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Blood, Vol. 113, Issue 15, 3397-3405, April 9, 2009
A peptomimetic inhibitor of BCL6 with potent antilymphoma effects in vitro and in vivo
Blood Cerchietti et al.
113: 3397
Supplemental materials for: Cerchietti et al
Files in this Data Supplement:
- Document 1. Supplemental materials and methods (PDF, 110 KB)
- Figure S1. Fusogenic motif enhances the biological activity of BPI (JPG, 72.1 KB)
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(A) An example showing the effect on viability of control peptides alone and mutant peptides of the BPI S1, S2, S3, and S6.2 (mS1, mS2, mS3, and mS6.2) in a BCL6-dependent (Ly10) and a BCL6 independent (Ly4) cell line. (B) Effect of different peptide design (X-axis) on DLBCL viability (Y-axis). The fusogenic peptide (Fu) alone or in combination with a TAT peptide had little or no effect on cells. Co-administration of the S2-BPI with Fu significantly enhances its anti-lymphoma activity in BCL6-dependent DLBCL cells (Ly1, Ly7, and Ly10) but not in the BCL6 independent Ly4 cell line. A mass-equivalent dose of the combined Fu-TAT-GRSIHEIPR (S3) peptide was similar in efficacy to the concomitant administration of S2 plus Fu. All the peptides were administered every 4 h during a 48 h interval.
- Figure S2. RI-BPI affects the formation of germinal centers in immunized mice (JPG, 115 KB)
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Sheep red blood cells were administered intraperitoneally in 10 C57BL/6 mice to induce a T-cell–dependent immune response and generate germinal centers. On day 3, the mice were randomized in 2 groups (n=5 per group) and treated intraperitoneally with 500 µg of RI-BPI or control peptide per day. After 7 days of injections the animals were sacrificed and spleens were examined for formation of germinal centers by peanut agglutinin (PNA) histochemistry. (A) Representative images of spleens from control and treated mice showing PNA+ (activated GC B cells, upper panel) and B220+ (total GC B cells, lower panel) clusters in brown. Hematoxilin was used as counterstaining (blue). (B) Number (left panel) and size (right panel) of PNA+ clusters from (A). PNA clusters were counted and measured in each whole-spleen digitally reconstituted longitudinal sections, and data represent the median of 5 mice with SD. Images were processed using Image J software. P values were obtained with T-test. LPF: Low power field (40× final magnification).
- Figure S3. Human β2-microglobulin levels are closely associated with tumor burden in mice baring DLBCL xenografts (JPG, 43.2 KB)
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Pearson’s correlation of serum levels of human β2-microglobulin (Y-axis) and tumor burden in grams (X-axis) determined at day 10 in SUDHL4 mice.
- Figure S4. RI-BPI is non-toxic in mice exposed for periods up to 52 weeks (JPG, 193 KB)
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Representative images of hematoxilin-eosin stained tissues harvested from mice treated with control peptide (first column), RI-BPI 500 µg/day × 21 days (second column) and RI-BPI 500 µg/week × 52 weeks (third column). Anti-RI-BPI antibodies were not detected in the serum of these mice.
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