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. 2016 May 10;7:11514. doi: 10.1038/ncomms11514

Figure 4. Hair follicle clusters account for the increase in skin CD4 T cells.

Figure 4

(a) Hair follicle cluster frequency; n=8–15 and (b) number of CD4 T cells per cluster; n=107–167 clusters from seven to eight mice. (c) Number of inter-follicular dermal CD4 T cells; n=10–12. (d,e) HSV memory skin detecting (d) CD4 (orange) and CD11c (green) and (e) CD4 (orange) and CD8 (green). Scale bar, 100 μm. (f) Number of peri-follicular clusters incorporating pertussis toxin treated in vitro activated CD4 T cells, 1 week after intradermal injection into the skin of naive mice; n=6–9. (g) Chemokine mRNA transcripts in HSV memory skin relative to control skin (days 30–40); n=8. (h,i) HSV memory mice treated with an anti-CCL5 antibody before analysis of (h) gDT-II cells and (i) total CD4 T cells; n=7 and 20–24, respectively. (j) Hair follicle cluster frequency and number of CD4 T cells per cluster on the infected side of anti-CCL5-treated HSV memory mice; n=6 mice or 61–82 clusters pooled from six mice, respectively. (k) Enumeration of CD4+ T cells following intradermal injection of recombinant mouse CCL5. Symbols in (a,c,f,h,i; left graph of j and k) represent a single mouse. In (b and the right graph of (j) symbols represent the number of CD4+ T cells in a single peri-follicular cluster pooled from seven to eight and six mice respectively, from two experiments. NS, not significant; * P<0.05, ** P<0.01, *** P<0.001, **** P<0.0001 (two-tailed unpaired t-test (right of i, left of j and k) or two-tailed Mann–Whitney test (a–c,f,h; right of i; and left of j,k). Images (d,e) representative of four experiments with two to four mice. Data pooled from two to four (a,c,f), two (h,j,k) or seven (i) experiments with two to five mice. Mean (a–c,f,h–k), mean and s.e.m. (g). Iso; isotype control antibody; PTX, Pertussis toxin, rm; recombinant mouse.