Double staining of PD-L1 (1:50; clone E1L3N; Cell Signaling, Danvers,
MA) or PD-L2 (1:50; clone D7U8C; Cell Signaling) and PAX5 (1:100; clone 24; BD
Biosciences, San Jose, CA) was performed with an automated staining system (Bond
III; Leica Biosystems, Buffalo Grove, IL) as previously described (Ansell et al., 2015 (link)). Stained slides were
scored by an expert dermatopathologist (JG) and expert hematopathologist (AB),
and percentages of both tumor PDL-1 or PDL-2 and microenvironment PDL-1 or PDL-2
were calculated by scoring 100–200 cells in each category. The threshold
for PD-L1 and PD-L2 expression was defined at 30% for PAX5-positive tumor cells
and at 20% for PAX5-negative immune cell microenvironment, as reported by others
(Kiyasu et al., 2015 (link)).
MA) or PD-L2 (1:50; clone D7U8C; Cell Signaling) and PAX5 (1:100; clone 24; BD
Biosciences, San Jose, CA) was performed with an automated staining system (Bond
III; Leica Biosystems, Buffalo Grove, IL) as previously described (Ansell et al., 2015 (link)). Stained slides were
scored by an expert dermatopathologist (JG) and expert hematopathologist (AB),
and percentages of both tumor PDL-1 or PDL-2 and microenvironment PDL-1 or PDL-2
were calculated by scoring 100–200 cells in each category. The threshold
for PD-L1 and PD-L2 expression was defined at 30% for PAX5-positive tumor cells
and at 20% for PAX5-negative immune cell microenvironment, as reported by others
(Kiyasu et al., 2015 (link)).