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Merck
  • Plant virus particles carrying tumour antigen activate TLR7 and Induce high levels of protective antibody.

Plant virus particles carrying tumour antigen activate TLR7 and Induce high levels of protective antibody.

PloS one (2015-02-19)
Jantipa Jobsri, Alex Allen, Deepa Rajagopal, Michael Shipton, Kostya Kanyuka, George P Lomonossoff, Christian Ottensmeier, Sandra S Diebold, Freda K Stevenson, Natalia Savelyeva
초록

Induction of potent antibody is the goal of many vaccines targeted against infections or cancer. Modern vaccine designs that use virus-like particles (VLP) have shown efficacy for prophylactic vaccination against virus-associated cancer in the clinic. Here we used plant viral particles (PVP), which are structurally analogous to VLP, coupled to a weak idiotypic (Id) tumour antigen, as a conjugate vaccine to induce antibody against a murine B-cell malignancy. The Id-PVP vaccine incorporates a natural adjuvant, the viral ssRNA, which acts via TLR7. It induced potent protective anti-Id antibody responses in an in vivo mouse model, superior to the "gold standard" Id vaccine, with prevalence of the IgG2a isotype. Combination with alum further increased antibody levels and maintained the IgG2a bias. Engagement of TLR7 in vivo was followed by secretion of IFN-α by plasmacytoid dendritic cells and by activation of splenic CD11chi conventional dendritic cells. The latter was apparent from up-regulation of co-stimulatory molecules and from secretion of a wide range of inflammatory cytokines and chemokines including the Th1-governing cytokine IL-12, in keeping with the IgG2a antibody isotype distribution. PVP conjugates are a novel cancer vaccine design, offering an attractive molecular form, similar to VLP, and providing T-cell help. In contrast to VLP, they also incorporate a safe "in-built" ssRNA adjuvant.

MATERIALS
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Supelco
Aphidicolin, analytical standard
Sigma-Aldrich
Brefeldin A, from Penicillium brefeldianum, Ready Made Solution, 10 mg/mL in DMSO
Sigma-Aldrich
Glutaric dialdehyde solution, 50 wt. % in H2O, FCC
Sigma-Aldrich
Glutaraldehyde solution, 50 wt. % in H2O
Sigma-Aldrich
Brefeldin A, ≥99% (HPLC and TLC), BioXtra, for molecular biology
Sigma-Aldrich
Aphidicolin from Nigrospora sphaerica, ≥98% (HPLC), powder
Sigma-Aldrich
Aluminum potassium sulfate dodecahydrate, BioXtra, ≥98.0%
Sigma-Aldrich
Glutaraldehyde solution, Grade I, 70% in H2O, specially purified for use as an electron microscopy fixative or other sophisticated use
Sigma-Aldrich
Brefeldin A, from Penicillium brefeldianum, ≥99% (HPLC and TLC)
Sigma-Aldrich
Glutaraldehyde solution, Grade I, 25% in H2O, specially purified for use as an electron microscopy fixative
Sigma-Aldrich
Glutaraldehyde solution, Grade I, 50% in H2O, specially purified for use as an electron microscopy fixative or other sophisticated use
Sigma-Aldrich
Glutaraldehyde solution, Grade II, 25% in H2O
Sigma-Aldrich
Glutaraldehyde solution, Grade I, 8% in H2O, specially purified for use as an electron microscopy fixative or other sophisticated use
Sigma-Aldrich
L-Methionine sulfoximine, suitable for cell culture
Sigma-Aldrich
Glutaraldehyde solution, 50% in H2O, suitable for photographic applications
Sigma-Aldrich
Aluminum potassium sulfate dodecahydrate, BioReagent, suitable for insect cell culture
Sigma-Aldrich
Aluminum potassium sulfate dodecahydrate, ACS reagent, ≥98%
Supelco
L-Methionine sulfoximine, certified reference material, TraceCERT®, Manufactured by: Sigma-Aldrich Production GmbH, Switzerland
Sigma-Aldrich
Glutaraldehyde solution, technical, ~50% in H2O (5.6 M)
Sigma-Aldrich
Aluminum potassium sulfate dodecahydrate, puriss. p.a., ACS reagent, reag. Ph. Eur., ≥99.5%
Sigma-Aldrich
Aluminum hydroxide, reagent grade