Skip to main content

Thank you for visiting nature.com. You are using a browser version with limited support for CSS. To obtain the best experience, we recommend you use a more up to date browser (or turn off compatibility mode in Internet Explorer). In the meantime, to ensure continued support, we are displaying the site without styles and JavaScript.

  • Research Article
  • Published:

HSP70L1-mediated intracellular priming of dendritic cell vaccination induces more potent CTL response against cancer

Cellular & Molecular Immunology volume 15, pages 135–145 (2018)Cite this article

A Correction to this article was published on 26 November 2019

This article has been updated

Abstract

Heat-shock protein (HSP)-based immunotherapy is established on its adjuvant effects when applied via an extracellular approach to pulse and activate dendritic cells (DCs). Our previous studies indicate that DCs pulsed with recombinant fusion proteins of antigenic fragment and HSP70-like protein 1 (HSP70L1) are potent in stimulating antigen-specific Th1 responses. We herein evaluated the cytotoxic T cell (CTL) response by an intracellular approach of priming DCs with transfection of recombinant adenovirus-expressing the fusion gene of the 576–699 fragment of carcinoembryonic antigen (CEA) and HSP70L1. As compared with DCs pulsed with extracellular fusion protein, the DCs transfected with recombinant adenovirus expressing the fusion gene displayed equivalent mature phenotypes but less inflammatory appearance. However, the transfected DCs were superior to the pulsed DCs in inducing CEA-specific CTLs. Consistently, immunization of HLA-A2.1/H-2Kb transgene mice with the transfected DCs could induce more quantities of HLA-A2.1-restricted CEA-specific CTLs, protecting nude mice more significantly from human CEA-expressing colon tumor challenge when adoptively transferred. Mechanistic investigation indicated that intracellular expression of the fusion protein empowered the transfected DCs by activation of STAT1 possibly via inducing IFN-β and ERK pathways. Therefore, the more potent ability to induce anti-CEA CTL responses enables the DCs, which transfected with recombinant adenovirus expressing the fusion gene of antigenic CEA fragment and Th1 adjuvant, as an alternative promising approach for the immunotherapy of CEA-positive tumors.

This is a preview of subscription content, access via your institution

Access options

Buy this article

  • Purchase on Springer Link
  • Instant access to full article PDF
Buy now

Prices may be subject to local taxes which are calculated during checkout

Figure 1
Figure 2
Figure 3
Figure 4
Figure 5

Similar content being viewed by others

Change history

  • 26 November 2019

    "An amendment to this paper has been published and can be accessed via a link at the top of the paper."

References

  1. Doyle SM, Genest O, Wickner S . Protein rescue from aggregates by powerful molecular chaperone machines. Nat Rev Mol Cell Biol 2013; 14: 617–629.

    Article  CAS  Google Scholar 

  2. Kültz D . Molecular and evolutionary basis of the cellular stress response. Annu Rev Physiol 2005; 67: 225–257.

    Article  Google Scholar 

  3. Saibil H . Chaperone machines for protein folding, unfolding and disaggregation. Nat Rev Mol Cell Biol 2013; 14: 630–642.

    Article  CAS  Google Scholar 

  4. Kampinga HH, Craig EA . The HSP70 chaperone machinery: J proteins as drivers of functional specificity. Nat Rev Mol Cell Biol 2010; 11: 579–592.

    Article  CAS  Google Scholar 

  5. Baker-LePain JC, Sarzotti M, Nicchitta CV . Glucose-regulated protein 94/glycoprotein 96 elicits bystander activation of CD4+ T cell Th1 cytokine production in vivo. J Immunol 2004; 172: 4195–4203.

    Article  CAS  Google Scholar 

  6. Tanaka T, Okuya K, Kutomi G, Takaya A, Kajiwara T, Kanaseki T et al. Heat shock protein 90 targets a chaperoned peptide to the static early endosome for efficient cross-presentation by human dendritic cells. Cancer Sci 2015; 106: 18–24.

    Article  CAS  Google Scholar 

  7. Gong J, Zhang Y, Durfee J, Weng D, Liu C, Koido S et al. A heat shock protein 70-based vaccine with enhanced immunogenicity for clinical use. J Immunol 2010; 184: 488–496.

