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Killed but metabolically active microbes: a new vaccine paradigm for eliciting effector T-cell responses and protective immunity

Nature Medicine volume 11, pages 853–860 (2005)Cite this article

Abstract

We developed a new class of vaccines, based on killed but metabolically active (KBMA) bacteria, that simultaneously takes advantage of the potency of live vaccines and the safety of killed vaccines. We removed genes required for nucleotide excision repair (uvrAB), rendering microbial-based vaccines exquisitely sensitive to photochemical inactivation with psoralen and long-wavelength ultraviolet light. Colony formation of the nucleotide excision repair mutants was blocked by infrequent, randomly distributed psoralen crosslinks, but the bacterial population was able to express its genes, synthesize and secrete proteins. Using the intracellular pathogen Listeria monocytogenes as a model platform, recombinant psoralen-inactivated Lm ΔuvrAB vaccines induced potent CD4+ and CD8+ T-cell responses and protected mice against virus challenge in an infectious disease model and provided therapeutic benefit in a mouse cancer model. Microbial KBMA vaccines used either as a recombinant vaccine platform or as a modified form of the pathogen itself may have broad use for the treatment of infectious disease and cancer.

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Figure 1: L. monocytogenes nucleotide excision repair mutants are exquisitely sensitive to photochemical inactivation and are metabolically active.
Figure 2: Photochemically inactivated L. monocytogenes nucleotide excision repair mutant vaccines induce class I reponses.
Figure 3: S-59/UVA Lm ΔactAΔuvrAB-OVA elicit functional CD8+ T cells in vaccinated mice and protect against a subsequent viral challenge.
Figure 4: Nucleotide excision repair mutation confers therapeutic efficacy of S-59/UVA Lm ΔactAΔuvrAB-AH1-A5 vaccines in tumor-bearing animals.
Figure 5: S-59/UVA Lm ΔactA/ΔuvrAB vaccines protect immunized mice against wild-type L. monocytogenes challenge.

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Acknowledgements

We wish to thank D. Pardoll, J. Skoble, P. Lauer and A. North for their discussions, suggestions and critical review of this manuscript, and J. Cox for his suggestion of the KBMA acronym.

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Author notes

  1. D G Brockstedt, K S Bahjat and M A Giedlin: These authors contributed equally to this work.

Authors and Affiliations

  1. Cerus Corporation, 2411 Stanwell Drive, Concord, 94520, California, USA

    D G Brockstedt, K S Bahjat, M A Giedlin, W Liu, M Leong, W Luckett, Y Gao, D Kapadia, G Castro, J Y H Lim, A Sampson-Johannes, A Stassinopoulos, J E Hearst, D N Cook & T W Dubensky Jr.

  2. Department of Molecular and Cell Biology, Oregon Health Sciences University, 3710 SW US Veterans Hospital Road, Portland, 97201, Oregon, USA

    P Schnupf, A A Herskovits, H G Archie Bouwer & D A Portnoy

  3. School of Public Health, University of California, 508 Barker Hall, #3202, Berkeley, 94720-3202, California, USA

    D A Portnoy

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Correspondence to T W Dubensky Jr..

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Competing interests

D.G. Brockstedt, K.S. Bahjat, M.A. Giedlin, W. Liu, M. Leong, W. Luckett, Y. Gao, G. Castro, J.Y.H. Lim, A. Sampson-Johannes, A. Stassinopoulos, J.E. Hearst, D.N. Cook and T.W. Dubensky, Jr.are employees of Cerus Corporation, which owns intellectual property covering the compositions and methods described in this manuscript. In addition, Cerus employees hold stock and/or stock options in the company. D.A. Portnoy is is a paid consultant to the company and holds stock options. The remaining authors have no known financial interest in Cerus.

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Veterans Affairs Medical Center, Department of Molecular Microbiology and Immunology, Oregon Health Sciences University, 3710 SW US Veterans Hospital Road, Portland, Oregon 97201, USA.

Supplementary information

Supplementary Fig. 1

Psoralen inactivated Lm actA/uvrAB continue to express and to secrete proteins after photochemical inactivation. (PDF 1234 kb)

Supplementary Fig. 2

Psoralen inactivated Lm actA/uvrAB -AH1-A5 induce cytotoxic T cells in vivo. (PDF 1446 kb)

Supplementary Fig. 3

KBMA B.anthracis uvrAB (Sterne)are exquisitely sensitive to photochemical inactivation. (PDF 1037 kb)

Supplementary Methods (PDF 21 kb)

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Brockstedt, D., Bahjat, K., Giedlin, M. et al. Killed but metabolically active microbes: a new vaccine paradigm for eliciting effector T-cell responses and protective immunity. Nat Med 11, 853–860 (2005). https://doi.org/10.1038/nm1276

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