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Heat-Shock Protein 70 as a Tumor Antigen for in vitro Dendritic Cell Pulsing in Renal Cell Carcinoma Cases

  • Meng, Fan-Dong (The Second Lab of Cancer Research Institute, The First Hospital of China Medical University) ;
  • Sui, Cheng-Guang (The Second Lab of Cancer Research Institute, The First Hospital of China Medical University) ;
  • Tian, Xin (The Second Lab of Cancer Research Institute, The First Hospital of China Medical University) ;
  • Li, Yan (The Second Lab of Cancer Research Institute, The First Hospital of China Medical University) ;
  • Yang, Chun-Ming (Department of Urology, The First Hospital of China Medical University) ;
  • Ma, Ping (The Second Lab of Cancer Research Institute, The First Hospital of China Medical University) ;
  • Liu, Yun-Peng (Department of Medical Oncology, The First Hospital of China Medical University) ;
  • Jiang, You-Hong (The Second Lab of Cancer Research Institute, The First Hospital of China Medical University)
  • Published : 2014.11.06

Abstract

Immunological functions of heat shock proteins (HSPs) have long been recognized. In this study we aimed to efficiently purify HSP70 from renal cell carcinoma and test it as a tumor antigen for pulsing dendritic cells in vitro. HSP70 was purified from renal cell carcinoma specimens by serial column chromatography on Con A-sepharose, PD-10, ADP-agarose and DEAE-cellulose, and finally subjected to fast protein liquid chromatography (FPLC). Dendritic cells derived from the adherent fraction of peripheral blood mononuclear cells were cultured in the presence of IL-4 and GM-CSF and exposed to tumor HSP70. After 24 hours, dendritic cells were phenotypically characterized by flow cytometry. T cells obtained from the non-adherent fraction of peripheral blood mononuclear cells were then co-cultured with HSP70-pulsed dendritic cells and after 3 days T cell cytotoxicity towards primary cultured renal cell carcinoma cells was examined by Cell Counting Kit-8 assay. Dendritic cells pulsed in vitro with tumor-derived HSP70 expressed higher levels of CD83, CD80, CD86 and HLA-DR maturation markers than those pulsed with tumor cell lysate and comparable to that of dendritic cells pulsed with tumor cell lysate plus TNF-${\alpha}$. Concomitantly, cytotoxic T-lymphocytes induced by HSP70-pulsed dendritic cells presented the highest cytotoxic activity. There were no significant differences when using homologous or autologous HSP70 as the tumor antigen. HSP70 can be efficiently purified by chromatography and induces in vitro dendritic cell maturation in the absence of TNF-${\alpha}$. Conspecific HSP70 may effectively be used as a tumor antigen to pulse dendritic cells in vitro.