    Article  CAS  Google Scholar 

  8. Jung ID, Shin SJ, Lee MG, Kang TH, Han HD, Lee SJ et al. Enhancement of tumor-specific T cell-mediated immunity in dendritic cell-based vaccines by Mycobacterium tuberculosis heat shock protein X. J Immunol 2014; 193: 1233–1245.

    Article  CAS  Google Scholar 

  9. Valentinis B, Capobianco A, Esposito F, Bianchi A, Rovere-Querini P, Manfredi AA et al. Human recombinant heat shock protein 70 affects the maturation pathways of dendritic cells in vitro and has an in vivo adjuvant activity. J Leukoc Biol 2008; 84: 199–206.

    Article  CAS  Google Scholar 

  10. Binder RJ . Functions of heat shock proteins in pathways of the innate and adaptive immune system. J Immunol 2014; 193: 5765–5771.

    Article  CAS  Google Scholar 

  11. Vabulas RM, Braedel S, Hilf N, Singh-Jasuja H, Herter S, Ahmad-Nejad P et al. The endoplasmic reticulum-resident heat shock protein Gp96 activates dendritic cells via the Toll-like receptor 2/4 pathway. J Biol chem 2002; 277: 20847–20853.

    Article  CAS  Google Scholar 

  12. Saito K, Kukita K, Kutomi G, Okuya K, Asanuma H, Tabeya T et al. Heat shock protein 90 associates with Toll-like receptors 7/9 and mediates self-nucleic acid recognition in SLE. Eur J Immunol 2015; 45: 2028–2041.

    Article  CAS  Google Scholar 

  13. Gong J, Zhu B, Murshid A, Adachi H, Song B, Lee A et al. T cell activation by heat shock protein 70 vaccine requires TLR signaling and scavenger receptor expressed by endothelial cells-1. J Immunol 2009; 183: 3092–3098.

    Article  CAS  Google Scholar 

  14. Singh-Jasuja H, Toes RE, Spee P, Münz C, Hilf N, Schoenberger SP et al. Cross-presentation of glycoprotein 96-associated antigens on major histocompatibility complex class I molecules requires receptor-mediated endocytosis. J Exp Med 2000; 191: 1965–1974.

    Article  CAS  Google Scholar 

  15. Basu S, Binder RJ, Ramalingam T, Srivastava PK . CD91 is a common receptor for heat shock proteins gp96, hsp90, hsp70, and calreticulin. Immunity 2001; 14: 303–313.

    Article  CAS  Google Scholar 

  16. Xie J, Zhu H, Guo L, Ruan Y, Wang L, Sun L et al. Lectin-like oxidized low-density lipoprotein receptor-1 delivers heat shock protein 60-fused antigen into the MHC class I presentation pathway. J Immunol 2010; 185: 2306–2313.

    Article  CAS  Google Scholar 

  17. Jockheck-Clark AR, Bowers EV, Totonchy MB, Neubauer J, Pizzo SV, Nicchitta CV . Re-examination of CD91 function in GRP94 (glycoprotein 96) surface binding, uptake, and peptide cross-presentation. J Immunol 2010; 185: 6819–6830.

    Article  CAS  Google Scholar 

  18. Pawaria S, Binder RJ . CD91-dependent programming of T-helper cell responses following heat shock protein immunization. Nat Commun 2011; 2: 521.

    Article  Google Scholar 

  19. Imai T, Kato Y, Kajiwara C, Mizukami S, Ishige I, Ichiyanagi T et al. Heat shock protein 90 (HSP90) contributes to cytosolic translocation of extracellular antigen for cross-presentation by dendritic cells. Proc Natl Acad Sci USA 2011; 108: 16363–16368.

    Article  CAS  Google Scholar 

  20. Ichiyanagi T, Imai T, Kajiwara C, Mizukami S, Nakai A, Nakayama T et al. Essential role of endogenous heat shock protein 90 of dendritic cells in antigen cross-presentation. J Immunol 2010; 185: 2693–2700.

    Article  CAS  Google Scholar 

  21. Ampie L, Choy W, Lamano JB, Fakurnejad S, Bloch O, Parsa AT . Heat shock protein vaccines against glioblastoma: from bench to bedside. J Neurooncol 2015; 123: 441–448.

