Fig 5. Suppressive Tregs and Tfr in normal subjects and low-grade lymphoma.

(A) Biaxial flow cytometry plot showing expression of CD4 and FOXP3 on cells gated for CD4⁺CD45RA^-. The highest 5% of cells from 11 concatenated NS were employed for the FOXP3^hi gate. This representative example from a patient with lymphoma shows Treg 5.9% of CD4⁺CD45RA^- cells. (B) Percentage of Treg within total CD4⁺ T-cells in normal subjects (n = 11), MZL (N = 4) and BNHL (n = 9). Horizontal lines represent the median and bars represent inter-quartile range. No significant differences were found between groups (Mann-Whitney U test). (C) Biaxial flow cytometry plot showing expression of CD4 and CXCR5 on cells gated for CD4⁺CD45RA^-FOXP3^hi. This representative example from a patient with lymphoma shows cTfr 21.4% of CD4⁺CD45RA^-FOXP3^hi cells. (D) cTfr as a proportion of total CD4⁺ T-cells. Horizontal lines represent the median and bars represent inter-quartile range. There are no significant differences between normal subjects (n = 11) and patient with MZL (n = 4) or BNHL (n = 9). (E) Biaxial flow cytometry plot showing expression of CD4 and FOXP3 on cells gated for CD4⁺CD45RA^-CXCR5⁺. Gates were set by employing human tonsillar T-cells as controls. This representative example from a patient with lymphoma shows cTfr (FOXP3^hi) 7.3% and cTfh (FOXP3^-/lo) 92.7% of CD4⁺CD45RA^-CXCR5⁺ cells. (F) cTfr:cTfh ratio. Horizontal lines represent the median and bars represent inter-quartile range. BNHL patients (n = 9) showed a significant difference (Mann-Whitney U-test, P = 0.038) from normal subjects (n = 11) but MZL patients (n = 4) did not.