Figure 4

Depletion of CD4⁺ and CD8⁺ T-cells and altered cytokine/chemokines expression in peripheral blood of Tat-Tg mice.

(A) Percentage and (B) absolute number of lymphocytes in peripheral blood of 8 weeks old wild type and Tat-Tg mice. Values (mean ± SE, n = 6) are shown. (C) Whole blood cells (1 × 10⁶) of 8 weeks old wild type and Tat-Tg mice were stained with anti-CD3-FITC and anti-B220-PE antibodies and analysed by flow cytometry. Percentages of CD3⁺ T-lymphocytes (upper dot plots) and B220⁺ B-lymphocytes (bottom dot plots) of wild type and Tat-Tg mice are shown. Each plot represents 10,000 events of a representative experiment. (D) Absolute numbers of CD3⁺ T-lymphocytes and B220⁺ B-lymphocytes of wild type and Tat-Tg mice. Values (mean ± SE, n = 6) are shown. The asterisk indicates a statistically significant difference according to the Student’s t test (p < 0.01). (E) Whole blood cells (1 × 10⁶) of 8 weeks old wild type and Tat-Tg mice were stained with anti-CD3-APC, anti-CD45-V500, anti-CD14-V450, anti-CD4-PE and anti-CD8-FITC and analysed by flow cytometry. Percentages of CD3⁺ CD4⁻CD8⁻, CD3⁺CD4⁺CD8⁺, CD3⁺CD4⁺CD8⁻ and CD3⁺CD4⁻CD8⁺ of peripheral blood populations of wild type and Tat-Tg mice are shown. Each plot represents 20,000 events of a representative experiment. (F) Absolute number of CD3⁺ CD4⁺ CD8⁻ and CD3⁺ CD4⁻CD8⁺ cells. Values (mean ± SE, n = 6) are shown. The asterisk indicates a statistically significant difference according Student’s t test (p < 0.01). (G) Whole blood cells (1 × 10⁶) of 8 weeks old wild type and Tat-Tg mice were stained with anti-CD3-APC-Cy7, anti-CD4-PE, anti-CD8-FITC, Annexin V-APC and 7-AAD and analysed by flow cytometry. Percentage of apoptotic CD3⁺ CD4⁺ and CD3⁺ CD8⁺ cells in wild type and Tat-Tg mice are shown. (H) Absolute number of apoptotic CD3⁺ CD4⁺ and CD3⁺ CD8⁺ cells in wild type and Tat-Tg mice are shown. Values (mean ± SE, n = 6) are shown. (I) Serum samples were obtained from retro-orbital blood of 8-weeks old wild type and Tat-Tg mice and analysed for cytokines and chemokines expression, using the Mouse Cytokine Array Panel A (R&D System). Values (mean ± SE, n = 3) are shown. The asterisk indicates a statistically significant difference according Student’s t test (p < 0.01).