    Article  CAS  Google Scholar 

  22. Zhang Y, Zhang Y, Chen J, Liu Y, Luo W . Dendritic-tumor fusion cells derived heat shock protein70-peptide complex has enhanced immunogenicity. PLoS One 2015; 10: e0126075.

    Article  Google Scholar 

  23. Zhou L, Zhu T, Ye X, Yang L, Wang B, Liang X et al. Long-term protection against human papillomavirus e7-positive tumor by a single vaccination of adeno-associated virus vectors encoding a fusion protein of inactivated e7 of human papillomavirus 16/18 and heat shock protein 70. Hum Gene Ther 2010; 21: 109–119.

    Article  CAS  Google Scholar 

  24. Testori A, Richards J, Whitman E, Mann GB, Lutzky J, Camacho L et al. Phase III comparison of vitespen, an autologous tumor-derived heat shock protein gp96 peptide complex vaccine, with physician's choice of treatment for stage IV melanoma: the C-100-21 Study Group. J Clin Oncol 2008; 26: 955–962.

    Article  CAS  Google Scholar 

  25. Tischer S, Basila M, Maecker-Kolhoff B, Immenschuh S, Oelke M, Blasczyk R et al. Heat shock protein 70/peptide complexes: potent mediators for the generation of antiviral T cells particularly with regard to low precursor frequencies. J Transl Med 2011; 9: 175.

    Article  CAS  Google Scholar 

  26. Newman RG, Dee MJ, Malek TR, Podack ER, Levy RB . Heat shock protein vaccination and directed IL-2 therapy amplify tumor immunity rapidly following bone marrow transplantation in mice. Blood 2014; 123: 3045–3055.

    Article  CAS  Google Scholar 

  27. Maeda Y, Yoshimura K, Matsui H, Shindo Y, Tamesa T, Tokumitsu Y et al. Dendritic cells transfected with heat-shock protein 70 messenger RNA for patients with hepatitis C virus-related hepatocellular carcinoma: a phase 1 dose escalation clinical trial. Cancer Immunol Immunother 2015; 64: 1047–1056.

    Article  CAS  Google Scholar 

  28. Otto H, Conz C, Maier P, Wölfle T, Suzuki CK, Jenö P et al. The chaperones MPP11 and Hsp70L1 form the mammalian ribosome-associated complex. Proc Natl Acad Sci USA 2005; 102: 10064–10069.

    Article  CAS  Google Scholar 

  29. Jaiswal H, Conz C, Otto H, Wölfle T, Fitzke E, Mayer MP et al. The chaperone network connected to human ribosome-associated complex. Mol Cell Biol 2011; 31: 1160–1173.

    Article  CAS  Google Scholar 

  30. Leidig C, Bange G, Kopp J, Amlacher S, Aravind A, Wickles S et al. Structural characterization of a eukaryotic chaperone—the ribosome-associated complex. Nat Struct Mol Biol 2013; 20: 23–28.

    Article  CAS  Google Scholar 

  31. Wan T, Zhou X, chen G, An H, chen T, Zhang W et al. Novel heat shock protein Hsp70L1 activates dendritic cells and acts as a Th1 polarizing adjuvant. Blood 2004; 103: 1747–1754.

    Article  CAS  Google Scholar 

  32. Fang H, Wu Y, Huang X, Wang W, Ang B, Cao X et al. Toll-like receptor 4 (TLR4) is essential for Hsp70-like protein 1 (HSP70L1) to activate dendritic cells and induce Th1 response. J Biol chem 2011; 286: 30393–30400.

    Article  CAS  Google Scholar 

  33. Fu Q, Wu Y, Yan F, Wang N, Wang W, Cao X et al. Efficient induction of a Her2-specific anti-tumor response by dendritic cells pulsed with a Hsp70L1-Her2(341-456) fusion protein. Cell Mol Immunol 2011; 8: 424–432.

    Article  CAS  Google Scholar 

  34. Fang H, Ang B, Xu X, Huang X, Wu Y, Sun Y et al. TLR4 is essential for dendritic cell activation and anti-tumor T-cell response enhancement by DAMPs released from chemically stressed cancer cells. Cell Mol Immunol 2014; 11: 150–159.

