Detection of viable Listeria monocytogenes with a 5' nuclease PCR assay.
AuthorsNorton DM,Batt CA
JournalApplied and environmental microbiology
PubMed ID10224010
A 5â² nuclease assay has been developed to detect viable Listeria monocytogenes. The assay targets the hemolysin A (hlyA) transcript, which is found only in L. monocytogenes. The single-tube, reverse transcriptase (RT), fluorogenic probe-based assay was formatted by using Tth DNA polymerase whose activity was modulated by using the manganese-chelating ... More
MHC class I expression on tumor targets inhibits natural killer cell-mediated cytotoxicity without interfering with target recognition.
AuthorsKaufman DS, Schoon RA, Leibson PJ
JournalJ Immunol
PubMed ID8432986
'NK cell-mediated killing is inversely proportional to the amount of MHC class I expression on certain tumor targets. Two hypotheses have been proposed to explain why class I-bearing targets are more resistant to NK cell-mediated lysis: 1) the presence of class I prevents NK cell recognition of a triggering molecule ... More
A habitat for psychrophiles in deep Antarctic ice.
AuthorsPrice PB
JournalProc Natl Acad Sci U S A
PubMed ID10655516
'Microbes, some of which may be viable, have been found in ice cores drilled at Vostok Station at depths down to approximately 3,600 m, close to the surface of the huge subglacial Lake Vostok. Two types of ice have been found. The upper 3,500 m comprises glacial ice containing traces ... More
Glycolipid-anchored form of CD4 increases intercellular adhesion but is unable to enhance T cell activation.
AuthorsSleckman BP, Rosenstein Y, Igras VE, Greenstein JL, Burakoff SJ
JournalJ Immunol
PubMed ID1677022
'CD4 functions to enhance T cell activation by increasing intercellular adhesion and/or by transduction of an intracellular signal. To study the role of human CD4 in T cell activation we have used a murine T cell hybridoma, By 155.16, which produces IL-2 when stimulated by HLA-DR-bearing cells. Previously, we have ... More
Examination of sodium/glucose cotransport by using a visible glucose analogue.
AuthorsPeerce BE
JournalBiochemistry
PubMed ID2021610
'The glucose derivative, 2,2,6,6-tetramethylpiperidine-1-oxylglucose (TEMPO-glucose) was synthesized and examined for its ability to substitute for glucose as a substrate for the intestinal brush border membrane Na+/glucose cotransporter. TEMPO-glucose inhibited Na(+)-dependent phlorizin binding with an apparent KI of 18 microM and Na(+)-dependent glucose uptake with an apparent KI of 70 microM. ... More
AuthorsGeijtenbeek TB, Krooshoop DJ, Bleijs DA, van Vliet SJ, van Duijnhoven GC, Grabovsky V, Alon R, Figdor CG, van Kooyk Y
JournalNat Immunol
PubMed ID11017109
'Dendritic cells (DCs) are recruited from blood into tissues to patrol for foreign antigens. After antigen uptake and processing, DCs migrate to the secondary lymphoid organs to initiate immune responses. We now show that DC-SIGN, a DC-specific C-type lectin, supports tethering and rolling of DC-SIGN-positive cells on the vascular ligand ... More
Continuous cell activation is necessary for stable interaction of complement receptor type 3 with its counter-structure in the aggregation response of human neutrophils.
AuthorsKuypers TW, Koenderman L, Weening RS, Verhoeven AJ, Roos D
JournalEur J Immunol
PubMed ID2180724
'Human neutrophils aggregate after stimulation with various stimuli; this response is completely absent in neutrophils from patients with leukocyte adhesion deficiency (LAD). To investigate the cellular requirements of this process a method was used in which neutrophils are separately loaded with hydroethidine (HE) and sulfofluorescein (SFDA), to give them either ... More
Tissue concentration and urinary excretion pattern of sulfofluorescein by the rat kidney.
AuthorsAmmer U, Natochin Y, Ullrich KJ
JournalJ Am Soc Nephrol
PubMed ID8490119
'In order to find a fluorescein analog that is excreted in a way similar to p-aminohippurate (PAH) and is suitable to register excretion into the urine and also to monitor continuously the concentration within cortical tissue, the interference of fluorescein, sulfofluorescein (SF), and fluorescein-5(6)-sulfonate with the contraluminal transport systems of ... More
Adhesion of T and B lymphocytes to extracellular matrix and endothelial cells can be regulated through the beta subunit of VLA.
Authorsvan de Wiel-van Kemenade E, van Kooyk Y, de Boer AJ, Huijbens RJ, Weder P, van de Kasteele W, Melief CJ, Figdor CG
JournalJ Cell Biol
PubMed ID1560035
'Investigating the regulation of very late antigen (VLA)-mediated functions, we found that TS2/16, a mAb directed against the beta chain of the VLA group of integrins, can induce binding of resting peripheral blood lymphocytes, cloned T lymphocytes, and Epstein Barr virus-transformed B cells to extracellular matrix components, fibronectin, laminin, and ... More
Modulation of K+ conductance by intracellular pH in pancreatic beta-cells.
