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  1. Article ; Online: Heavy metal contaminants can eliminate quantum dot fluorescence.

    Zarkowsky, David / Lamoreaux, Laurie / Chattopadhyay, Pratip / Koup, Richard A / Perfetto, Stephen P / Roederer, Mario

    Cytometry. Part A : the journal of the International Society for Analytical Cytology

    2010  Volume 79, Issue 1, Page(s) 84–89

    Abstract: Quantum dots (QD) are fluorescent nanocrystals that are highly useful in imaging and flow cytometric analyses. During routine use of monoclonal antibody conjugates of QD, we have occasionally seen partial or total loss of fluorescence when using certain ... ...

    Abstract Quantum dots (QD) are fluorescent nanocrystals that are highly useful in imaging and flow cytometric analyses. During routine use of monoclonal antibody conjugates of QD, we have occasionally seen partial or total loss of fluorescence when using certain lots of fixative solutions. We hypothesized that a low level contamination with heavy metal cations was responsible, since low level metal contaminants are not uncommon in formalin solutions. By titrating known concentrations of heavy metal cations into staining solutions, we found that millimolar concentrations of ferrous and zinc ions, and as low as 50 nanomolar cupric ions, completely eliminated QD fluorescence. By mass spectroscopic quantification of metals in commercial fixative solutions previously shown to perform poorly or well with regard to QD fluorescence, we confirmed that the presence of copper in solution was correlated with poor performance. Notably, prior addition of EDTA to chelate the divalent cations in these solutions prevented the inhibition of QD fluorescence. Finally, the copper-induced loss of QD fluorescence is irreversible: cells labeled with QD are highly fluorescent and can be rendered nonfluorescent by the addition of cupric sulfate, even after washing extensively. Indeed, these cells can then be successfully stained with other QD reagents, providing a method for immunofluorescence restaining of cells without contaminating fluorescence from the first stain.
    MeSH term(s) Copper/analysis ; Fixatives/chemistry ; Flow Cytometry/methods ; Fluorescence ; Formaldehyde ; Humans ; Indicators and Reagents ; Ions ; Iron/analysis ; Leukocytes, Mononuclear/chemistry ; Quantum Dots ; Zinc/analysis
    Chemical Substances Fixatives ; Indicators and Reagents ; Ions ; Formaldehyde (1HG84L3525) ; Copper (789U1901C5) ; Iron (E1UOL152H7) ; Zinc (J41CSQ7QDS)
    Language English
    Publishing date 2010-11-11
    Publishing country United States
    Document type Journal Article ; Research Support, N.I.H., Intramural
    ZDB-ID 2099868-5
    ISSN 1552-4930 ; 0196-4763 ; 1552-4922
    ISSN (online) 1552-4930
    ISSN 0196-4763 ; 1552-4922
    DOI 10.1002/cyto.a.20986
    Database MEDical Literature Analysis and Retrieval System OnLINE

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    In stock of ZB MED Cologne/Königswinter

    Zs.A 1688: Show issues Location:
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    ab Jg. 2022: Lesesaal (EG)
    Zs.MG 68: Show issues

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  2. Article ; Online: Replication-defective adenovirus vectors with multiple deletions do not induce measurable vector-specific T cells in human trials.

    Koup, Richard A / Lamoreaux, Laurie / Zarkowsky, David / Bailer, Robert T / King, C Richter / Gall, Jason G D / Brough, Douglas E / Graham, Barney S / Roederer, Mario

    Journal of virology

    2009  Volume 83, Issue 12, Page(s) 6318–6322

    Abstract: The magnitude and character of adenovirus serotype 5 (Ad5)-specific T cells were determined in volunteers with and without preexisting neutralizing antibodies (NAs) to Ad5 who received replication-defective Ad5 (rAd5)-based human immunodeficiency virus ... ...

    Abstract The magnitude and character of adenovirus serotype 5 (Ad5)-specific T cells were determined in volunteers with and without preexisting neutralizing antibodies (NAs) to Ad5 who received replication-defective Ad5 (rAd5)-based human immunodeficiency virus vaccines. There was no correlation between T-cell responses and NAs to Ad5. There was no increase in magnitude or activation state of Ad5-specific CD4(+) T cells at time points where antibodies to Ad5 and T-cell responses to the recombinant gene products could be measured. These data indicate that rAd5-based vaccines containing deletions in the E1, E3, and E4 regions do not induce appreciable expansion of vector-specific CD4(+) T cells.
    MeSH term(s) AIDS Vaccines/immunology ; Adenovirus Infections, Human/immunology ; Adenovirus Infections, Human/virology ; Adenoviruses, Human/genetics ; Adenoviruses, Human/immunology ; Antibodies, Viral/blood ; Antibodies, Viral/immunology ; CD4-Positive T-Lymphocytes/immunology ; CD8-Positive T-Lymphocytes/immunology ; Defective Viruses/genetics ; Gene Deletion ; Genes, Viral ; Genetic Vectors ; HIV Infections/immunology ; HIV Infections/prevention & control ; HIV-1/immunology ; Humans ; Lymphocyte Activation/immunology ; Neutralization Tests ; env Gene Products, Human Immunodeficiency Virus/immunology
    Chemical Substances AIDS Vaccines ; Antibodies, Viral ; env Gene Products, Human Immunodeficiency Virus
    Language English
    Publishing date 2009-04-01
    Publishing country United States
    Document type Journal Article ; Research Support, N.I.H., Intramural ; Research Support, Non-U.S. Gov't
    ZDB-ID 80174-4
    ISSN 1098-5514 ; 0022-538X
    ISSN (online) 1098-5514
    ISSN 0022-538X
    DOI 10.1128/JVI.00384-09
    Database MEDical Literature Analysis and Retrieval System OnLINE

