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Blood, Vol. 110, Issue 4, 1207-1214, August 15, 2007
HIV modulates the expression of ligands important in triggering natural killer cell cytotoxic responses on infected primary T-cell blasts
Blood Ward et al.
110: 1207
Supplemental materials for Ward et al, Vol. 110, Issue 4, 1207-1214
Files in this Data Supplement:
- Figure S1. Differing abilities of two clones of anti-HIV p24 mAbs to detect antigen (JPG, 68 KB)
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Primary CD4+ PHA blasts were infected with 104 TCID50 HIV-1SF162 or left uninfected in culture. Following HIV infection, aliquots from the same culture were stained for CD4, perm/fixed, and intracellularly stained for HIV p24 antigen with either the human 71-31 clone of anti-p24 monoclonal antibody (mAb) followed by secondary antibody (A), or the mouse KC57 clone of anti-p24 mAb (B). In a subsequent experiment, primary phytohemagglutinin (PHA) blasts from a different donor were infected with 103 TCID50 HIV1SF162 and stained as in Figure A for CD4 and the 71-31 anti-p24 mAb (C).
- Figure S2. Purity of CD4−/p24 Ag+ infected populations used for 51Cr release assays (JPG, 56.5 KB)
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CD4+ PHA blasts were infected with HIV-1SF128A and depleted of CD4+ cells by negative selection with anti-CD4 coated beads. The isolated and uninfected cells that were not CD4-bead isolated were surface stained for CD4, perm/fixed, and intracellularly stained for HIV p24 using a human anti-p24 mAb (clone 71-31) and a goat antihuman secondary. All other flow cytometry plots presented in the written manuscript were of bulk HIV-1 infected cultures and not purified for HIV-infected cells as done here.
- Figure S3. Relative percentages of NKG2D/NKG2A double positive CD56+CD3− NK cells of healthy donors (JPG, 44.1 KB)
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Peripheral blood mononuclear cells (PBMCs) from healthy donors that were used to generate the data presented in Figure 5 were stained for CD56, CD3, NKG2D, and NKG2A. Dot plots shown are of 104 gated CD56+ CD3− NK cells. The donor of the cells shown in Figure S3A is the same as the donor of the cells used in the cytotoxic assay presented in Figure 5A; the cell donor is the same in Figures S3B and 5B, and in Figures S3C and 5C,D.
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