Skip to main content
SpringerLink
Log in

Preparation and evaluation of norcantharidin-encapsulated liposomes modified with a novel CD19 monoclonal antibody 2E8

  • Published:
Journal of Huazhong University of Science and Technology [Medical Sciences] Aims and scope Submit manuscript

Summary

In this study, norcanthridin (NCTD)-encapsulated liposomes were modified with a novel murine anti-human CD19 monoclonal antibody 2E8 (2E8-NCTD-liposomes) and the targeting efficiency and specific cytotoxicity of 2E8-NCTD-liposomes to CD19+ leukemia cells were evaluated. BALB/c mice were injected with 2E8 hybridoma cells to obtain 2E8 monoclonal antibody (mAb). NCTD-liposomes were prepared by using film dispersion method. 2E8 mAbs were linked to NCTD-liposomes using post-incorporation technology. Flow cytometry showed that the targeting efficiency of purified 2E8 mAbs on CD19+ Nalm-6 cells was 99.93%. The purified 2E8 mAbs were conjugated with NCTD-liposomes to prepare 2E8-NCTD-liposomes whose targeting efficiency on CD19+ Nalm-6 was also 95.82%. The average size of 2E8-NCTD-liposomes was 118.32 nm in diameter. HPLC showed that the encapsulation efficiency of NCTD was 46.51%. When the molar ratio of 2E8/Mal-PEG2000-DSPE reached 1:50, we obtained the liposomes with 9 2E8 molecules per liposome. The targeting efficiency of 2E8-NCTD-liposomes on CD19+ leukemia cells was significantly higher than that on CD19-leukemia cells. Similarly, the targeting efficiency of the immunoliposomes was also higher than that of the NCTD-liposomes on CD19+ leukemia cells. Those results were consistent with those observed by laser scanning confocal microscopy. 3-(4, 5-Dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) assay demonstrated that 2E8-NCTD-liposomes specifically killed Nalm-6 cells in a dose- and time-dependent manner. The viability of Nalm-6 cells treated by 2E8-NCTD-liposomes was significantly lower than that of Molt-3 cells and it was also significantly lower than that of Nalm-6 cells treated with the same concentration of NCTD-liposomes or free NCTD. We are led to concluded that 2E8 antigen can serve as a specific targeting molecule of B lineage hematopoietic malignancies for liposome targeting, and 2E8-NCTD-liposomes can be used as a new and effective means for the treatment of B lineage hematopoietic malignancies.

This is a preview of subscription content, log in via an institution to check access.

Access this article

Log in via an institution

Price excludes VAT (USA)
Tax calculation will be finalised during checkout.

Instant access to the full article PDF.

Institutional subscriptions

Similar content being viewed by others

References

  1. Batist G, Barton J, Chaikin P, et al. Myocet (liposome-encapsulated doxorubicin citrate): a new approach in breast cancer therapy. Expert Opin Pharmacother, 2002, 3(12):173917–173951

    Article  Google Scholar 

  2. Cabanes A, Briggs KE, Gokhale PC, et al. Comparative in vivo studies with paclitaxel and liposome-encapsulated paclitaxel. Int J Oncol, 1998,12(5):1035–1s040

    CAS  PubMed  Google Scholar 

  3. Gao Z, Lukyanov AN, Chakilam AR, et al. PEG-PE/phosphatidylcholine mixed immunomicelles specifically deliver encapsulated taxol to tumor cells of different origin and promote their efficient killing. J Drug Target, 2003,11(2):87–92

    Article  CAS  PubMed  Google Scholar 

  4. Park JW, Benz CC, Martin FJ. Future directions of liposome- and immunoliposome-based cancer therapeutics. Semin Oncol, 2004,31(6 Suppl 13):196–205

    Article  CAS  PubMed  Google Scholar 

  5. Uckun FM, Jaszcz W, Ambrus JL, et al. Detailed studies on expression and function of CD19 surface determinant by using B43 monoclonal antibody and the clinical potential of anti-CD19 immunotoxins. Blood, 1988,71(1): 13–29

    CAS  PubMed  Google Scholar 

  6. Sapra P, Allen TM. Internalizing antibodies are necessary for improved therapeutic efficacy of antibody-targeted liposomal drugs. Cancer Res, 2002, 62(24):7190–9194

