division. ... More
Inhibition of calcium mobilization is an early event in opiate-induced immunosuppression.
AuthorsSei Y, McIntyre T, Fride E, Yoshimoto K, Skolnick P, Arora PK
JournalFASEB J
PubMed ID2022315
'Morphine administered as a subcutaneous implant inhibits the initial increase in cytoplasmic free-calcium [Ca2+]i induced by mitogens in mouse splenocytes. This effect was not reproduced by incubation of splenocytes with morphine (10(-8)-10(-4) M). Analysis of splenocyte subpopulations demonstrates that this effect was manifest in both B and T cells. However, ... More
Phycobiliprotein-Fab conjugates as probes for single particle fluorescence imaging.
AuthorsTriantafilou K, Triantafilou M, Wilson KM
JournalCytometry
PubMed ID11042621
'BACKGROUND AND METHODS: Single particle fluorescence imaging (SPFI) is a recently developed method that has provided a powerful approach to observing receptor movement and associations at high spatial resolution. It provides a noninvasive alternative to the existing biochemical techniques. It can also quantify and resolve molecular interactions at the cell ... More
Frequency of class I HLA-restricted anti-HIV CD8+ T cells in individuals receiving highly active antiretroviral therapy (HAART).
AuthorsGray CM, Lawrence J, Schapiro JM, Altman JD, Winters MA, Crompton M, Loi M, Kundu SK, Davis MM, Merigan TC
JournalJ Immunol
PubMed ID9973442
'Peptide/MHC tetrameric complexes were used to enumerate the frequency of HLA class I-restricted epitope-specific CD8+ T cells in 18 HLA-A*0201 HIV type 1-infected asymptomatic patients. HLA-A*0201 molecules were complexed to HIV Gag p17 (amino acids 77-85) and reverse transcriptase (amino acids 464-472) peptides, biotinylated, and bound to streptavidin-phycoerythrin to form ... More
A dominant function of IKK/NF-kappaB signaling in global lipopolysaccharide-induced gene expression.
AuthorsCarayol N, Chen J, Yang F, Jin T, Jin L, States D, Wang CY
JournalJ Biol Chem
PubMed ID16914552
'Porphyromonas gingivalis is an etiologic pathogen of periodontitis that is one of the most common inflammatory diseases. Recently, we found that P. gingivalis LPS activated the transcription factor nuclear factor-kappaB (NF-kappaB) through the IkappaB kinase complex (IKK). NF-kappaB is a transcription factor that controls inflammation and host responses. In this ... More
Facile generation of heat-stable antiviral and antitoxin single domain antibodies from a semisynthetic llama library.
AuthorsGoldman ER, Anderson GP, Liu JL, Delehanty JB, Sherwood LJ, Osborn LE, Cummins LB, Hayhurst A
JournalAnal Chem
PubMed ID17165813
'Llamas possess a class of unconventional immunoglobulins that have only heavy chains; unpaired heavy variable domains are responsible for antigen binding. These domains have previously been cloned and expressed as single domain antibodies (sdAbs); they comprise the smallest known antigen binding fragments. SdAbs have been shown to bind antigens at ... More
Up-regulation of interleukin 4/B-cell stimulatory factor 1 receptor expression.
AuthorsOhara J, Paul WE
JournalProc Natl Acad Sci U S A
PubMed ID3263648
'The expression of interleukin 4 (IL-4) receptors on resting T and B lymphocytes was enhanced 4- to 8-fold by IL-4 stimulation of these cells. Other agents such as lipopolysaccharide and anti-IgM for B cells and concanavalin A for T cells also caused increased IL-4 receptor expression, although to a somewhat ... More
Simultaneous detection of antibodies to cytomegalovirus and herpes simplex virus by using flow cytometry and a microsphere-based fluorescence immunoassay.
AuthorsMcHugh TM, Miner RC, Logan LH, Stites DP
JournalJ Clin Microbiol
PubMed ID2846633
'A sensitive assay for the simultaneous detection of anti-cytomegalovirus and anti-herpes simplex virus antibodies was developed. Two different sizes of polystyrene microspheres were coated with purified viral antigens. Human antiviral antibodies were detected with a biotin-streptavidin amplification procedure with phycoerythrin as the fluorescent label. Microsphere-associated fluorescence was quantitated with a ... More
Quantitative analysis of calcium (Ca2+) mobilization after stimulation with mitogens or anti-CD3 antibodies. Simultaneous fluo-3 and immunofluorescence flow cytometry.
AuthorsSei Y, Arora PK
JournalJ Immunol Methods
PubMed ID1672879
'A method is described for flow microfluorometric analysis of calcium mobilization in immune cells. This method is based on dual-color analysis of heterogeneous cell populations using fluo-3 and phycoerythrin (PE)-conjugated monoclonal antibody (mAb). This technique allows the detection of fluo-3 fluorescence as a measure of an increase in cytoplasmic free ... More
Development of a dual label fluorescence technique that can be utilized to elucidate the mechanism of action of monoclonal antibody-drug conjugates.
