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Apoptosis-mediated enhancement of DNA-raised immune responses by mutant caspases

Nature Biotechnology volume 19, pages 543–547 (2001)Cite this article

Abstract

Apoptotic bodies can be used to target delivery of DNA-expressed immunogens into professional antigen-presenting cells (APCs). Here we show that antigen-laden apoptotic bodies created by vectors co-expressing influenza virus hemagglutinin (HA) or nucleoprotein (NP) genes and mutant caspase genes markedly increased T-cell responses. Both CD8 and CD4 T-cell responses were affected. The adjuvant activity was restricted to partially inactivated caspases that allowed immunogen expression before the generation of apoptotic bodies. Active-site mutants of murine caspase 2 and an autocatalytic chimera of murine caspase 2 prodomain and human caspase 3 induced apoptosis that did not interfere with immunogen expression. The adjuvant activity also enhanced B-cell responses, but to a lesser extent than T-cell responses. The large increases in T-cell responses represent one of the strongest effects to date of a DNA adjuvant on cellular immunity.

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Figure 1: Schematic of caspase clones and dual expression vector.
Figure 2: Effect of various caspase clones on apoptosis induction and co-delivered reporter gene expression in transiently transfected cells.
Figure 3: Effect of co-expressed caspase genes on immune responses to influenza hemagglutinin.
Figure 4: Effect of co-expressed caspase genes on immune responses to influenza nucleoprotein.
Figure 5: Dose/response study for caspase effects on immune responses.
Figure 6: Adjuvant activity of the mutant caspase 3 for an influenza virus challenge.

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Acknowledgements

This work was supported by US Public Health Service research grant R01AI34946 and base grant support to Yerkes Regional Primate Research Center, RR00165. We would like to thank H. Drake-Perrow for expert administrative assistance.

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  1. Shin Sasaki

    Present address: Department of Bioregulation, Leprosy Research Center, National Institute of Infectious Diseases, 4-2*1 Aoba-cho, Tokyo, 182-0002, Japan

Authors and Affiliations

  1. Division of Microbiology and Immunology, Yerkes Regional Primate Research Center of Emory University, Atlanta, 30329, GA

    Shin Sasaki, Rama R. Amara, Alp E. Oran, James M. Smith & Harriet L. Robinson

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Correspondence to Harriet L. Robinson.

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Sasaki, S., Amara, R., Oran, A. et al. Apoptosis-mediated enhancement of DNA-raised immune responses by mutant caspases. Nat Biotechnol 19, 543–547 (2001). https://doi.org/10.1038/89289

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