Limits...
Recombinant rabies virus particles presenting botulinum neurotoxin antigens elicit a protective humoral response in vivo.

Hudacek AW, Al-Saleem FH, Willet M, Eisemann T, Mattis JA, Simpson LL, Schnell MJ - Mol Ther Methods Clin Dev (2014)

Bottom Line: Our previous research has shown that recombinant rabies virus-based particles can effectively present heterologous antigens.Administration of a combination of these vectors also demonstrated antibody responses against all three serotypes based on enzyme-linked immunosorbent assay (ELISA) measurements, with minimal decay within the study timeline.Complete protection was achieved against toxin challenge from the serotypes /A and /B and partial protection for /E, indicating that a multivalent approach is feasible.

View Article: PubMed Central - PubMed

Affiliation: Molecular Targeting Technologies, Inc. , West Chester, Pennsylvania, USA ; Department of Microbiology and Immunology, Sidney Kimmel Medical College, Thomas Jefferson University , Philadelphia, Pennsylvania, USA.

ABSTRACT
Botulinum neurotoxins are one of the most potent toxins found in nature, with broad medical applications from cosmetics to the treatment of various neuropathies. Additionally, these toxins are classified as Category A-Tier 1 agents, with human lethal doses calculated at as little as 90 ng depending upon the route of administration. Of the eight distinct botulinum neurotoxin serotypes, the most common causes of human illness are from serotypes /A, /B, and /E. Protection can be achieved by eliciting antibody responses against the receptor-binding domain of the neurotoxin. Our previous research has shown that recombinant rabies virus-based particles can effectively present heterologous antigens. Here, we describe a novel strategy using recombinant rabies virus particles that elicits a durable humoral immune response against the botulinum neurotoxin receptor binding domains from serotypes /A, /B, and /E. Following intramuscular administration of β-propiolactone-inactivated rabies virus particles, mice elicited specific immune responses against the cognate antigen. Administration of a combination of these vectors also demonstrated antibody responses against all three serotypes based on enzyme-linked immunosorbent assay (ELISA) measurements, with minimal decay within the study timeline. Complete protection was achieved against toxin challenge from the serotypes /A and /B and partial protection for /E, indicating that a multivalent approach is feasible.

No MeSH data available.


Related in: MedlinePlus

Multistep growth kinetics of recombinant viruses on Vero cells. Vero cells were infected at a multiplicity of infection of 0.01 for 2 hours with BNSP-333 (black), BNSP-333-coHC50/A (red), BNSP-333-coHC50/B (green), or BNSP-333-HC50/E (blue). Samples were collected at the time points indicated and viral titers were determined in duplicate. FFU, fluorescent focus units.
© Copyright Policy - open-access
Related In: Results  -  Collection

License
getmorefigures.php?uid=PMC4362357&req=5

fig4: Multistep growth kinetics of recombinant viruses on Vero cells. Vero cells were infected at a multiplicity of infection of 0.01 for 2 hours with BNSP-333 (black), BNSP-333-coHC50/A (red), BNSP-333-coHC50/B (green), or BNSP-333-HC50/E (blue). Samples were collected at the time points indicated and viral titers were determined in duplicate. FFU, fluorescent focus units.

Mentions: As a potential vaccine product, the growth of recombinant viruses expressing the BoNT HC50 domains was analyzed on Vero cells, a continuous cell line approved for vaccine production.33,34 We rerecovered the following viruses on Vero cells: BNSP-333-coHC50/A, BNSP-333-coHC50/B and BNSP-333-HC50/E before analyzing their growth on Vero cells. When inoculated at a low MOI (0.01), we observed that there was little difference between the growth profiles of the parental BNSP-333 vector and the BNSP-333-coHC50/B or BNSP-333-HC50/E viruses (Figure 4). However, we detected an ~1-log difference in peak titers when comparing the parental virus to BNSP-333-coHC50/A (Figure 4, compare black circle and red diamond curves). This growth reduction was confirmed with single-step growth kinetics, in which BNSP-333-coHC50/A virus again grew to ~1-log lower peak titer compared with the parental virus (data not shown). This contrasts with the analysis of the wild type HC50/A expressing virus, which replicated with similar kinetics as the parental virus (Figure 4, compare grey diamond (wild-type HC50/A) to black circle curves (parental)). These results indicate that codon optimization of HC50/A, while allowing for increasing cell-surface expression (Figure 2a) has a detrimental impact on virus growth kinetics. However, the titer of BNSP-333-coHC50/A was still sufficient to produce the vector in the necessary amounts for vaccinations and other studies.


