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Blood, Vol. 114, Issue 3, 564-571, July 16, 2009
Enhanced development of CD4+   T cells in the absence of Itk results in elevated IgE production
Blood Qi et al.
114: 564
Supplemental materials for: Qi et al
Files in this Data Supplement:
- Figure S1. Enhanced development of γδ T cells in the absence of Itk is bone marrow intrinsic (JPG, 191 KB)
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Data from the experiments shown in Fig. 3. (A) Percentage of CD3+γδTCR+ cells of donor WT and Itk−∕− cells. *p<0.001, n=6, between WT and Itk donors in both thymus and spleen. (B) Percentage of CD4+ population of donor γδ T cells. *p<0.001, n=3, between WT and Itk−∕− donors in thymus (bottom panel). (C) Percentage of CD3+αβTcR+ or CD3+γδTcR+ cells of donor WT and Itk−∕− origin. *p<0.001, n=3, between WT and Itk−∕− CD3+αβTcR+ cells in the thymus; p=0.018, n=3, between WT and Itk−∕− CD3+αβTcR+ cells in the spleen; p=0.033, n=3, between WT and Itk−∕− CD3+γδTcR+ cells in the thymus, p=0.01, n=3, between WT and Itk−∕− CD3+γδTcR+ cells in the spleen.
- Figure S2. γδ TcR expression of γδ T cells from WT and Itk−∕− mice (JPG, 426 KB)
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Thymocytes and splenocytes from WT and Itk−∕− mice were gated on γδ T cells and analyzed for the expression of (A) Vγ2 vs. γδ TcR (left), Vγ2 vs. CD4 (right); (B) NK1.1 vs. Vδ6.2/3 (left), CD4 vs. Vδ6.2/3 (right); or (C) Vγ1.1 vs. CD4. Data are representative of two independent experiments.
- Figure S3. Itk null CD4+ γδ T cells predominantly express Vγ1.1/δ6.2/3 TcR but develop independent of the γδ TcR they express (JPG, 68.8 KB)
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Data from the experiments shown in Fig. 5. (A) Percentage of Vγ1.1/Vδ6.2/3+CD4+γδ T cells in the thymus and spleens of WT and Itk−∕− mice. *p<0.01, n=6, between WT and Itk−∕− Vγ1.1/Vδ6.2/3+CD4+γδ T cells in the spleen or thymus. (B) Percentage of Vγ2+ transgenic+CD4+ T cells in the thymus and spleens of KN6 transgenic mice, or KN6 transgenic mice lacking Itk. *p=0.02, n=3, between WT and Itk−∕− KN6 transgenic thymus, p=0.03, n=3, between WT and Itk−∕− KN6 transgenic spleen.
- Figure S4. Itk−∕− γδ T cells carry increased IL-4 mRNA (JPG, 407 KB)
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(A) CD4+ or CD4− CD3+γδTCR+ cells were sorted from the thymus of WT and Itk−∕− mice and mRNA for T-bet, eomesodermin, GATA-3, IFNγ and IL-4 were analyzed by quantitative real-time PCR. (B) CD4+ or CD4− CD3+γδTCR+ cells from the spleens of WT and Itk−∕− mice were sorted and mRNA for T-bet, eomesodermin, GATA-3, IFNγ, and IL-4 were analyzed by quantitative real-time PCR. Data were analyzed using the comparative threshold cycle δδCT method, normalized to the expression of GAPDH and the relative gene expression levels were determined as fold increase compared to WT CD4− γδ T cells, which was set as 1. Data are representative of two independent experiments.
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