Figure 3
Figure 3. Db-SAP tetramers activate and eliminate antigen-specific T cells in vitro. (A) Incubation of a 50:50 mixture of female Vα2+ (P14) and T3.70+ (H-Y) CD8+ T cells with gpC9M-SAP or hyC2A-SAP results in the selective loss of cognate T cells. Tetramer mixtures are indicated by the legends above the dotplots. Following an initial 2-hour incubation with tetramer, cells were washed repeatedly and then cultured for an additional 46 hours prior to harvest and staining. For each group, 104 events were analyzed; the plots show only live (7-AAD−) CD8α-gated cells. Numbers indicate the cell number in the region. (B) Loss of T cells with Db-SAP tetramer treatment is not the result of inadequate stimulation through the TCR. Exposure to gpC9M or gpC9M-SAP causes equivalent changes in CD62L and CD69 expression, and identical increases in ΔΨm (measured as increased DiIC\batchmode \documentclass[fleqn,10pt,legalpaper]{article} \usepackage{amssymb} \usepackage{amsfonts} \usepackage{amsmath} \pagestyle{empty} \begin{document} \(_{1}^{<CROSS-REF\ REFID="B5"\ TYPE="BIB">5</CROSS-REF>}\) \end{document} fluorescence), characteristic of antigen-activated T cells. P14 T cells were analyzed after 12 hours of culture; equivalent activation was also found at 8- and 16-hour time points (not shown). Histograms are gated on 7-AAD− cells, which are present in equal percentages in tetramer-treated groups. Identical results were obtained in 4 (A) or 2 (B) independent experiments.

Db-SAP tetramers activate and eliminate antigen-specific T cells in vitro. (A) Incubation of a 50:50 mixture of female Vα2+ (P14) and T3.70+ (H-Y) CD8+ T cells with gpC9M-SAP or hyC2A-SAP results in the selective loss of cognate T cells. Tetramer mixtures are indicated by the legends above the dotplots. Following an initial 2-hour incubation with tetramer, cells were washed repeatedly and then cultured for an additional 46 hours prior to harvest and staining. For each group, 104 events were analyzed; the plots show only live (7-AAD−) CD8α-gated cells. Numbers indicate the cell number in the region. (B) Loss of T cells with Db-SAP tetramer treatment is not the result of inadequate stimulation through the TCR. Exposure to gpC9M or gpC9M-SAP causes equivalent changes in CD62L and CD69 expression, and identical increases in ΔΨm (measured as increased DiIC

\(_{1}^{<CROSS-REF\ REFID="B5"\ TYPE="BIB">5</CROSS-REF>}\)
fluorescence), characteristic of antigen-activated T cells. P14 T cells were analyzed after 12 hours of culture; equivalent activation was also found at 8- and 16-hour time points (not shown). Histograms are gated on 7-AAD− cells, which are present in equal percentages in tetramer-treated groups. Identical results were obtained in 4 (A) or 2 (B) independent experiments.

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