Science Advances

Supplementary Materials

This PDF file includes:

  • Materials and Methods
  • Table S1. Summary of analytical data of (80 nm) BP nanoparticles.
  • Table S2. Summary of analytical data of all BNF-HER nanoparticles prepared.
  • Table S3. Characteristics of breast cancer cell lines used in the study.
  • Table S4. Summary of analytical data of BNF-IgG nanoparticles.
  • Table S5. Summary of analytical data of BNF-HER nanoparticles that passed in vitro qualification testing.
  • Table S6. Summary of immune modifications in mouse strains used for study.
  • Table S7. Summary of Aperio imaging settings used for digital analysis of tissue sections.
  • Table S8. Definitions of parameters used for Aperio imaging settings.
  • Table S9. Antibodies used for flow cytometry and their dilutions.
  • Table S10. Summary of numbers and strains of mice used in the study.
  • Table S11. Summary of one-factor model statistical analysis of iron measurements in xenograft models.
  • Table S12. Summary of two-factor model statistical analysis of iron measurements in xenograft models.
  • Table S13. Summary of three-factor model statistical analysis of iron measurements in xenograft models.
  • Table S14. Summary of one-factor model statistical analysis of Prussian blue histopathology analyses in xenograft models.
  • Table S15. Summary of two-factor model statistical analysis of Prussian blue histopathology analyses in xenograft models.
  • Table S16. Summary of three-factor model statistical analysis of Prussian blue histopathology analyses in xenograft models.
  • Table S17. Summary of statistical analysis of whole tumor digests flow cytometry in huHER2 allograft model.
  • Table S18. Summary of statistical analysis of nanoparticle-associated fractions (magnetic-sorted sediment) from flow cytometry in huHER2 allograft model.
  • Table S19. Summary of statistical analysis of nanoparticle-depleted fractions (magnetic-sorted supernatant) from flow cytometry in huHER2 allograft model.
  • Table S20. Summary of statistical analysis of iron measurements (ICP-MS) obtained from the livers of xenograft models.
  • Table S21. Ratio of Fe level between groups (treatment).
  • Table S22. Ratio of Fe level between groups (strains).
  • Table S23. Statistical analysis of ICP-MS huHER2-FVB/N lymph node data.
  • Table S24. Statistical analysis of ICP-MS huHER2-FVB/N spleen data.
  • Table S25. Statistical analysis of ICP-MS huHER2-FVB/N liver data.
  • Table S26. Ratio of percent positive between groups.
  • Table S27. Statistical analysis of tumor weight in huHER2-FVB/N.
  • Table S28. Statistical analysis of tumor growth in huHER2-FVB/N.
  • Table S29. Statistical analysis of whole tumor flow data third day.
  • Table S30. Statistical analysis of whole tumor flow data seventh day.
  • Table S31. Statistical analysis of whole tumor flow data 14th day.
  • Table S32. Statistical analysis of tumor weight–huHER2 allograft in nude mice.
  • Table S33. Statistical analysis of tumor growth–huHER2 allograft in nude mice (from initial day to 21st day).
  • Fig. S1. Representative images showing immunofluorescence staining of BH particles.
  • Fig. S2. Subtracting endogenous iron using PBS controls reveals little tumor retention of plain nanoparticles, and retention of BH nanoparticles is independent of tumor expression of the target antigen HER2.
  • Fig. S3. Retention of Herceptin-labeled BNF nanoparticles by xenograft tumors depends on immune strain of host.
  • Fig. S4. Weak correlations were found between deposits of plain nanoparticles and HER2, CD31+, or IBA-1+ regions in tumors of mice injected with BP nanoparticles.
  • Fig. S5. BNF nanoparticles labeled with a nonspecific IgG polyclonal human antibody were retained by tumors.
  • Fig. S6. Histopathology data support ICP-MS results for tumor retention of nanoparticles, and ICP-MS data show nanoparticles accumulated in lymph nodes, spleens, and livers of injected mice.
  • Fig. S7. Within tumors, nanoparticles localized in stromal regions rather than in cancer cell–rich regions.
  • Fig. S8. Gating for flow cytometry was conducted to ascertain immune cell populations residing in tumors.
  • Fig. S9. Flow cytometry analysis of huHER2 tumors harvested from immune competent mice reveals tumor immune microenvironment changes, and magnetically sorted tumor immune cell populations demonstrates impact of nanoparticles on tumor immune cells in response to intravenous nanoparticle delivery.
  • Fig. S10. Pan-leukocyte inhibition abrogates BH nanoparticle retention in tumors.
  • Fig. S11. Systemic exposure to BNF nanoparticles resulted in tumor growth inhibition but only if the host has an intact (adaptive) immune system (i.e., T cells).
  • Fig. S12. Following systemic exposure to nanoparticles, intratumor T cell populations decline through the third day and then increase by day 7 relative to PBS controls.
  • Fig. S13. Exposure to nanoparticles induces changes in adaptive immune signaling in tumors of nanoparticle-treated mice.
  • Fig. S14. Changes in innate cell population in tumors of nanoparticle-treated mice.
  • Fig. S15. Data suggest that systemically delivered BNF nanoparticles are preferentially sequestered by inflammatory immune cells within the TME, resulting in immune recognition of the tumor.

Download PDF

Files in this Data Supplement: