In vivo protein expression and immune responses generated by DNA vaccines expressing mycobacterial antigens fused with a reporter protein

Quinn, Anita; Jiang, Weiwen; Velaz-Faircloth, Maria; Cobb, Alison J; Henry, Stanley C.; Frothingham, Richard

doi:10.1016/s0264-410x(02)00253-0

Cited by 7 publications

(4 citation statements)

References 21 publications

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“…The reason for this apparent decrease in IgG2a is unclear and may indicate that the addition of eGFP alters the T‐helper immune microenvironment. One study (51), for example, found that the addition of eGFP to Mycobacterium, bovis antigen 85a could influence the IgG isotype profile. In their model, however, they observed a decrease in IgG1 levels (instead of IgG2a) when eGFP was added to the C terminus of their native protein.…”

Section: Discussionmentioning

confidence: 99%

Generation and characterization of a trackable plant-made influenza H5 virus-like particle (VLP) containing enhanced green fluorescent protein (eGFP)

Young¹,

Arthus-Cartier²,

Yam³

et al. 2015

The FASEB Journal

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Medicago, Inc. has developed an efficient virus-like particle (VLP) vaccine production platform using the Nicotiana benthamiana expression system, and currently has influenza-based products targeting seasonal/pandemic hemagglutinin (HA) proteins in advanced clinical trials. We wished to generate a trackable HA-based VLP that would allow us to study both particle assembly in plants and VLP interactions within the mammalian immune system. To this end, a fusion protein was designed, composed of H5 (from influenza A/Indonesia/05/2005 [H5N1]) with enhanced green fluorescent protein (eGFP). Expression of H5-eGFP in N. benthamiana produced brightly fluorescent ∼160 nm particles resembling H5-VLPs. H5-eGFP-VLPs elicited anti-H5 serologic responses in mice comparable to those elicited by H5-VLPs in almost all assays tested (hemagglutination inhibition/IgG(total)/IgG1/IgG2b/IgG2a:IgG1 ratio), as well as a superior anti-GFP IgG response (mean optical density = 2.52 ± 0.16 sem) to that elicited by soluble GFP (mean optical density = 0.12 ± 0.06 sem). Confocal imaging of N. benthamiana cells expressing H5-eGFP displayed large fluorescent accumulations at the cell periphery, and draining lymph nodes from mice given H5-eGFP-VLPs via footpad injection demonstrated bright fluorescence shortly after administration (10 min), providing proof of concept that the H5-eGFP-protein/VLPs could be used to monitor both VLP assembly and immune trafficking. Given these findings, this novel fluorescent reagent will be a powerful tool to gain further fundamental insight into the biology of influenza VLP vaccines.

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Section: Discussionmentioning

confidence: 99%

Generation and characterization of a trackable plant-made influenza H5 virus-like particle (VLP) containing enhanced green fluorescent protein (eGFP)

Young¹,

Arthus-Cartier²,

Yam³

et al. 2015

The FASEB Journal

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“…Because CD8 cells respond to intracellular Ags, it has been widely assumed that the induction of CD8-mediated immunity requires intracellular expression of Ag within professional APCs. DNA vaccines and recombinant viral particles have been used to achieve this in vivo (11,12). However, it is clear that dendritic cells (DCs), 3 macrophages, and activated B cells (13)(14)(15) are capable of presenting exogenously acquired Ag in the context of MHC class I to CD8 cells.…”

mentioning

confidence: 99%

Combined Triggering of Dendritic Cell Receptors Results in Synergistic Activation and Potent Cytotoxic Immunity

Wells

Cowled

Farzaneh

et al. 2008

The Journal of Immunology

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Elimination of malignant cells and intracellular infections involves collaboration between CTLs and Th1 inflammation. Dendritic cells drive this response via costimulation and cytokines. We have defined key signals required for the exponential expansion of specific CD8+ T cells in vivo in mice. Immunization with two or more TLR agonists, anti-CD40, IFN-γ, and surfactant were sufficient to drive unprecedented levels of CD8 response to peptide or protein Ag and highly polarized Th1 CD4 responses. CD40 signaling was required for CD8 expansion but could be provided by a concomitant CD4 Th response in place of anti-CD40. Triggering of these pathways activated migration and activation of myeloid and plasmacytoid dendritic cells and secretion of IL-12. Cross-presentation can thus be exploited to induce potent cytotoxic responses and long-term memory to peptide/protein Ags. When combined with a tumor-associated peptide from tyrosinase-related protein 2, our combined adjuvant approach effectively halted tumor growth in an in vivo melanoma model and was more effective than anti-CD40 and a single TLR agonist. Antitumor immunity was associated with long-lived effector memory CD8 cells specific for the naturally processed and presented tumor Ag, and tumor protection was partially but not entirely dependent on CD8 T cells. This flexible strategy is more effective than existing adjuvants and provides a technological platform for rapid vaccine development.

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“…A Th1 type response has also been reported following i.m. immunization with a plasmid encoding mycobacterial antigens (Quinn et al, 2002) or influenza virus (Johnson et al, 2000). The strain of mice used in immunization studies may affect the Th response observed as it has been reported that BALB/c mice tend to generate a Th1 response and C57BL/6 and CBA/Ca mice tend to generate Th1 and Th2 responses (Graham et al, 1998).…”

Section: Discussionmentioning

confidence: 99%

Intranasal immunization with liposome-encapsulated plasmid DNA encoding influenza virus hemagglutinin elicits mucosal, cellular and humoral immune responses

Wang

Christopher

Nagata

et al. 2004

Journal of Clinical Virology

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In vivo protein expression and immune responses generated by DNA vaccines expressing mycobacterial antigens fused with a reporter protein

Cited by 7 publications

References 21 publications

Generation and characterization of a trackable plant-made influenza H5 virus-like particle (VLP) containing enhanced green fluorescent protein (eGFP)

Generation and characterization of a trackable plant-made influenza H5 virus-like particle (VLP) containing enhanced green fluorescent protein (eGFP)

Combined Triggering of Dendritic Cell Receptors Results in Synergistic Activation and Potent Cytotoxic Immunity

Intranasal immunization with liposome-encapsulated plasmid DNA encoding influenza virus hemagglutinin elicits mucosal, cellular and humoral immune responses

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