AuthorsStarling JJ, Hinson NA, Marder P, Maciak RS, Laguzza BC, Corvalan JR, Smith W
JournalCancer Res
PubMed ID3048656
'A method is described that allows the simultaneous visualization and relative assessment of both the antibody and drug components of monoclonal antibody-drug conjugates at the target cell membrane. The antibody is detected by a fluorescein-conjugated anti-mouse immunoglobulin serum while the drug is visualized by rhodamine avidin or phycoerythrin-streptavidin binding to ... More
A multiplex approach to isotyping antigen-specific antibodies using biotinylated antigen/streptavidin-phycoerythrin.
AuthorsMcCutcheon K,
JournalMethods Mol Biol
PubMed ID18287660
'Analytical methods characterizing the immunogenicity of antigens are useful for monitoring, characterizing and predicting antibody responses to therapeutic biologics or vaccines. Distinct Luminex microspheres coupled with protein G, anti-human immunoglobulin (Ig)A, anti-human IgM and anti-human IgE were developed for the simultaneous capture of total IgG, IgA, IgM and IgE (IgGAME) ... More
Sequence requirements of the GPNG beta-turn of the Ecballium elaterium trypsin inhibitor II explored by combinatorial library screening.
AuthorsWentzel A, Christmann A, Krätzner R, Kolmar H
JournalJ Biol Chem
PubMed ID10409654
'The Ecballium elaterium trypsin inhibitor II (EETI-II) contains 28 amino acids and three disulfides forming a cystine knot. Reduced EETI-II refolds spontaneously and quantitatively in vitro and regains its native structure. Due to its high propensity to form a reverse turn, the GPNG sequence of segment 22-25 comprising a beta-turn ... More
Binding of anticoagulant vitamin K-dependent protein S to platelet-derived microparticles.
AuthorsDahlbäck B, Wiedmer T, Sims PJ
JournalBiochemistry
PubMed ID1463747
'Vitamin K-dependent protein S is an anticoagulant plasma protein serving as cofactor to activated protein C in degradation of coagulation factors Va and VIIIa on membrane surfaces. In addition, it forms a noncovalent complex with complement regulatory protein C4b-binding protein (C4BP), a reaction which inhibits its anticoagulant function. Both forms ... More
Quantification of IgG anti-D bound to D-positive red cells infused into D-negative subjects after intramuscular injection of monoclonal anti-D.
AuthorsKumpel BM, Judson PA
JournalTransfus Med
PubMed ID7655572
'A flow-cytometric method was developed to determine the number of molecules of IgG bound to D-positive red blood cells (RBC) when sensitized with low plasma concentrations of IgG anti-D in the presence of an excess of D-negative RBC. D-positive RBC were infused into 12 D-negative male volunteers 2 days after ... More
Cutting edge: germinal centers can be induced in the absence of T cells.
AuthorsLentz VM, Manser T
JournalJ Immunol
PubMed ID11418626
'Immunization of mice containing mutations that inactivate the TCR Cbeta and Cdelta genes with the T cell-independent (TI) type 2 Ag (4-hydroxy-3-nitrophenyl)acetyl-Ficoll induces clusters of peanut agglutinin-binding B cells in the spleen. These clusters are histologically indistinguishable from germinal centers (GCs) typical of T cell-dependent immune responses. They are located ... More
Detection of activated platelets in whole blood using activation-dependent monoclonal antibodies and flow cytometry.
AuthorsShattil SJ, Cunningham M, Hoxie JA
JournalBlood
PubMed ID3297204
'Platelets may become activated in a number of clinical disorders and participate in thrombus formation. We developed a direct test for activated platelets in whole blood using flow cytometry. Whole blood was incubated with either biotin-PAC1, a monoclonal antibody specific for the fibrinogen receptor on activated platelets, or biotin-S12, an ... More
Expression of T cell antigen receptor heterodimers in a lipid-linked form.
AuthorsLin AY, Devaux B, Green A, Sagerström C, Elliott JF, Davis MM
JournalScience
PubMed ID1696397
'The interaction of the T cell receptor for antigen (TCR) with its antigen-major histocompatibility complex ligand is difficult to study because both are cell surface multimers. The TCR consists of two chains (alpha and beta) that are complexed to the five or more nonpolymorphic CD3 polypeptides. A soluble form of ... More
Ultrasensitive detection of biomolecules with fluorescent dye-doped nanoparticles.