Keywords

Heat-shock protein 70;dendritic cells;renal cell carcinoma;tumor antigen

References

  1. Besch GJ, Wolberg WH, Gilchrist KW, et al (1983). A comparison of methods for the production of monodispersed cell suspensions from human primary breast carcinomas. Breast Cancer Res Treat, 3, 15-22. https://doi.org/10.1007/BF01806230
  2. Amato R. J. (2008). Vaccine therapy for renal cancer. Expert Rev Vaccines, 7, 925-35. https://doi.org/10.1586/14760584.7.7.925
  3. Bellone S, Pecorelli S, Cannon MJ, Santin AD (2007). Advances in dendritic-cell-based therapeutic vaccines for cervical cancer. Expert Rev Anticancer Ther, 7, 1473-86. https://doi.org/10.1586/14737140.7.10.1473
  4. Calderwood SK, Theriault JR.Gong J (2005). Message in a bottle, role of the 70-kDa heat shock protein family in antitumor immunity. Eur J Immunol, 35, 2518-27. https://doi.org/10.1002/eji.200535002
  5. Ernstoff MS, Crocenzi TS, Seigne JD (2007). Developing a rational tumor vaccine therapy for renal cell carcinoma, immune yin and yang. Clin Cancer Res, 13, 733-40. https://doi.org/10.1158/1078-0432.CCR-06-2064
  6. Heiser A, Maurice M. A, Yancey D. R (2001). Human dendritic cells transfected with renal tumor RNA stimulate polyclonal T-cell responses against antigens expressed by primary and metastatic tumors. Cancer Res, 61, 3388-93.
  7. Hu YX, Li M, Jia XH, et al (2013). HPV16 CTL Epitope Peptideactivated Dendritic Cell and Natural Killer Co-culture for Therapy of Cervical Cancer in an Animal Model. Asian Pac J Cancer Prev, 14, 7335-8. https://doi.org/10.7314/APJCP.2013.14.12.7335
  8. Kim F. J, Campagna A, Khandrika L. (2008). Individualized medicine for renal cell carcinoma, establishment of primary cell line culture from surgical specimens. J Endourol, 22, 2361-6. https://doi.org/10.1089/end.2008.9703
  9. Kuwabara K, Nishishita T, Morishita M, et al (2007). Results of a phase I clinical study using dendritic cell vaccinations for thyroid cancer. Thyroid, 17, 53-8. https://doi.org/10.1089/thy.2006.0178
  10. Nadeau K, Nadler SG, Saulnier M, et al (1994). Quantitation of the interaction of the immunosuppressant deoxyspergualin and analogs with Hsc70 and Hsp90. Biochemistry, 33, 2561-7. https://doi.org/10.1021/bi00175a027
  11. Ludewig B, Ochsenbein AF, Odermatt B, et al (2000). Immunotherapy with dendritic cells directed against tumor antigens shared with normal host cells results in severe autoimmune disease. J Exp Med, 191, 795-804. https://doi.org/10.1084/jem.191.5.795
  12. Mayordomo JI, Andres R, Isla MD, et al (2007). Results of a pilot trial of immunotherapy with dendritic cells pulsed with autologous tumor lysates in patients with advanced cancer. Tumori, 93, 26-30.
  13. Nandan D, Daubenberger C, Mpimbaza G, et al (1994). A rapid, single-step purification method for immunogenic members of the hsp 70 family, validation and application. J Immunol Methods, 176, 255-63. https://doi.org/10.1016/0022-1759(94)90319-0
  14. Nadler SG, Tepper MA, Schacter B, et al (1992). Interaction of the immunosuppressant deoxyspergualin with a member of the Hsp70 family of heat shock proteins. Science, 258, 484-6. https://doi.org/10.1126/science.1411548
  15. Passalacqua R, Buti S, Tomasello G, et al (2006). Immunotherapy options in metastatic renal cell cancer, where we are and where we are going. Expert Rev Anticancer Ther, 6, 1459-72. https://doi.org/10.1586/14737140.6.10.1459
  16. Peng P, Menoret A, Srivastava PK (1997). Purification of immunogenic heat shock protein 70-peptide complexes by ADP-affinity chromatography. J Immunol Methods, 204, 13-21. https://doi.org/10.1016/S0022-1759(97)00017-3
  17. Pinzon-Charry A, Schmidt C, Lopez JA (2006). Dendritic cell immunotherapy for breast cancer. Expert Opin Biol Ther, 6, 591-604. https://doi.org/10.1517/14712598.6.6.591
  18. Pittoggi C, Martis G, Mastrangeli G, et al (2008). In vitro evidence for a new therapeutic approach in renal cell carcinoma. Int Braz J Urol, 34, 492-502. https://doi.org/10.1590/S1677-55382008000400012
  19. Ramsey S, Aitchison M (2006). Treatment for renal cancer, are we beyond the cytokine era? Nat Clin Pract Urol, 3, 478-84.
  20. Wierecky J, Muller MR, Wirths S, et al (2006). Immunologic and clinical responses after vaccinations with peptide-pulsed dendritic cells in metastatic renal cancer patients. Cancer Res, 66, 5910-8. https://doi.org/10.1158/0008-5472.CAN-05-3905
  21. Srivastava P (2002). Roles of heat-shock proteins in innate and adaptive immunity. Nat Rev Immunol, 2, 185-94. https://doi.org/10.1038/nri749
  22. Tanriverdi O (2013). Review on targeted treatment of patients with advanced-stage renal cell carcinoma, a medical oncologist's perspective. Asian Pac J Cancer Prev, 14, 609-17. https://doi.org/10.7314/APJCP.2013.14.2.609
  23. Udono H.Srivastava PK (1993). Heat shock protein 70-associated peptides elicit specific cancer immunity. J Exp Med, 178, 1391-6. https://doi.org/10.1084/jem.178.4.1391

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