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. 2019 Sep 19;105(6):1593–1603. doi: 10.3324/haematol.2019.220160

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Figure 3. — Ki67pos cells are in close proximity to follicular helper T cells (TFH) in follicular lymphoma lymph nodes. (A1) Representative image of a neoplastic follicle showing Ki67pos cells (green) in close proximity to CD4^pos (red), PD-1^Hi (white) T cells. The scale bar represents 25 μm. (A2) Binary image of (A1), the binary layers of Ki67 (green) and the CD4-PD-1^Hi intersection (magenta) are shown highlighting the close association of Ki67^pos cells to PD-1^Hi T cells. (B1) Representative image demonstrating that the majority of the PD-1^Hi cells in contact with Ki67pos cells (red) are also positive for ICOS (green). (B2) This is highlighted in the binary layer 3D reconstruction of the same image, PD-1/ICOS^pos (magenta) and Ki67 (green). Images representative of n=100 images from n=23 follicular lymphoma (FL) samples (4A), n=43 images from n=13 samples (4B). (C1) Ki67=blue, CD20=red, PD-1=white, CD3=green. Low power image (x10) showing Ki67^pos and Ki67^neg CD20^pos B-cell co-localisation with PD1^Hi CD3^pos T cells in FL. Within the follicles there are areas of low proliferation (low Ki67=blue) where there are few PD1^Hi (white) CD3^pos T cells (green) - area highlighted by yellow oval, whereas in areas where there is high Ki67, there are more PD1^Hi, CD3pos T cells (area highlighted by white circle) and they are frequently in contact with Ki67pos CD20pos FL B cells. Scale bar represents 100 μm. (C2) High power image (x60) in which the close correlation of Ki67pos (blue) B cells with PD1^Hi (white) CD3^pos (green) cells can be seen, whilst the CD20^pos (red), Ki67^neg cells are less frequently in contact with follicular helper T cells (TF_H). Scale bar represents 50 μm. (C3) contingency tables showing that Ki67pos B cells are significantly more likely to be in contact with TF_H than Ki67^neg B-cells in FL (for all samples analyzed together [n=25 images from n=7 follicular lymphoma specimens] χ² 595, P<0.0001).