AuthorsRosario LM, Rojas E
JournalFEBS Lett
PubMed ID2422060
Measurements of the effects of NH3/NH4+ on glucose-induced electrical activity in beta-cells from microdissected mouse islets of Langerhans and on intracellular pH in single collagenase-isolated islets pre-loaded with a fluorescent pH probe were performed and are reported here. Application of NH3/NH4+ (15 mM) in the presence of glucose (11 mM) ... More
Monitoring of neutrophil priming in whole blood by antibodies isolated from a synthetic phage antibody library.
AuthorsKoenderman L, Kanters D, Maesen B, Raaijmakers J, Lammers JW, de Kruif J, Logtenberg T
JournalJ Leukoc Biol
PubMed ID10914490
Neutrophil activation is a multistep process. In vitro activation of neutrophils with semiphysiological activators is optimal only after preactivation or priming with cytokines, chemotaxins, and/or bacterial products. Until now, no antibodies have been developed that can distinguish between resting and (cytokine) primed neutrophils with a sufficient dynamic range necessary for ... More
Shedding of ICAM-1 from human melanoma cell lines induced by IFN-gamma and tumor necrosis factor-alpha. Functional consequences on cell-mediated cytotoxicity.
AuthorsBecker JC, Dummer R, Hartmann AA, Burg G, Schmidt RE
JournalJ Immunol
PubMed ID1684377
ICAM-1-mediated cell-cell adhesion is essential for various immunologic functions, including non-MHC-restricted cytotoxicity. The present study was designed to establish whether shedding of ICAM-1 from melanoma cells occurred and to characterize the effects of soluble ICAM-1 on some cell adhesion-dependent functions. The shed soluble ICAM-1 molecule was detected and quantified by ... More
Cross-linking of sialophorin (CD43) induces neutrophil aggregation in a CD18-dependent and a CD18-independent way.
AuthorsKuijpers TW, Hoogerwerf M, Kuijpers KC, Schwartz BR, Harlan JM
JournalJ Immunol
PubMed ID1353101
Normal human neutrophils bound an as yet unclustered mAb designated BS-1. The Ag immunoprecipitated with BS-1 was blotted by CD43 mAb (and vice versa), and is therefore identical to the large sialoglycoprotein. The CD43 Ag expression on the neutrophil surface is decreased upon neutrophil activation with the chemoattractant FMLP or ... More
Molecular basis for the homophilic activated leukocyte cell adhesion molecule (ALCAM)-ALCAM interaction.
Activated leukocyte cell adhesion molecule (ALCAM/CD166), a member of the immunoglobulin superfamily with five extracellular immunoglobulin-like domains, facilitates heterophilic (ALCAM-CD6) and homophilic (ALCAM-ALCAM) cell-cell interactions. While expressed in a wide variety of tissues and cells, ALCAM is restricted to subsets of cells usually involved in dynamic growth and/or migration processes. ... More
Engagement of the common leukocyte antigen CD45 induces homotypic adhesion of activated human T cells.
AuthorsSpertini F, Wang AV, Chatila T, Geha RS
JournalJ Immunol
PubMed ID7913942
The transmembrane tyrosine phosphatase CD45 plays an important role in TCR/CD3-mediated signaling. We demonstrate in this study that ligand binding to the CD45 molecule induces homotypic cell adhesion of activated, but not resting, T lymphocytes. mAbs to CD45 (4B2 and 10G10) and to CD45RO (UCHL1), but not to CD45RA (IOL2), ... More
New fluorescent dyes for lymphocyte migration studies. Analysis by flow cytometry and fluorescence microscopy.
AuthorsWeston SA, Parish CR
JournalJ Immunol Methods
PubMed ID2212694
16 fluorochromes were examined for their ability to label viable lymphocytes in vitro and yield fluorescence detectable by fluorescence microscopy and flow cytometry. Of these fluorochromes, four intracellular dyes were found to be suitable for in vivo migration studies. They were H33342, the well known DNA-binding dye which excites and ... More
Distinct adhesive properties of granulocytes and monocytes to endothelial cells under static and stirred conditions.
