Abstract
Heat shock proteins (Hsp) of the Hsp70/90 families facilitate cellular immune responses to antigenic peptides or proteins bound to them and have therefore been used as vaccine vehicles. We developed an expression system in which chimeric proteins with an Hsp-capturing, viral J domain fused to diverse antigen-encoding sequences form stable complexes with eukaryotic (Hsp70, Hsp73) or bacterial (DnaK) stress proteins and accumulate to high steady-state levels. J domains from different species (viruses/SV40, bacteria/Chlamydia trachomatis or plants/Arabidopsis thaliana) efficiently capture murine or human stress proteins in this system, thus making different J domains available for vaccine production. A novel expression and purification method was developed to produce native Hsp/antigen complexes in transfectants. These purified Hsp/antigen complexes efficiently elicited antigen-specific CD8 T cell responses in mice when delivered as vaccines without adjuvants. In situ complex formation of antigen with Hsp was critical for CD8 T cell priming. Because the described expression system supports the flexible design of multivalent vaccines, it is an attractive strategy to elicit CD8 T cell responses either to recombinant proteins or to selected antigenic domains of these molecules.
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Abbreviations
- Ad:
-
adenovirus
- APC:
-
antigen-presenting cells
- DC:
-
dendritic cell
- eGFP (or G):
-
enhanced green fluorescent protein
- FCM:
-
flow cytometry
- Flu (or F):
-
influenza A
- HBsAg (or S):
-
hepatitis B surface antigen
- HBV:
-
hepatitis B virus
- Hsp:
-
heat shock protein
- LPS:
-
lipopolysaccharide
- ODN:
-
oligodeoxynucleotide
- OVA:
-
ovalbumin
- Pol (or P):
-
hepatitis B polymerase
- SV40:
-
simian virus 40
- T-Ag (or T):
-
large tumor antigen of SV40
- wt:
-
wild type
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Acknowledgments
We thank K. Ölberger and C. Heilig for outstanding technical assistance. We thank Dr. F. Lemonnier (Institut Pasteur, Paris, France) for the HLA-A*0201 transgenic mice and A. Schreiber (Dept. of Virology, University of Ulm, Germany) for the bacterial expressed, recombinant eGFP-protein. This work was supported by a grant from the Deutsche Forschungsgemeinschaft (SCHI-505/2-4) to R.S.
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Wieland, A., Denzel, M., Schmidt, E. et al. Recombinant complexes of antigen with stress proteins are potent CD8 T-cell-stimulating immunogens. J Mol Med 86, 1067–1079 (2008). https://doi.org/10.1007/s00109-008-0371-x
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DOI: https://doi.org/10.1007/s00109-008-0371-x