HIV-1 reverse transcriptase artificially targeted for proteasomal degradation induces a mixed Th1/Th2-type immune response

Starodubova, Elizaveta; Boberg, Andreas; Литвина, М М; Morozov, Alexey; Петракова, Н. В.; Timofeev, A.; Latyshev, Oleg; Tunitskaya, V. L.; Wahrén, Britta; Isaguliants, Maria; Карпов, В.Л.

doi:10.1016/j.vaccine.2008.03.070

Cited by 20 publications

(30 citation statements)

References 32 publications

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“…Potent immunogenic performance of all three HIV enzymes is a prerequisite of the efficacy of such immunotherapy. We and others performed series of studies aimed to improve the immunogenicity of RT, a key enzyme determining HIV-1 resistance to antiretroviral therapy, but with a limited success [12, 13, 16–18]. Lately, we found that cells expressing HIV-1 RT produce reactive oxygen species (ROS) and express high levels of phase II detoxifying enzymes that interfere with the immune response against this enzyme [19, 20].…”

Section: Introductionmentioning

confidence: 99%

Fusion to Flaviviral Leader Peptide Targets HIV-1 Reverse Transcriptase for Secretion and Reduces Its Enzymatic Activity and Ability to Induce Oxidative Stress but Has No Major Effects on Its Immunogenic Performance in DNA-Immunized Mice

Latanova

Petkov

Kuzmenko

et al. 2017

Journal of Immunology Research

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Reverse transcriptase (RT) is a key enzyme in viral replication and susceptibility to ART and a crucial target of immunotherapy against drug-resistant HIV-1. RT induces oxidative stress which undermines the attempts to make it immunogenic. We hypothesized that artificial secretion may reduce the stress and make RT more immunogenic. Inactivated multidrug-resistant RT (RT1.14opt-in) was N-terminally fused to the signal providing secretion of NS1 protein of TBEV (Ld) generating optimized inactivated Ld-carrying enzyme RT1.14oil. Promotion of secretion prohibited proteasomal degradation increasing the half-life and content of RT1.14oil in cells and cell culture medium, drastically reduced the residual polymerase activity, and downmodulated oxidative stress. BALB/c mice were DNA-immunized with RT1.14opt-in or parental RT1.14oil by intradermal injections with electroporation. Fluorospot and ELISA tests revealed that RT1.14opt-in and RT1.14oil induced IFN-γ/IL-2, RT1.14opt-in induced granzyme B, and RT1.14oil induced perforin production. Perforin secretion correlated with coproduction of IFN-γ and IL-2 (R = 0,97). Both DNA immunogens induced strong anti-RT antibody response. Ld peptide was not immunogenic. Thus, Ld-driven secretion inferred little change to RT performance in DNA immunization. Positive outcome was the abrogation of polymerase activity increasing safety of RT-based DNA vaccines. Identification of the molecular determinants of low cellular immunogenicity of RT requires further studies.

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Section: Introductionmentioning

confidence: 99%

Fusion to Flaviviral Leader Peptide Targets HIV-1 Reverse Transcriptase for Secretion and Reduces Its Enzymatic Activity and Ability to Induce Oxidative Stress but Has No Major Effects on Its Immunogenic Performance in DNA-Immunized Mice

Latanova

Petkov

Kuzmenko

et al. 2017

Journal of Immunology Research

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“…The use of protein degradation signals, which direct proteins to the proteasome via an ubiquitin–independent mechanism, circumvents the effects of a multitude of factors and thus provides a more straight–forward way of proteasome targeting. Fusion of HIV–1 reverse transcriptase with the minimal proteasome–targeting signals of ODC represented by two short amino acid sequences at the ODC C–terminus led to an accelerated degradation and an increased immunogenicity of the chimeric gene in mice as compared to the original gene [40]. This modification was also successful when applied to the weakly immunogenic reverse transcriptase of drug–resistant HIV–1 [41] helping to enhance both cellular and antibody immune responses against the mutant enzyme form [42].…”

Section: Ubiquitin-independent Mechanismmentioning

confidence: 99%

Regulation of immunogen processing: signal sequences and their application for the new generation of DNA-vaccines

2010

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Immunization with naked genes (DNA–immunization) is a perspective modern approach to prophylactic as well as therapeutic vaccination against pathogens, as well as cancer and allergy. A panel of DNA immunogens has been developed, some are already in the clinical trials. However, the immunogenicity of DNA vaccines, specifically of those applied to humans, needs a considerable improvement. There are several approaches to increase DNA vaccine immunogenicity. One approach implies the modifications of the encoded immunogen that change its processing and presentation, and thus the overall pattern of anti–immunogen response. For this, eukaryotic expression vectors are constructed that encode the chimeric proteins composed of the immunogen and specialized targeting or signal sequences. The review describes a number of signals that if fused to immunogen, target it into the predefined subcellular compartments. The review gives examples of their application for DNA–immunization.

