Article Figures & Data
Figures
- FIGURE 1.
Specificity of the anti-gp49 mAb H1.1. A, The indicated cells were incubated with H1.1 FITC mAb or B23.1 mAb, followed by goat F(ab′)2 anti-mouse IgG FITC. Solid lines represent specific staining, and dotted lines indicate staining with no Ab (H1.1 staining) or secondary alone (B23.1). The indicated cell lines did not react with FITC-conjugated isotype control Ab for H1.1, hamster IgG (data not shown). All of the FACS profiles depict the log of the mean fluorescence intensity. B, KY-2 cells or LAK cells derived from C57BL/6 mice were stained by mAb H1.1 FITC.
- FIGURE 2.
Cellular distribution of the gp49 receptor in C57BL/6 mice. Single cell suspensions from the indicated tissue were prepared from C57BL/6 mice and stained with the indicated mAbs. Depicted is the flow cytometry data from one representative mouse. The percentage of cells in each quadrant is indicated.
- FIGURE 3.
Expression of the gp49 receptor on in vivo activated NK cells. A, C57BL/6 mice were infected with 7.5 × 104 PFU of MCMV. Three days postinfection, cells from the indicated tissue were harvested and subjected to flow cytometry with the indicated mAbs. B, NK1.1+ spleen- and liver-derived lymphocytes from MCMV-infected mice were assessed at various days postinfection for expression of gp49 (top panel) or CD69 (bottom panel). Depicted is the average of the mean fluorescence intensity from four mice at each time point of gp49 or CD69 staining on NK1.1+ cells.
- FIGURE 4.
Inducible expression of gp49 and CD69 in IL-12−/− or IFN-γ−/− mice. Mice with the indicated targeted mutation or C57BL/6 control mice were infected with 5 × 104 PFU of MCMV. Three days postinfection, splenocytes were harvested and subjected to flow cytometry with the indicated mAbs. NK cells from uninfected IL-12−/− or IFN-γ−/− mice do not express gp49 (data not shown).
- FIGURE 5.
Expression of gp49 isoforms by RT-PCR analysis. cDNA from the indicated cell types were amplified with primers specific for gp49A, gp49B, or HPRT, and analyzed by 1.2% agarose gel electrophoresis and ethidium bromide staining. The predicted molecular sizes of the products from the endogenous cDNA are: HPRT (176 bp), gp49A (577 bp), and gp49B (576 bp). Positive control cDNA templates used were a truncated gp49A, resulting in a 100-bp smaller PCR product, and full-length gp49B.
- FIGURE 6.
mAb-mediated inhibition of cytokine release by KY-2 cells. KY-2 cells were incubated for 5 h with the indicated biotinylated mAbs bound to plate-immobilized avidin. Cell-free supernatants were assayed for GM-CSF levels by ELISA and quantitated by extrapolation to a standard curve with known GM-CSF concentrations. Background GM-CSF release by KY-2 cells incubated with plate-immobilized avidin alone was 67 pg/ml. Depicted is one representative experiment. This assay was performed three times, and similar levels of inhibition were observed in each experiment.
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