    CAS  PubMed  Google Scholar 

  7. Sapra P, Allen TM. Improved outcome when B-cell lymphoma is treated with combinations of immunoliposomal anticancer drugs targeted to both the CD19 and CD20 epitopes. Clin Cancer Res, 2004,10(7): 2530–2537

    Article  CAS  PubMed  Google Scholar 

  8. Harata M, Soda Y, Tani K, et al. CD19-targeting liposomes containing imatinib efficiently kill Philadelphia chromosome-positive acute lymphoblastic leukemia cells. Blood, 2004,104(5):1442–1449

    Article  CAS  PubMed  Google Scholar 

  9. Cheng WW, Allen TM. Targeted delivery of anti-CD19 liposomal doxorubicin in B-cell lymphoma: a comparison of whole monoclonal antibody, Fab’ fragments and single chain Fv. J Control Release, 2008,126(1):50–58

    Article  CAS  PubMed  Google Scholar 

  10. Laginha K, Mumbengegwi D, Allen T. Liposomes targeted via two different antibodies: assay, B-cell binding and cytotoxicity. Biochim Biophys Acta, 2005,1711(1): 25–32

    Article  CAS  PubMed  Google Scholar 

  11. Zhang JY, Tang Y, Shen H, et al. Targeting and internalization of sterically stabilized liposome modified with ZCH-4-2E8. J Huazhong Univ Sci Technol [Med Sci], 2009,29(3):273–280

    Article  CAS  Google Scholar 

  12. Liu D, Chen Z. The effects of cantharidin and cantharidin derivates on tumour cells. Anticancer Agents Med Chem, 2009,9(4):392–396

    CAS  PubMed  Google Scholar 

  13. Krautheim A, Brechlin P, Becker K, et al. Hamster pancreatic beta cell lines with altered sensitivity towards apoptotic signalling by phosphatase inhibitors. Br J Pharmacol, 2000,129(4):687–694

    Article  CAS  PubMed  Google Scholar 

  14. McCluskey A, Ackland SP, Bowyer MC, et al. Cantharidin analogues: synthesis and evaluation of growth inhibition in a panel of selected tumour cell lines. Bioorg Chem, 2003,31(1):68–79

    Article  CAS  PubMed  Google Scholar 

  15. Wera S, Hemmings BA. Serine/threonine protein phosphatases. Biochem J, 1995,311(Pt 1):17–29

    CAS  PubMed  Google Scholar 

  16. Dorn DC, Kou CA, Png KJ, et al. The effect of cantharidins on leukemic stem cells. Int J Cancer, 2009,124(9):2186–2199

    Article  CAS  PubMed  Google Scholar 

  17. Wu KD, Cho YS, Katz J, et al. Investigation of antitumor effects of synthetic epothilone analogs in human myeloma models in vitro and in vivo. Proc Natl Acad Sci USA, 2005,102(30):10640–10645

    Article  CAS  PubMed  Google Scholar 

  18. Li LX, Tang YM, Zhang HZ, et al. Preparation of the immunotoxin 2E8-norcantharidin and its targeting killing effect in vitro. Zhonghua Er Ke Za Zhi (Chinese), 2008,46(7):493–497

    Google Scholar 

  19. Tang Y, Scollard D, Chen P, et al. Imaging of HER2/neu expression in BT-474 human breast cancer xenografts in athymic mice using [(99m)Tc]-HYNIC-trastuzumab (Herceptin) Fab fragments. Nucl Med Commun, 2005, 26(5):427–432

    Article  CAS  PubMed  Google Scholar 

  20. Sapra P, Moase EH, Ma J, et al. Improved therapeutic responses in a xenograft model of human B lymphoma (Namalwa) for liposomal vincristine versus liposomal doxorubicin targeted via anti-CD19 IgG2a or Fab’ fragments. Clin Cancer Res, 2004,10(3):1100–1111

    Article  CAS  PubMed  Google Scholar 

  21. Allen TM, Mumbengegwi DR, Charrois GJ. Anti-CD19-targeted liposomal doxorubicin improves the therapeutic efficacy in murine B-cell lymphoma and ameliorates the toxicity of liposomes with varying drug release rates. Clin Cancer Res, 2005,11(9):3567–3573

