Legends
Figure 1: Patients with limited disease show higher serum rituximab levels. Serum rituximab levels of the B-NHL cohort at day five after treatment. Rituximab levels (µg/ml) of patients, grouped after the B-NHL stratification criteria in R1/R2 (n = 6) and R3/R4 (n = 8), were measured with an ELISA at day five after treatment. Patients with limited disease (risk groups R1 and R2) presented with a significantly higher median serum rituximab level of 250 µg/ml than patients with advanced disease (risk groups R3 and R4) with 40 µg/ml (p = 0.0001). An unpaired two-tailed student’s t-test was performed.
Figure 2: Flow cytometric analysis of FcγR expression in monocyte subtypes before (A) and at day five after (B) rituximab treatment. FcγR expression in classical (cl, CD14+CD16-), intermediate (int, CD14+CD16+) and non-classical (non-cl, CD14lowCD16+) monocytes was analyzed. FcγRI expression was significantly increased in classical monocytes (p = 0.002). FcγRII expression was significantly decreased in classical and intermediate monocytes, while FcγRIII expression was not influenced by rituximab treatment (p = 0.023 and p = 0.006). Samples were measured in duplicates (n = 10). Expression levels are displayed as median fluorescent intensity (MFI). A paired two-tailed student’s t-test was performed.
Figure 3: S100A9 and S100A12 expression is increased in monocyte subtypes at day five after rituximab treatment while serum levels are decreased. A S100A9 and S100A12 expression was measured via flow cytometry before (A) and at day five after (B) rituximab treatment in classical (cl, CD14+CD16-), intermediate (int, CD14+CD16+) and non-classical (non-cl, CD14lowCD16+) monocytes. S100A9 expression significantly increased after rituximab treatment in all monocyte subtypes (p = 0.005, p = 0.009 and p = 0.031). S100A12 expression significantly increased in classical and intermediate monocytes after rituximab treatment (p= 0.037 and p = 0.024). Expression levels are displayed as median fluorescent intensity (MFI). A paired two-tailed student’s t-test was performed (n = 10). B Serum S100A8/9 and S100A12 levels were measured with an ELISA before (A) and at day five after (B) rituximab treatment. Conversely to expression levels, serum S100A8/9 and S100A12 levels significantly decreased after treatment (p = 0.021 andp = 0.012). Values are displayed in ng/ml. A paired two-tailed student’s t-test was performed (n = 10).
Figure 4: CD57, PRF1 and GrzB expression is decreased in NK cells at day 5 after rituximab treatment. CD57, perforin (PRF1) and granzyme B (GrzB) expression was measured via flow cytometry before (A) and at day five after (B) rituximab treatment in NK cells. CD57 (p = 0.006), PRF1 (p = 0.045) and GrzB (p = 0.028) expression were significantly decreased at day five after rituximab treatment in NK cells. Expression levels are displayed as median fluorescent intensity (MFI) in CD56+ positive cells. A paired two-tailed student’s t-test was performed (n = 12).