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Pathogenesis and Immunity

Differential Expression of Interleukin-2 and Gamma Interferon in Human Immunodeficiency Virus Disease

Scott F. Sieg, Douglas A. Bazdar, Clifford V. Harding, Michael M. Lederman
Scott F. Sieg
Department of Medicine, Division of Infectious Diseases, and
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Douglas A. Bazdar
Department of Medicine, Division of Infectious Diseases, and
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Clifford V. Harding
Department of Pathology, Center for AIDS Research, Case Western Reserve University, Cleveland, Ohio 44106
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Michael M. Lederman
Department of Medicine, Division of Infectious Diseases, and
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DOI: 10.1128/JVI.75.20.9983-9985.2001
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  • Fig. 1.
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    Fig. 1.

    Differential production of IL-2 and IFN-γ in CD69+ cells. Whole blood was incubated with SEB (Sigma) (5 μg/ml) for 2 h before Golgi-stop reagent (BD Pharmingen) containing brefeldin A was added. Cells were incubated for an additional 3 h, at which time the cells were treated with Facs lyse solution (Becton Dickinson) and frozen at −86°C overnight. Frozen cells were thawed and treated with Facs permeabilizing solution (Becton Dickinson) and then stained with anti-CD69 phycoerythrin-conjugated antibody, anti-CD4 peridinin chlorophyll protein-conjugated antibody, and either anti-IL-2 or anti-IFN-γ fluorescein isothiocyanate-conjugated antibody (Becton Dickinson). Cells stained with isotype control antibodies were used to establish quadrants. Unstimulated cells in this case were exposed to anti-CD28 antibody (1 μg/ml) to control for additional experiments in which cells were stimulated with SEB plus anti-CD28 antibody (data not shown). The numbers shown indicate the percentages of CD69+ cells that were IL-2 positive following SEB stimulation.

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    Fig. 2.

    Reduced proportions of IFN-γ-producing cells make IL-2 in CD4+ T cells from HIV-infected patients. Whole blood was stimulated and processed as described in the legend to Fig. 1 except that cells were stained with anti-CD4 peridinin chlorophyll protein-conjugated antibody, anti-IFN-γ phycoerythrin-conjugated antibody, and anti-IL-2 fluorescein isothiocyanate-conjugated antibody. The numbers shown indicate the percentages of IFN-γ-producing cells that were IL-2 positive following SEB stimulation.

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    Fig. 3.

    Reduced IL-2 production in CMV-stimulated cells. Whole blood was stimulated with CMV antigen (3 μg/ml) with the addition of anti-CD28 and anti-CD49d antibodies (BD Pharmingen) at 3 μg/ml and analyzed by three-color flow cytometry. Unstimulated cells were exposed to costimulatory antibodies alone. Cells were gated on CD4+cells, and histograms were generated based on the stains shown.

Tables

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  • Table 1.

    Expression of IL-2 and IFN-γ following SEB stimulationa

    Patient or controlHIV RNA in plasma (no. of copies/ml)No. of CD4 cells/μlCD69+ % IL-2+CD69+ % IFN-γ+IFN-γ+ % IL-2+
    Patients
     536<40048615.125.5ND
     24815,00020211.532.112.3
     13821,00030911.826.512.4
     12125,0004606.436.15.9
     26132,0006809.0ND21
     11365,00025021.752.65.7
     09197,000190922
     107520,00011211.712.324
     Mean12.0∗ 29.613.6†
     Median28,500275.911.626.512.35
    Controls
     116.717.134.4
     211.224.832
     319.425.443.2
     49.74839
     52229ND
     618.325.529
     725NDND
     821ND20.0
     Mean17.928.333.0
     Median18.325.533.2
    • ↵a P values from two-tailedt tests of comparisons between the average responses of patients and healthy donors are as follows: ∗, P < 0.03 ; †, P < 0.001 . ND, not done.

  • Table 2.

    Expression of IL-2 and IFN-γ following CMV stimulation

    Patient or controlHIV RNA in plasma (no. of copies/ml)No. of CD4 cells/μlCD69+ % IL-2+CD69+ % IFN-γ+IFN-γ+ % IL-2+
    Patients
     14123,00059820.590.224
     150135,00034410859
     26691,0007845708
     265196,0006313.55322
     16493,00010411.045NDa
     19570,000464NDND33.6
     Mean101,00039312.068.619.3
    Controls
     1576241
     2245326
     319.450ND
     4NDND62
     Mean33.55543
    • ↵a ND, not done.

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Differential Expression of Interleukin-2 and Gamma Interferon in Human Immunodeficiency Virus Disease
Scott F. Sieg, Douglas A. Bazdar, Clifford V. Harding, Michael M. Lederman
Journal of Virology Oct 2001, 75 (20) 9983-9985; DOI: 10.1128/JVI.75.20.9983-9985.2001

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Differential Expression of Interleukin-2 and Gamma Interferon in Human Immunodeficiency Virus Disease
Scott F. Sieg, Douglas A. Bazdar, Clifford V. Harding, Michael M. Lederman
Journal of Virology Oct 2001, 75 (20) 9983-9985; DOI: 10.1128/JVI.75.20.9983-9985.2001
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KEYWORDS

CD4-Positive T-Lymphocytes
HIV Infections
Interferon-gamma
Interleukin-2

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