Recombinant rabies virus particles presenting botulinum neurotoxin antigens elicit a protective humoral response in vivo.

Hudacek AW, Al-Saleem FH, Willet M, Eisemann T, Mattis JA, Simpson LL, Schnell MJ - Mol Ther Methods Clin Dev (2014)

Multistep growth kinetics of recombinant viruses on Vero cells. Vero cells were infected at a multiplicity of infection of 0.01 for 2 hours with BNSP-333 (black), BNSP-333-coHC50/A (red), BNSP-333-coHC50/B (green), or BNSP-333-HC50/E (blue). Samples were collected at the time points indicated and viral titers were determined in duplicate. FFU, fluorescent focus units.
© Copyright Policy - open-access
Related In: Results  -  Collection

License
Show All Figures
getmorefigures.php?uid=PMC4362357&req=5

fig4: Multistep growth kinetics of recombinant viruses on Vero cells. Vero cells were infected at a multiplicity of infection of 0.01 for 2 hours with BNSP-333 (black), BNSP-333-coHC50/A (red), BNSP-333-coHC50/B (green), or BNSP-333-HC50/E (blue). Samples were collected at the time points indicated and viral titers were determined in duplicate. FFU, fluorescent focus units.
Mentions: As a potential vaccine product, the growth of recombinant viruses expressing the BoNT HC50 domains was analyzed on Vero cells, a continuous cell line approved for vaccine production.33,34 We rerecovered the following viruses on Vero cells: BNSP-333-coHC50/A, BNSP-333-coHC50/B and BNSP-333-HC50/E before analyzing their growth on Vero cells. When inoculated at a low MOI (0.01), we observed that there was little difference between the growth profiles of the parental BNSP-333 vector and the BNSP-333-coHC50/B or BNSP-333-HC50/E viruses (Figure 4). However, we detected an ~1-log difference in peak titers when comparing the parental virus to BNSP-333-coHC50/A (Figure 4, compare black circle and red diamond curves). This growth reduction was confirmed with single-step growth kinetics, in which BNSP-333-coHC50/A virus again grew to ~1-log lower peak titer compared with the parental virus (data not shown). This contrasts with the analysis of the wild type HC50/A expressing virus, which replicated with similar kinetics as the parental virus (Figure 4, compare grey diamond (wild-type HC50/A) to black circle curves (parental)). These results indicate that codon optimization of HC50/A, while allowing for increasing cell-surface expression (Figure 2a) has a detrimental impact on virus growth kinetics. However, the titer of BNSP-333-coHC50/A was still sufficient to produce the vector in the necessary amounts for vaccinations and other studies.

Bottom Line: Our previous research has shown that recombinant rabies virus-based particles can effectively present heterologous antigens.Administration of a combination of these vectors also demonstrated antibody responses against all three serotypes based on enzyme-linked immunosorbent assay (ELISA) measurements, with minimal decay within the study timeline.Complete protection was achieved against toxin challenge from the serotypes /A and /B and partial protection for /E, indicating that a multivalent approach is feasible.

View Article: PubMed Central - PubMed

Affiliation: Molecular Targeting Technologies, Inc. , West Chester, Pennsylvania, USA ; Department of Microbiology and Immunology, Sidney Kimmel Medical College, Thomas Jefferson University , Philadelphia, Pennsylvania, USA.

ABSTRACT
Botulinum neurotoxins are one of the most potent toxins found in nature, with broad medical applications from cosmetics to the treatment of various neuropathies. Additionally, these toxins are classified as Category A-Tier 1 agents, with human lethal doses calculated at as little as 90 ng depending upon the route of administration. Of the eight distinct botulinum neurotoxin serotypes, the most common causes of human illness are from serotypes /A, /B, and /E. Protection can be achieved by eliciting antibody responses against the receptor-binding domain of the neurotoxin. Our previous research has shown that recombinant rabies virus-based particles can effectively present heterologous antigens. Here, we describe a novel strategy using recombinant rabies virus particles that elicits a durable humoral immune response against the botulinum neurotoxin receptor binding domains from serotypes /A, /B, and /E. Following intramuscular administration of β-propiolactone-inactivated rabies virus particles, mice elicited specific immune responses against the cognate antigen. Administration of a combination of these vectors also demonstrated antibody responses against all three serotypes based on enzyme-linked immunosorbent assay (ELISA) measurements, with minimal decay within the study timeline. Complete protection was achieved against toxin challenge from the serotypes /A and /B and partial protection for /E, indicating that a multivalent approach is feasible.

No MeSH data available.


Related in: MedlinePlus