Extended Data Fig. 3. The spatial organization of the GC is disrupted in aged C57BL/6 mice.
(a) Representative confocal images of GCs at 20x from adult and aged C57BL/6 mice 12 days after immunization with NP-OVA in alum. Scale bars are 100 µm. LN sections were stained for IgD (green), CD35 (white), Ki67 (blue) and CD3 (magenta). Enumeration of GC number (b) and area (c) per LN was performed by examining 6–10 sections throughout each LN and identifying GCs as CD35+Ki67+IgD− structures. (d) Quantification of the CD35+ FDC network light zone area of the GC. (e) Quantification of the Ki67+CD35− dark zone area of the GC. (f) Quantification of the number of CD3+ T cells identified within the total Ki67+CD35+IgD− GC area. Quantification of the proportion of T cells positioned in the CD35+ FDC light zone area (g) and Ki67+CD35− dark zone (h) of the GC (n = 15). (i) Representative confocal images of GCs at 40x from unimmunized naive adult and aged C57BL/6 mice and (j) quantification of the CD35+ FDC network in the primary B cell follicle (n = 11). For (b-h, j), quantification of the GC compartments and T cell positioning was performed using an automated Cell Profiler pipeline. The data is representative of two independent experiments where each symbol on the graph represents a mouse and the bar height represents the median. The p-values were generated by performing an unpaired, two-tailed Mann Whitney U test.