    Article  CAS  PubMed  Google Scholar 

  22. Iden DL, Allen TM. In vitro and in vivo comparison of immunoliposomes made by conventional coupling techniques with those made by a new post-insertion approach. Biochim Biophys Acta, 2001,1513(2):207–216

    Article  CAS  PubMed  Google Scholar 

  23. Zhou Y, Drummond DC, Zou H, et al. Impact of single-chain Fv antibody fragment affinity on nanoparticle targeting of epidermal growth factor receptor-expressing tumor cells. J Mol Biol, 2007,371(4):934–947

    Article  CAS  PubMed  Google Scholar 

  24. Sapra P, Tyagi P, Allen TM. Ligand-targeted liposomes for cancer treatment. Curr Drug Deliv. 2005. 2(4):369–381

    Article  CAS  PubMed  Google Scholar 

  25. Allen TM. Ligand-targeted therapeutics in anticancer therapy. Nat Rev Cancer. 2002. 2(10): 750–63.

    Article  CAS  PubMed  Google Scholar 

  26. Sato S, Tedder TF. CD19 Workshop Panel report. Leucocyte Typing IV. Edited by Tadamitsu Kishimoto, et al. Garland Publishing, Inc. New York & London: 1997. 133–134

    Google Scholar 

  27. Wu JM, Ren TQ. Preparation and characterization of encapsulated norcantharidin in liposomes. Chin Pharm J, 2005,40(19):1485–1489

    CAS  Google Scholar 

  28. Yang T, Choi MK, Cui FD, et al. Preparation and evaluation of paclitaxel-loaded PEGylated immunoliposome. J Control Release, 2007,120(3):169–177

    Article  CAS  PubMed  Google Scholar 

  29. Maruyama K, Takizawa T, Yuda T, et al. Target ability of novel immunoliposomes modified with amphipathic poly(ethylene glycol)s conjugated at their distal terminals to monoclonal antibodies. Biochim Biophys Acta, 1995, 1234(1):74–80

    Article  PubMed  Google Scholar 

  30. Xu Y, Zhu Z, Wu K, et al. Construction and identification of NF-κB promotor regulatory eukaryotic bicistronic expression vector pNF-κB-IRES2-EGPF-p27. Acta Med Univ Sci Technol Huazhong (Chinese), 2008,37(6): 708–711

    CAS  Google Scholar 

  31. Lu J, Jeon E, Lee BS, et al. Targeted drug delivery crossing cytoplasmic membranes of intended cells via ligand-grafted sterically stabilized liposomes. J Control Release, 2006,110(3):505–513

    Article  CAS  PubMed  Google Scholar 

Download references

Author information

Authors and Affiliations

  1. Division of Hematology-Oncology, Children’s Hospital, School of Medicine, Zhejiang University, Hangzhou, 310003, China

    Jingying Zhang  (张晶樱), Yongmin Tang  (汤永民), Baiqin Qian  (钱柏芹) & Hongqiang Sheng  (沈红强)

Authors
  1. Jingying Zhang  (张晶樱)
    View author publications

    You can also search for this author in PubMed Google Scholar

  2. Yongmin Tang  (汤永民)
    View author publications

    You can also search for this author in PubMed Google Scholar

  3. Baiqin Qian  (钱柏芹)
    View author publications

    You can also search for this author in PubMed Google Scholar

  4. Hongqiang Sheng  (沈红强)
    View author publications

    You can also search for this author in PubMed Google Scholar

Corresponding author

Correspondence to Yongmin Tang  (汤永民).

Additional information

This project was supported by a grant from the National Natural Sciences Foundation of China (No. 30170391), the Zhejiang Provincial Fund of Natural Sciences (No: Z205166) and the Zhejiang Provincial Fund of Science and Technology Bureau (No: 2007C23007).

Rights and permissions

Reprints and permissions

About this article

Cite this article

Zhang, J., Tang, Y., Qian, B. et al. Preparation and evaluation of norcantharidin-encapsulated liposomes modified with a novel CD19 monoclonal antibody 2E8. J. Huazhong Univ. Sci. Technol. [Med. Sci.] 30, 240–247 (2010). https://doi.org/10.1007/s11596-010-0222-1

Download citation

  • Received:

  • Published:

  • Issue Date:

  • DOI: https://doi.org/10.1007/s11596-010-0222-1

Key words

Search

Navigation