Figure 7: The defect in TFH induction in 6F/6F mice is T-cell intrinsic.
From: TCR ITAM multiplicity is required for the generation of follicular helper T-cells
(a–e) Bone marrow chimeras were generated by injecting a 1:1 mix of lineage-depleted 6Y/6Y and B6:CD45.1 cells or a 1:1 mix of lineage-depleted 6F/6F and B6.CD45.1 bone marrow cells into three lethally irradiated B6:CD45.1 mice. Six weeks later, mice were injected with sheep red blood cells (SRBC) and splenocytes were analysed 10 days after SRBC injection. (a) Dot plots show representative CXCR5 versus PD-1 staining of CD45.1-negative (6Y/6Y or 6F/6F) or CD45.1-positive (B6) CD4-SP splenocytes from 1:1 6Y/6Y:B6 or 1:1 6F/6F:B6 bone marrow chimeras. Bar graphs on right show results from all chimeric mice (3 6Y/6Y:B6 chimeras and 3 6F/6F:B6 chimeras). (b) Dot plots show representative Fas versus GL-7 staining of CD45.1-negative (6Y/6Y or 6F/6F) or CD45.1-positive (B6) B220+ splenocytes from 1:1 6Y/6Y:B6 or 1:1 6F/6F:B6 bone marrow chimeras. Bar graphs on right show results from all chimeric mice (three 6Y/6Y:B6 chimeras and three 6F/6F:B6 chimeras). (c) CD45.2/CD54.1 ratio of TFH and GC B-cells in 6Y/6Y:B6 and 6F/6F:B6 chimeras. (d) CD45.2/CD54.1 ratio of TFH (CD4+ CXCR5+ PD-1+), follicular Treg (CD4+ CXCR5+ PD-1+ Foxp3+) and Tregs (CD4+ Foxp3+) in 6Y/6Y:B6 and 6F/6F:B6 chimeras. (e) Percentage of TFH cells within all CD4+ Foxp3− splenocytes. (f) Age- and gender- matched 6Y/6Y and 6F/6F mice (14 of each) were infected with Listeria monocytogenes that expresses 2W peptide, and splenocytes were analysed by flow cytometry 10 days after infection. The number of 2W:I-Ab-specific splenocytes was determined by tetramer enrichment. Th1 (CD4+IFNγ+), TFH (CD4+ CXCR5+PD-1+), GC-TFH (CD4+CXCR5hi PD-1hi). For bar graphs, data were analysed by unpaired t-test (two tailed) and are represented as mean±s.d. *P<0.05; ***P<0.001.
