Orientation of Antigen Display on Self-Assembling Protein Nanoparticles Influences Immunogenicity

Schneider, Cosette G.; Taylor, Justin A.; Sibilo, Michael Q.; Miura, Kazutoyo; Mallory, Katherine L.; Mann, Christopher; Karch, Christopher P.; Beck, Zoltán; Matyas, Gary R.; Long, Carole A.; Bergmann‐Leitner, Elke S.; Burkhard, Peter; Angov, Evelina

doi:10.3390/vaccines9020103

Cited by 11 publications

(12 citation statements)

References 92 publications

(117 reference statements)

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“…Combination of computational and structural biology allows the rational design of the well-oriented display of the most protective epitopes of a pathogen. Interestingly, orientation of antigen display on self-assembling protein nanoparticles has been shown to influence immunogenicity [ 43 ]. We cannot exclude that, based on the antigen investigated, retaining a native-like, unconstrained structure, allowing conformational epitopes to form, could have a positive impact on the immune response elicited and, actually, this could be critical for certain antigens.…”

Section: Discussionmentioning

confidence: 99%

Investigating the Role of Antigen Orientation on the Immune Response Elicited by Neisseria meningitidis Factor H Binding Protein on GMMA

Alfini

Brunelli²,

Bartolini³

et al. 2022

Vaccines

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GMMA are outer membrane vesicles (OMVs) released from Gram-negative bacteria genetically modified to enhance OMVs formation that have been shown to be optimal systems to enhance immunogenicity of protein antigens. Here, we selected Neisseria meningitidis factor H binding protein (fHbp) and used the conjugation chemistry as a tool to alter antigen orientation on GMMA. Indeed, fHbp was randomly linked to GMMA or selectively attached via the N-terminus to mimic native presentation of the protein on the bacterial surface. Interestingly, protein and peptide array analyses confirmed that antibodies induced by the selective and the random conjugates showed a pattern very similar to fHbp natively expressed on bacterial surfaces or to the recombinant protein mixed with GMMA, respectively. However, the two conjugates elicited antibodies with similar serum bactericidal activity against meningococcal strains, superior to the protein alone or physically mixed with GMMA. Presentation of fHbp on GMMA strongly enhances the functional immune response elicited by the protein but its orientation on the bacterial surface does not have an impact. This study demonstrates the flexibility of the GMMA platform as a display and delivery system for enhancing antigen immunogenicity and further supports the use of such promising technology for the development of effective vaccines.

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Section: Discussionmentioning

confidence: 99%

Investigating the Role of Antigen Orientation on the Immune Response Elicited by Neisseria meningitidis Factor H Binding Protein on GMMA

Alfini

Brunelli²,

Bartolini³

et al. 2022

Vaccines

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“…To determine antibodies specific to PfCelTOS, mice were bled via lateral tail veins and blood samples were collected prior to each immunization and the day of challenge. ELISA was performed as previously described ( 16 , 36 ). Briefly, 96-well 2HB Immunolon plates (Thermo Scientific, Waltham, MA) were coated with 100µL/well of 25ng/well of rPfCelTOS in 1X PBS, pH 7.4 (Quality Biological, Gaithersburg, MD) and incubated overnight at 4°C in a humidified chamber.…”

Section: Methodsmentioning

confidence: 99%

“…Densitometric detection of proteins in Western blots were quantified with ImageJ software (35). The 10ng recombinant PfCelTOS (rPfCelTOS) protein served as a relative control for quantification.…”

Section: Densitometry Analysis Of Western Blots Using Imagejmentioning

confidence: 99%

“…A functional signal sequence on secreted proteins has a role in protecting mRNA in the cytosol from degradation and the lack of such here, may account for the reduced protein translation levels for the pfceltos No-SS mRNA transcript. To estimate protein expression profiles of pfceltos mRNA's incorporating different signal sequences, relative density plots based on the 10ng rPfCelTOS reference standard, were analyzed using ImageJ, a Java-based image processing program, on Western blots (35). Differences in protein migration observed between the E. coli expressed reference protein and the in vitro translated protein is approximately 2KDa and can be accounted for by the inclusion of a 6-His+linker sequence on the rPfCelTOS.…”

