Figure 4.

Copy number of foxP3, IL10 and TGFβ in CD3+ T cells from blood, lymph nodes, spleen, lung graft (left) and native lung (right) from normal dogs, chimeric recipients and chimeric recipients after lung transplantation at end of study. Copy number of foxP3, IL10 and TGFβ was measured by quantitative RT-PCR. Absolute copy numbers were calculated based on foxP3, IL10 or TGFβ standard curves, and samples were normalized to a second standard curve for the housekeeping gene G3PDH (37). A) Copy number of foxP3 was significantly increased in CD3+ T cells from the peripheral blood in recipients with mixed hematopoietic chimerism compared to normal dogs. Copy number of foxP3 from tissue-derived CD3+ T cells were not significantly different between normal dogs and chimeric recipients except for the right (native) lung in which it was noted to be increased in chimeric recipients. B) Copy number of IL10 was significantly increased in CD3+ T cells from the peripheral blood and left (graft) lung of recipients and increased but not significantly in the right (native) lung. There were significant decreases in IL10 copy number in CD3+ T cells from lymph node and spleen in the chimeric recipients. C) Copy number of TGFβ was significantly increased in CD3+ T cells from the peripheral blood of chimeric recipients (except after lung transplantation) and in the lungs (native and graft) after lung transplantation. There were significant decreases in TGFβ copy number in CD3+ T cells from lymph node and spleen in the chimeric recipients.