Figure 1.
Pharmacokinetics of serum rituximab in cynomolgus monkeys.
PK analysis of serum rituximab levels after (A) a single SC dose of rituximab preformulated in rHuPH20 administered at 20 mg/kg (Individual animal samples with mean score shown (n=3)), and (B) 2 × 10 mg/kg doses of SC (preformulated in rHuPH20) or standard IV rituximab, given 7 days apart (Individuals animal samples with mean shown (n=4)).
Figure 2.
CD20 target coverage in B-cells and according to CD21+ status.
Paradigms for flow cytometry staining: (A) CD20-independent identification of B cells using fsc/scc lymphocyte gating followed by CD4/CD3 negative gating and CD40 positive CD16 negative gating shows a progressively increasing specific B cell population; (B) Free surface CD20 levels on B cells (as identified in A) with and without rituximab treatment to show target coverage of CD20 compared to CD4+/CD3+ T cells; (C) Identification of CD21+ and CD21- peripheral blood B-cell subsets (within the CD4-/CD3-/CD16-/CD40+ gated B cells as identified in A) showing CD21, CD40 and CD20 levels of the subsets.
Figure 3.
CD20 target coverage and B-cell depletion in lymph nodes.
Lymph node analysis of (A) CD20 target coverage as determined by flow cytometric staining for free surface CD20 levels on (B cells identified as in Figure 2A) and (B) depletion of those lymph node B-cellsat baseline and 9 days after second dose of subcutaneous or intravenous rituximab as determined by a ratio to CD4+/CD3+ T cells (Individuals animal samples with mean shown (n=4 rituximab treated groups, n=3 PBS vehicle treated groups)).
Figure 4.
CD20 target coverage and B-cell depletion in Peripheral Blood.
PBMC analysis of (A) CD20 target coverage as determined by flow cytometric staining for free surface CD20 levels on (B cells identified as in Figure 2A) and (B) depletion of those PBMC B-cells at baseline and 2, 9, and 14 days after second dose of subcutaneous or intravenous rituximab as determined by a ratio to CD4+/CD3+ T cells (Individuals animal samples with mean shown (n=4 rituximab treated groups, n=3 PBS vehicle treated groups)); and long-term PBMC analysis of (C) percent free surface CD20 target coverage and (D) percent remaining B-cells out to 63 days; normalized to PBS vehicle (group mean ± SD (n=4 rituximab treated groups, n=3 PBS vehicle treated groups)) with ex-vivo rituximab treatment of PBS vehicle sample to show maximal target coverage.
Figure 5.
Depletion of Peripheral Blood CD21+ and CD21- B-cell subsets.
Short and long-term analysis of PBMC B-cell depletion of (A) CD21+ and (B) CD21- B-cell subsets (as identified in Figure 2C). Individuals animal samples with mean shown for baseline, day 2, day 9 and day 14 post second dose (top) and percent remaining as normalized to PBS vehicle treated group (bottom) (group mean ± SD (n=4 rituximab treated groups, n=3 PBS vehicle treated groups).