Journal: Molecular pharmaceutics

Article Title: CD19-Targeted Nanodelivery of Doxorubicin Enhances Therapeutic Efficacy in B-Cell Acute Lymphoblastic Leukemia.

doi: 10.1021/acs.molpharmaceut.5b00071

Figure Lengend Snippet: Figure 2. Quantification of CD19 total and surface expression levels in REH and RS4;11 B-ALL cells: (A and B) Immunoblot and quantification of the CD19 levels. (C) Flow cytometry analysis of surface levels of CD19 (Note FITC-(anti-CD19 Ab) control indicated by black arrow.) Quantified data shown are averages of two independent experiments. Statistical significance was determined using a two-tailed Student’s t test where a p value < 0.05 was considered significant.

Article Snippet: Human acute leukemia cell lines RS4;11 (ATCCCRL-1873, established from a patient with B-ALL at first relapse) and REH (ATCC CRL-8286, also established from a patient with B-ALL at first relapse) were purchased from the American Type Culture Collection (ATCC, Manassas, VA).

Techniques: Expressing, Western Blot, Flow Cytometry, Control, Two Tailed Test

Journal: Molecular pharmaceutics

Article Title: CD19-Targeted Nanodelivery of Doxorubicin Enhances Therapeutic Efficacy in B-Cell Acute Lymphoblastic Leukemia.

doi: 10.1021/acs.molpharmaceut.5b00071

Figure Lengend Snippet: Figure 3. CD19-specific targeting and uptake of anti-CD19-Ab-conjugated NPs. (A) CLSM analysis to compare uptake of CD19-NR-NPs and NR-NPs (final concentration ≈1 μM of encapsulated DOX) in REH and RS4;11 cells for 1 h at 37 °C. The uptake pattern was similar at 5 min. Note that even after 1 h exposure REH cells show minimal uptake. Scale bar = 10 μm. (B) Quantification of cell uptake levels of CD19-NR-NPs and NR-NPs in REH and RS4;11 cells. (C) CLSM analysis to compare uptake of CD19-NR-NPs with isotype-matched nonspecific uptake of IgG-NR-NPs (final concentration ≈ 1 μM ofencapsulated DOX) inRS4;11 cells for 1 h at 37°C. Scalebar = 7.5 μm.(D). CLSM analysisto confirm CD19-dependent uptake ofCD19-NR-NPs (final concentration ≈1 μM of encapsulated DOX) in RS4;11 cells for 1 h at 37 °C. Scale bar = 10 μm. Quantified data shown are averages of three independent experiments and are indicated as mean ± SD. Statistical significance was determined using the Holm−Šidák method, with α = 5.000%.

Article Snippet: Human acute leukemia cell lines RS4;11 (ATCCCRL-1873, established from a patient with B-ALL at first relapse) and REH (ATCC CRL-8286, also established from a patient with B-ALL at first relapse) were purchased from the American Type Culture Collection (ATCC, Manassas, VA).

Techniques: Concentration Assay

Journal: Molecular pharmaceutics

Article Title: CD19-Targeted Nanodelivery of Doxorubicin Enhances Therapeutic Efficacy in B-Cell Acute Lymphoblastic Leukemia.

doi: 10.1021/acs.molpharmaceut.5b00071

Figure Lengend Snippet: Figure 5. CD19-mediated induction of apoptosis in leukemia cells. (A) Comparison of toxicity induced by free DOX and CD19-DOX-NPs (≈100 nM DOX) in REH and RS4;11 cells. (B) Competition assay to confirm CD19-receptor-dependent induction of toxicity by CD19-DOX-NPs (≈100 nM DOX) in RS4;11 cells. (C) Dose-dependent cleaved PARP levels caused by treatment of CD19-DOX-NPs with RS4;11 cells (48 h). Quantified data shown are averages of three independent experiments. Statistical significance was determined using the Holm−Šidák method, with α = 5.000%.

Article Snippet: Human acute leukemia cell lines RS4;11 (ATCCCRL-1873, established from a patient with B-ALL at first relapse) and REH (ATCC CRL-8286, also established from a patient with B-ALL at first relapse) were purchased from the American Type Culture Collection (ATCC, Manassas, VA).

Techniques: Comparison, Competitive Binding Assay

Journal: Molecular pharmaceutics

Article Title: CD19-Targeted Nanodelivery of Doxorubicin Enhances Therapeutic Efficacy in B-Cell Acute Lymphoblastic Leukemia.

doi: 10.1021/acs.molpharmaceut.5b00071

Figure Lengend Snippet: Figure 6. CD19-targeting increases the cell-associated DOX (intracellular and cell-surface-bound, caDOX) levels in RS4;11 cells: (A) HPLC/MS/MS representative quantitation of caDOX levels in REH and RS4;11 cells treated with CD19-DOX-NPs (≈1 μM DOX, for 1 h at 37 °C). (B) REH and RS4;11 caDOX levels on treatment with free DOX for 1 h at 37 °C. (C) REH and RS4;11 caDOX levels on treatment with CD19-DOX-NPs for 1 h at 37 °C. Quantified data shown are averages of three independent experiments. Statistical significance was determined using the Holm−Šidák method, with α = 5.000%.

Article Snippet: Human acute leukemia cell lines RS4;11 (ATCCCRL-1873, established from a patient with B-ALL at first relapse) and REH (ATCC CRL-8286, also established from a patient with B-ALL at first relapse) were purchased from the American Type Culture Collection (ATCC, Manassas, VA).

Techniques: Tandem Mass Spectroscopy, Quantitation Assay