AuthorsKuijpers TW, Hakkert BC, van Mourik JA, Roos D
JournalJ Immunol
PubMed ID1976697
Adherence of neutrophils and monocytes to endothelium is an important event in the sequence of leukocyte responses to inflammatory disease. Double-color flow microfluorimetry analysis was used to determine neutrophil and monocyte adherence to suspended endothelial cells under stirred conditions. The static adherence to endothelial cell monolayers was simultaneously determined. In ... More
Authorsvan de Wiel-van Kemenade E, Ligtenberg MJ, de Boer AJ, Buijs F, Vos HL, Melief CJ, Hilkens J, Figdor CG
JournalJ Immunol
PubMed ID7687622
Episialin (MUC1) is a mucin-like glycoprotein abundantly expressed on most carcinoma cells. As a result of its extended and rigid structure, it reduces intercellular adhesion. We investigated whether this antiadhesion function allows tumor cells expressing high levels of episialin to escape from immune recognition. To test this hypothesis, we transfected ... More
Membrane water and solute permeability determined quantitatively by self-quenching of an entrapped fluorophore.
AuthorsChen PY, Pearce D, Verkman AS
JournalBiochemistry
PubMed ID3179272
Quantitative determination of rapid water and solute transport and solute reflection coefficients by light-scattering methods is complicated by dependence of vesicle or cell light scattering on nonvolume factors including solution refractive index, cell motion, and membrane aggregation. To overcome these difficulties, a fluorescence technique has been developed to measure accurately ... More
ICAM-1 (CD54): a counter-receptor for Mac-1 (CD11b/CD18).
AuthorsDiamond MS, Staunton DE, de Fougerolles AR, Stacker SA, Garcia-Aguilar J, Hibbs ML, Springer TA
JournalJ Cell Biol
PubMed ID1980124
While the leukocyte integrin lymphocyte function-associated antigen (LFA)-1 has been demonstrated to bind intercellular adhesion molecule (ICAM)-1, results with the related Mac-1 molecule have been controversial. We have used multiple cell binding assays, purified Mac-1 and ICAM-1, and cell lines transfected with Mac-1 and ICAM-1 cDNAs to examine the interaction ... More
Intravital fluorescence microscopy of endothelial cells on vascular grafts.
AuthorsMechem C, Jarrell BE, Koolpe E, Williams SK
JournalJ Surg Res
PubMed ID2472511
The ability to evaluate the extent of initial endothelial cell coverage on a vascular graft subsequent to an endothelial cell seeding technique would be desirable to substantiate the durability of the endothelial cell lining. To this end, we have evaluated the use of intravital fluorescence microscopy to assess human endothelial ... More
Activation of LFA-1 through a Ca2(+)-dependent epitope stimulates lymphocyte adhesion.
Authorsvan Kooyk Y, Weder P, Hogervorst F, Verhoeven AJ, van Seventer G, te Velde AA, Borst J, Keizer GD, Figdor CG
JournalJ Cell Biol
PubMed ID1703161
The leukocyte function-associated molecule-1 (LFA-1) plays a key role in cell adhesion processes between cells of the immune system. We investigated the mechanism that may regulate LFA-1-ligand interactions, which result in cell-cell adhesion. To this end we employed an intriguing anti-LFA-1 alpha mAb (NKI-L16), capable of inducing rather than inhibiting ... More
Vitronectin receptor (alpha(v)beta3) mediates platelet adhesion to the luminal aspect of endothelial cells: implications for reperfusion in acute myocardial infarction.
AuthorsGawaz M, Neumann FJ, Dickfeld T, Reininger A, Adelsberger H, Gebhardt A, Schömig A
JournalCirculation
PubMed ID9323066
BACKGROUND: Platelet interaction with endothelium plays an important role in the pathophysiology of coronary microcirculation. We assessed the role of the vitronectin receptor (integrin alpha(v)beta3) in platelet/endothelium adhesion. METHODS AND RESULTS: We investigated the effect on platelet/endothelium adhesion of plasma obtained from patients with acute myocardial infarction during reperfusion (before ... More
Cognate interaction between human lymphocytes and eosinophils is mediated by beta 2-integrins and very late antigen-4.
AuthorsMengelers HJ, Maikoe T, Raaijmakers JA, Lammers JW, Koenderman L
JournalJ Lab Clin Med
PubMed ID7545203
Here the cognate interactions between human eosinophils and lymphocytes are studied. These interactions were measured in a double-colored FACS analysis by applying fluorescent red eosinophils (stained with hydroethidine, 40 mumol/L) and fluorescent green lymphocytes (stained with sulfidofluorescein diacetate, 100 mumol/L) in a ratio of 1:3. When normal eosinophils were mixed ... More
Stimulation of human basophils results in homotypic aggregation. A response independent of degranulation.
AuthorsKnol EF, Kuijpers TW, Mul FP, Roos D
JournalJ Immunol
PubMed ID7691958
For a better insight into the mechanisms determining the recruitment of human basophilic granulocytes from the circulation to sites of allergic reactions, we studied the homotypic aggregation of these cells. The aggregation was studied with > 95% pure basophil suspensions obtained from peripheral blood in a double-color flow cytometric analysis. ... More