Section: Different Signal Sequences Do Not Alter Translation and Cell...mentioning

confidence: 99%

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Exploring in vitro expression and immune potency in mice using mRNA encoding the Plasmodium falciparum malaria antigen, CelTOS

et al. 2022

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The secreted malarial protein, Cell-Traversal protein for Ookinetes and Sporozoites (CelTOS), is highly conserved among Plasmodium species, and plays a role in the invasion of mosquito midgut cells and hepatocytes in the vertebrate host. CelTOS was identified as a potential protective antigen based on a proteomic analysis, which showed that CelTOS stimulated significant effector T cells producing IFN-γ in peripheral blood mononuclear cells (PBMCs) from radiation attenuated sporozoite-immunized, malaria-naïve human subjects. In a rodent malaria model, recombinant full-length CelTOS protein/adjuvant combinations induced sterile protection, and in several studies, functional antibodies were produced that had hepatocyte invasion inhibition and transmission-blocking activities. Despite some encouraging results, vaccine approaches using CelTOS will require improvement before it can be considered as an effective vaccine candidate. Here, we report on the use of mRNA vaccine technology to induce humoral and cell-mediated immune responses using this antigen. Several pfceltos encoding mRNA transcripts were assessed for the impact on protein translation levels in vitro. Protein coding sequences included those to evaluate the effects of signal sequence, N-glycosylation on translation, and of nucleoside substitutions. Using in vitro transfection experiments as a pre-screen, we assessed the quality of the expressed CelTOS target relative to the homogeneity, cellular localization, and durability of expression levels. Optimized mRNA transcripts, which demonstrated highest protein expression levels in vitro were selected for encapsulation in lipid nanoparticles (LNP) and used to immunize mice to assess for both humoral and cellular cytokine responses. Our findings indicate that mRNA transcripts encoding pfceltos while potent for inducing antigen-specific cellular cytokine responses in mice, were less able to mount PfCelTOS-specific antibody responses using a two-dose regimen. An additional booster dose was needed to overcome low seroconversion rates in mice. With respect to antibody fine specificities, N-glycosylation site mutated immunogens yielded lower immune responses, particularly to the N-terminus of the molecule. While it remains unclear the impact on CelTOS antigen as immunogen, this study highlights the need to optimize antigen design for vaccine development.

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“…This malaria vaccine will be the first to be widely implemented, and thus, it will allow the research community to observe, for the first time, the possible benefits as well as effects on local parasite genetics and dynamics, disease prevalence, and transmission that can result from the introduction of a subunit malaria vaccine based on an immunodominant protein representing a single species and strain [ 287 , 288 ]. This achievement has and will continue to pave the way for testing of other currently available and future-generation vaccine candidates [ 280 , 281 , 289 – 293 ].…”

Section: Twenty-first Century—turning Point In Malaria Researchmentioning

confidence: 99%

Systems biology of malaria explored with nonhuman primates

Galinski

2022

Malar J

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Abstract“The Primate Malarias” book has been a uniquely important resource for multiple generations of scientists, since its debut in 1971, and remains pertinent to the present day. Indeed, nonhuman primates (NHPs) have been instrumental for major breakthroughs in basic and pre-clinical research on malaria for over 50 years. Research involving NHPs have provided critical insights and data that have been essential for malaria research on many parasite species, drugs, vaccines, pathogenesis, and transmission, leading to improved clinical care and advancing research goals for malaria control, elimination, and eradication. Whilst most malaria scientists over the decades have been studying Plasmodium falciparum, with NHP infections, in clinical studies with humans, or using in vitro culture or rodent model systems, others have been dedicated to advancing research on Plasmodium vivax, as well as on phylogenetically related simian species, including Plasmodium cynomolgi, Plasmodium coatneyi, and Plasmodium knowlesi. In-depth study of these four phylogenetically related species over the years has spawned the design of NHP longitudinal infection strategies for gathering information about ongoing infections, which can be related to human infections. These Plasmodium-NHP infection model systems are reviewed here, with emphasis on modern systems biological approaches to studying longitudinal infections, pathogenesis, immunity, and vaccines. Recent discoveries capitalizing on NHP longitudinal infections include an advanced understanding of chronic infections, relapses, anaemia, and immune memory. With quickly emerging new technological advances, more in-depth research and mechanistic discoveries can be anticipated on these and additional critical topics, including hypnozoite biology, antigenic variation, gametocyte transmission, bone marrow dysfunction, and loss of uninfected RBCs. New strategies and insights published by the Malaria Host–Pathogen Interaction Center (MaHPIC) are recapped here along with a vision that stresses the importance of educating future experts well trained in utilizing NHP infection model systems for the pursuit of innovative, effective interventions against malaria.

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Orientation of Antigen Display on Self-Assembling Protein Nanoparticles Influences Immunogenicity

Cited by 11 publications

References 92 publications

Investigating the Role of Antigen Orientation on the Immune Response Elicited by Neisseria meningitidis Factor H Binding Protein on GMMA

Investigating the Role of Antigen Orientation on the Immune Response Elicited by Neisseria meningitidis Factor H Binding Protein on GMMA

Exploring in vitro expression and immune potency in mice using mRNA encoding the Plasmodium falciparum malaria antigen, CelTOS

Systems biology of malaria explored with nonhuman primates

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