    Article  CAS  Google Scholar 

  35. Wu Y, Wan T, Zhou X, Wang B, Yang F, Li N et al. Hsp70-like protein 1 fusion protein enhances induction of carcinoembryonic antigen-specific CD8+ CTL response by dendritic cell vaccine. Cancer Res 2005; 65: 4947–4954.

    Article  CAS  Google Scholar 

  36. Lam E, Stein S, Falck-Pedersen E . Adenovirus detection by the cGAS/STING/TBK1 DNA sensing cascade. J Virol 2014; 88: 974–981.

    Article  Google Scholar 

  37. De Simone V, Franzè E, Ronchetti G, Colantoni A, Fantini MC, Di Fusco D et al. Th17-type cytokines, IL-6 and TNF-α synergistically activate STAT3 and NF-kB to promote colorectal cancer cell growth. Oncogene 2015; 34: 3493–3503.

    Article  CAS  Google Scholar 

  38. Malanga D, De Marco C, Guerriero I, Colelli F, Rinaldo N, Scrima M et al. The Akt1/IL-6/STAT3 pathway regulates growth of lung tumor initiating cells. Oncotarget 2015; 6: 42667–42686.

    Article  Google Scholar 

  39. Deng Z, Mu J, Tseng M, Wattenberg B, Zhuang X, Egilmez NK et al. Enterobacteria-secreted particles induce production of exosome-like S1P-containing particles by intestinal epithelium to drive Th17-mediated tumorigenesis. Nat Commun 2015; 6: 6956.

    Article  CAS  Google Scholar 

  40. Wu L, Chen X, Zhao J, Martin B, Zepp JA, Ko JS et al. A novel IL-17 signaling pathway controlling keratinocyte proliferation and tumorigenesis via the TRAF4-ERK5 axis. J Exp Med 2015; 212: 1571–1587.

    Article  CAS  Google Scholar 

Download references

Acknowledgements

We thank Drs Chaofeng Han, Meng Xia and Kun Chen for technical assistance. This work was supported by grants from the National High-Tech Projects (2012AA020808 and 2007AA021003).

Author information

Author notes

  1. Lijun Song

    Present address: 3Current address: Department of Tumor, Ningxia People's Hospital, Yinchuan, China,

Authors and Affiliations

  1. National Key Laboratory of Medical Immunology, Institute of Immunology, Second Military Medical University, Shanghai, 200433, China

    Shuxun Liu, Lin Yi, Ma Ling, Jinxia Jiang, Lijun Song, Juan Liu & Xuetao Cao

  2. Department of Immunology, National Key Laboratory of Medical Molecular Biology, Chinese Academy of Medical Sciences, Beijing, 100730, China

    Xuetao Cao

Authors
  1. Shuxun Liu
    View author publications

    You can also search for this author in PubMed Google Scholar

  2. Lin Yi
    View author publications

    You can also search for this author in PubMed Google Scholar

  3. Ma Ling
    View author publications

    You can also search for this author in PubMed Google Scholar

  4. Jinxia Jiang
    View author publications

    You can also search for this author in PubMed Google Scholar

  5. Lijun Song
    View author publications

    You can also search for this author in PubMed Google Scholar

  6. Juan Liu
    View author publications

    You can also search for this author in PubMed Google Scholar

  7. Xuetao Cao
    View author publications

    You can also search for this author in PubMed Google Scholar

Corresponding authors

Correspondence to Juan Liu or Xuetao Cao.

Ethics declarations

Competing interests

The authors declare no conflict of interest.

Additional information

Supplementary Information for this article can be found on the Cellular & Molecular Immunology website

Supplementary information

Rights and permissions

Reprints and permissions

About this article

Check for updates. Verify currency and authenticity via CrossMark

Cite this article

Liu, S., Yi, L., Ling, M. et al. HSP70L1-mediated intracellular priming of dendritic cell vaccination induces more potent CTL response against cancer. Cell Mol Immunol 15, 135–145 (2018). https://doi.org/10.1038/cmi.2016.33

Download citation

  • Received:

  • Revised:

  • Accepted:

  • Published:

  • Issue Date:

  • DOI: https://doi.org/10.1038/cmi.2016.33

Keywords

This article is cited by

Search

Advanced search

Quick links