AuthorsLian W, Litherland SA, Badrane H, Tan W, Wu D, Baker HV, Gulig PA, Lim DV, Jin S
JournalAnal Biochem
PubMed ID15464962
'Fluorescent-labeled molecules have been used extensively for a wide range of applications in biological detection and diagnosis. A new form of highly luminescent and photostable nanoparticles was generated by doping the fluorescent dye tris(2''2-bipyridyl)dichlororuthenium(II)hexahydrate (Rubpy) inside silica material. Because thousands of fluorescent dye molecules are encapsulated in the silica matrix ... More
Identification of opioid receptors in the immune system using a novel combination of selective opioid ligands and indirect phycoerythrin immunofluorescence.
AuthorsLawrence DM, Archer S, Bidlack JM
JournalAdv Exp Med Biol
PubMed ID7668148
The arrival of HLA class II tetramers.
AuthorsMcMichael AJ, Kelleher A
JournalJ Clin Invest
PubMed ID10606619
"DAKLI": a multipurpose ligand with high affinity and selectivity for dynorphin (kappa opioid) binding sites.
AuthorsGoldstein A, Nestor JJ, Naidu A, Newman SR
JournalProc Natl Acad Sci U S A
PubMed ID2902630
We describe a synthetic ligand, "DAKLI" (Dynorphin A-analogue Kappa LIgand), related to the opioid peptide dynorphin A. A single reactive amino group at the extended carboxyl terminus permits various reporter groups to be attached, such as 125I-labeled Bolton-Hunter reagent, fluorescein isothiocyanate, or biotin. These derivatives have high affinity and selectivity ... More
Suppression of endogenous avidin-binding activity in tissues and its relevance to biotin-avidin detection systems.
AuthorsWood GS, Warnke R
JournalJ Histochem Cytochem
PubMed ID7028859
As biotin-avidin systems continue to be developed for applications involving single cells, cell suspensions, and especially tissue sections, the need arises for a method of blocking endogenous avidin-binding activity. One such method is described and its proposed mechanism is discussed. Utilizing this method, endogenous avidin-binding activity was detected and suppressed ... More
Linear differentiation of cytotoxic effectors into memory T lymphocytes.
AuthorsOpferman JT, Ober BT, Ashton-Rickardt PG
JournalScience
PubMed ID10073942
A central question in immunology is the origin of long-lived T cell memory that confers protection against recurrent infection. The differentiation of naïve T cell receptor transgenic CD8+ cells into effector cytotoxic T lymphocytes (CTLs) and memory CD8+ cells was studied. Memory CD8+ cells that were generated after strong antigenic ... More
Rapid production of quasi-stable antibody-phycoerythrin conjugates for use in flow cytometry.
AuthorsDale GL
JournalCytometry
PubMed ID9845444
A simple system is described for producing quasi-stable complexes between biotinylated antibodies and fluorochrome-labeled streptavidin. Optimal complexes were produced by mixing biotinylated antibody and phycoerythrin-streptavidin together at a molar ratio of 1:1 for 2 min, followed by the addition of soluble biotin to halt oligomerization. These complexes, which are stable ... More
Essential role of MD-2 in LPS responsiveness and TLR4 distribution.
AuthorsNagai Y, Akashi S, Nagafuku M, Ogata M, Iwakura Y, Akira S, Kitamura T, Kosugi A, Kimoto M, Miyake K
JournalNat Immunol
PubMed ID12055629
Toll-like receptor 4 (TLR4) mediates lipopolysaccharide (LPS) signaling in a variety of cell types. MD-2 is associated with the extracellular domain of TLR4 and augments TLR4-dependent LPS responses in vitro. We show here that MD-2(-/-) mice do not respond to LPS, do survive endotoxic shock but are susceptible to Salmonella ... More
Activation of NK cells and T cells by NKG2D, a receptor for stress-inducible MICA.
AuthorsBauer S, Groh V, Wu J, Steinle A, Phillips JH, Lanier LL, Spies T
JournalScience
PubMed ID10426993
Stress-inducible MICA, a distant homolog of major histocompatibility complex (MHC) class I, functions as an antigen for gammadelta T cells and is frequently expressed in epithelial tumors. A receptor for MICA was detected on most gammadelta T cells, CD8+ alphabeta T cells, and natural killer (NK) cells and was identified ... More
Oral cancer in vivo gene expression profiling assisted by laser capture microdissection and microarray analysis.
AuthorsAlevizos I, Mahadevappa M, Zhang X, Ohyama H, Kohno Y, Posner M, Gallagher GT, Varvares M, Cohen D, Kim D, Kent R, Donoff RB, Todd R, Yung CM, Warrington JA, Wong DT
JournalOncogene
PubMed ID11593428
Large scale gene expression profiling was carried out on laser capture microdissected (LCM) tumor and normal oral epithelial cells and analysed on high-density oligonucleotide microarrays. About 600 genes were found to be oral cancer associated. These oral cancer associated genes include oncogenes, tumor suppressors, transcription factors, xenobiotic enzymes, metastatic proteins, ... More