Fig. 2.
Fig. 2. Development of CD4ζ/γ-expressing thymocytes and T cells is rescued in MHCII− mice. / Bone marrow cells isolated from MHCII− mice were exposed to retroviral supernatant encoding CD4ζ, CD4, or CD4del (CD4−), and then infused into sublethally irradiated wild-type C57BL/6 (A) or MHCII− C57BL/6 (B) mice. Six weeks after transplantation, peripheral blood was isolated and analyzed by 2-color flow cytometry using PE-conjugated antihuman CD4 and FITC-conjugated anti-B220 or anti-CD3. The percentage of lineage positive cells expressing human CD4 is shown in the top left hand corner of each dot plot. Control cells stained with antihuman CD4-PE or isotype controls yielded indistinguishable results (data not shown). Results are representative of at least 8 additional mice. (C) Thymocytes were isolated from MHCII− mice in panel B, which had received CD4ζ- or CD4del (CD4−)–transduced bone marrow, and were analyzed by 4-color flow cytometry as described in the legend to Figure 1. The percentage of human CD4-expressing cells (solid line) in each TN and TP subset is indicated in each histogram. Shaded histograms represent cells from control animals that received transplants of unmodified bone marrow in each case. ND indicates insufficient cells to analyze.

Development of CD4ζ/γ-expressing thymocytes and T cells is rescued in MHCII− mice.

Bone marrow cells isolated from MHCII− mice were exposed to retroviral supernatant encoding CD4ζ, CD4, or CD4del (CD4−), and then infused into sublethally irradiated wild-type C57BL/6 (A) or MHCII− C57BL/6 (B) mice. Six weeks after transplantation, peripheral blood was isolated and analyzed by 2-color flow cytometry using PE-conjugated antihuman CD4 and FITC-conjugated anti-B220 or anti-CD3. The percentage of lineage positive cells expressing human CD4 is shown in the top left hand corner of each dot plot. Control cells stained with antihuman CD4-PE or isotype controls yielded indistinguishable results (data not shown). Results are representative of at least 8 additional mice. (C) Thymocytes were isolated from MHCII− mice in panel B, which had received CD4ζ- or CD4del (CD4−)–transduced bone marrow, and were analyzed by 4-color flow cytometry as described in the legend to Figure 1. The percentage of human CD4-expressing cells (solid line) in each TN and TP subset is indicated in each histogram. Shaded histograms represent cells from control animals that received transplants of unmodified bone marrow in each case. ND indicates insufficient cells to analyze.

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