Fig. 9
Aging increases the frequency of GM-CSF+ cells among CD4+ T lymphocytes in draining lymph nodes from AO rats immunized for EAE. (Panel a) Overlaid Flow cytometry histograms show GM-CSF expression in CD4+ T cells isolated from draining lymph nodes of (left) young and (right) aged rats on the 7th d.p.i. using MACS (described in detail in Methods). In the overlaid flow cytometry histograms left histograms (thin grey line) represent nonspecific binding of secondary antibody. Flow cytometry dot plots show IFN-γ vs IL-17 staining of GM-CSF+ T cells retrieved from draining lymph nodes of (left) young and (right) aged rats on the 7th d.p.i. Numbers in the flow cytometry profiles represent the percentage of cells in the indicated region. (Panel b) Left bar graphs show the fold change in expression of mRNAs for (upper) GM-CSF and (lower) IL-3 in CD4+ draining lymph node cells isolated on the 7th d.p.i. using MACS (described in detail in the section Methods) from aged relative to young rats. Right bar graphs show the fold change in expression of mRNAs for (upper) GM-CSF and (lower) IL-3 in draining lymph node cells cultivated for 72 h in RMPI medium from aged rats and MBP-stimulated cells from young and aged rats relative to those from young rats cultivated in RMPI medium, as determined by RT-qPCR. (c) Bar graph shows fold change in mRNA expression for IL-7 in draining lymph node tissue on the 7th d.p.i. from aged relative to young rats as determined by RT-qPCR. Data are normalized to β-actin (ACTB). All results are presented as means ± SEM (n = 9/group). The data, except for GM-CSF, are representative of one of two experiments with similar results. **p < 0.01; ***p < 0.001