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“…Chimeric fusion proteins fused to the N terminus of ODC promote the proteasome-dependent degradation of ODC and their interacting cellular target proteins (Matsuzawa et al, 2005). Moreover, it was reported that mice immunized with DNA encoding HIV-1 reverse transcriptase (RT) fused to ODC or the ODC signal domain showed marked enhancement of RT degradation and the immune response (Starodubova et al, 2008).…”

Section: Introductionmentioning

confidence: 99%

“…In addition, mouse ODC fusion proteins were tested by transfection of human embryonic kidney cells (HEK293), and the effect of ODC activity on proteasomal degradation was confirmed by Western blot and cycloheximide chase experiments ( Johnson et al, 1992). There is also a report that antigen fusion with the signal domain of ODC induces a stronger immune response than fusion with full-length ODC (Starodubova et al, 2008). The C-terminal region of ODC (amino acids 423-461), which contains PEST domains, is essential for its degradation (Murakami et al, 2000).…”

mentioning

confidence: 99%

Fusion of the Human Cytomegalovirus pp65 Antigen with Both Ubiquitin and Ornithine Decarboxylase Additively Enhances Antigen Presentation to CD8⁺T Cells in Human Dendritic Cells

et al. 2010

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Antigenic molecules are modified for targeting to the proteasome by ubiquitin (Ub) or by a Ub-independent system such as ornithine decarboxylase (ODC) to be presented by MHC class I molecules. In this study, we compared the immunogenicity of human cytomegalovirus pp65 antigen fused with Ub and/or ODC, using RNA electroporation of human dendritic cells. Among the C-terminal mutants of Ub (G76, A76, and V76), Ub(G) showed the best ability to enhance the degradation of a target protein and stimulate T cells. The pp65 antigens fused with either Ub(G) or ODC enhanced the stimulation to CD8(+) T cells, and the effects of Ub(G) and ODC were similar. Furthermore, the fusion of both Ub and ODC additively increased immunogenicity compared with the single-fusion proteins. The fusion of Ub(G) and ODC enhanced primarily the stimulation of CD8(+) rather than CD4(+) T cells and more efficiently induced pp65-specific T cells in vitro. These additive effects of Ub and ODC in antigen processing may provide improved strategies to stimulate CD8(+) T cells for the development of immunotherapies against the variety of viral diseases and cancers.

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HIV-1 reverse transcriptase artificially targeted for proteasomal degradation induces a mixed Th1/Th2-type immune response

Cited by 20 publications

References 32 publications

Fusion to Flaviviral Leader Peptide Targets HIV-1 Reverse Transcriptase for Secretion and Reduces Its Enzymatic Activity and Ability to Induce Oxidative Stress but Has No Major Effects on Its Immunogenic Performance in DNA-Immunized Mice

Fusion to Flaviviral Leader Peptide Targets HIV-1 Reverse Transcriptase for Secretion and Reduces Its Enzymatic Activity and Ability to Induce Oxidative Stress but Has No Major Effects on Its Immunogenic Performance in DNA-Immunized Mice

Regulation of immunogen processing: signal sequences and their application for the new generation of DNA-vaccines

Fusion of the Human Cytomegalovirus pp65 Antigen with Both Ubiquitin and Ornithine Decarboxylase Additively Enhances Antigen Presentation to CD8⁺T Cells in Human Dendritic Cells

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HIV-1 reverse transcriptase artificially targeted for proteasomal degradation induces a mixed Th1/Th2-type immune response

Cited by 20 publications

References 32 publications

Fusion to Flaviviral Leader Peptide Targets HIV-1 Reverse Transcriptase for Secretion and Reduces Its Enzymatic Activity and Ability to Induce Oxidative Stress but Has No Major Effects on Its Immunogenic Performance in DNA-Immunized Mice

Fusion to Flaviviral Leader Peptide Targets HIV-1 Reverse Transcriptase for Secretion and Reduces Its Enzymatic Activity and Ability to Induce Oxidative Stress but Has No Major Effects on Its Immunogenic Performance in DNA-Immunized Mice

Regulation of immunogen processing: signal sequences and their application for the new generation of DNA-vaccines

Fusion of the Human Cytomegalovirus pp65 Antigen with Both Ubiquitin and Ornithine Decarboxylase Additively Enhances Antigen Presentation to CD8+T Cells in Human Dendritic Cells

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Fusion of the Human Cytomegalovirus pp65 Antigen with Both Ubiquitin and Ornithine Decarboxylase Additively Enhances Antigen Presentation to CD8⁺T Cells in Human Dendritic Cells