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    In stock of ZB MED Cologne/Königswinter

    Zs.A 609: Show issues Location:
    Je nach Verfügbarkeit (siehe Angabe bei Bestand)
    bis Jg. 1994: Bestellungen von Artikeln über das Online-Bestellformular
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  3. Article ; Online: Phenotypic and functional profile of HIV-inhibitory CD8 T cells elicited by natural infection and heterologous prime/boost vaccination.

    Freel, Stephanie A / Lamoreaux, Laurie / Chattopadhyay, Pratip K / Saunders, Kevin / Zarkowsky, David / Overman, R Glenn / Ochsenbauer, Christina / Edmonds, Tara G / Kappes, John C / Cunningham, Coleen K / Denny, Thomas N / Weinhold, Kent J / Ferrari, Guido / Haynes, Barton F / Koup, Richard A / Graham, Barney S / Roederer, Mario / Tomaras, Georgia D

    Journal of virology

    2010  Volume 84, Issue 10, Page(s) 4998–5006

    Abstract: Control of HIV-1 replication following nonsterilizing HIV-1 vaccination could be achieved by vaccine-elicited CD8(+) T-cell-mediated antiviral activity. To date, neither the functional nor the phenotypic profiles of CD8(+) T cells capable of this ... ...

    Abstract Control of HIV-1 replication following nonsterilizing HIV-1 vaccination could be achieved by vaccine-elicited CD8(+) T-cell-mediated antiviral activity. To date, neither the functional nor the phenotypic profiles of CD8(+) T cells capable of this activity are clearly understood; consequently, little is known regarding the ability of vaccine strategies to elicit them. We used multiparameter flow cytometry and viable cell sorts from phenotypically defined CD8(+) T-cell subsets in combination with a highly standardized virus inhibition assay to evaluate CD8(+) T-cell-mediated inhibition of viral replication. Here we show that vaccination against HIV-1 Env and Gag-Pol by DNA priming followed by recombinant adenovirus type 5 (rAd5) boosting elicited CD8(+) T-cell-mediated antiviral activity against several viruses with either lab-adapted or transmitted virus envelopes. As it did for chronically infected virus controllers, this activity correlated with HIV-1-specific CD107a or macrophage inflammatory protein 1beta (MIP-1beta) expression from HIV-1-specific T cells. Moreover, for vaccinees or virus controllers, purified memory CD8(+) T cells from a wide range of differentiation stages were capable of significantly inhibiting virus replication. Our data define attributes of an antiviral CD8(+) T-cell response that may be optimized in the search for an efficacious HIV-1 vaccine.
    MeSH term(s) AIDS Vaccines/immunology ; Adenoviridae/genetics ; CD8-Positive T-Lymphocytes/immunology ; Flow Cytometry ; HIV Infections/immunology ; HIV-1/immunology ; Humans ; Immunization, Secondary ; T-Lymphocyte Subsets/immunology ; Transduction, Genetic ; Vaccination/methods ; Vaccines, DNA/immunology
    Chemical Substances AIDS Vaccines ; Vaccines, DNA
    Language English
    Publishing date 2010-03-03
    Publishing country United States
    Document type Journal Article ; Research Support, N.I.H., Extramural ; Research Support, N.I.H., Intramural
    ZDB-ID 80174-4
    ISSN 1098-5514 ; 0022-538X
    ISSN (online) 1098-5514
    ISSN 0022-538X
    DOI 10.1128/JVI.00138-10
    Database MEDical Literature Analysis and Retrieval System OnLINE

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    In stock of ZB MED Cologne/Königswinter

    Zs.A 609: Show issues Location:
    Je nach Verfügbarkeit (siehe Angabe bei Bestand)
    bis Jg. 1994: Bestellungen von Artikeln über das Online-Bestellformular
    Jg. 1995 - 2021: Lesesall (1.OG)
    ab Jg. 2022: Lesesaal (EG)

    Order via subito

    This service is chargeable due to the Delivery terms set by subito. Orders including an article and supplementary material will be classified as separate orders. In these cases, fees will be demanded for each order.

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