Fusion of the Mycobacterium tuberculosis Antigen 85A to an Oligomerization Domain Enhances Its Immunogenicity in Both Mice and Non-Human Primates

Spencer, Alexandra J.; Hill, Fergal; Honeycutt, Jared; Cottingham, Matthew G.; Bregu, Migena; Rollier, Christine S.; Furze, Julie; Draper, Simon J.; Søgaard, Karen C.; Gilbert, Sarah C.; Wyllie, David; Hill, Adrian V. S.

doi:10.1371/journal.pone.0033555

Cited by 43 publications

(52 citation statements)

References 41 publications

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“…There is also potential for free-peptide to be more readily degraded by proteases, lowering peptide activity, however activity was comparable to another study9. In addition to Cpn10, the benefits of peptide display on heptamers has been demonstrated by fusing an oligomerisation domain to the Mycobacterium tuberculosis antigen 85A14. In this example the presentation of the 85A antigen in heptamer form improved molecule activity by enhancing immunogenicity, specifically the induction of T cell responses in both mice and non-human primate studies.…”

Section: Discussionmentioning

confidence: 81%

Beyond Antibodies: Development of a Novel Protein Scaffold Based on Human Chaperonin 10

Alsultan

Chin

Howard

et al. 2016

Sci Rep

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Human Chaperonin 10 (hCpn10) was utilised as a novel scaffold for presenting peptides of therapeutic and diagnostic significance. Molecular dynamic simulations and protein sizing analyses identified a peptide linker (P1) optimal for the formation of the quarternary hCpn10 heptamer structure. hCpn10 scaffold displaying peptides targeting Factor VIIa (CE76-P1) and CD44 (CP7) were expressed in E. coli. Functional studies of CE76-P1 indicated nanomolar affinity for Factor VIIa (3 nM) similar to the E-76 peptide (6 nM), with undetectable binding to Factor X. CE76-P1 was a potent inhibitor of FX activity (via inhibition of Factor VIIa) and prolonged clot formation 4 times longer than achieved by E-76 peptide as determined by prothrombin time (PT) assays. This improvement in clotting function by CE76-P1, highlights the advantages of a heptamer-based scaffold for improving avidity by multiple peptide presentation. In another example of hCPn10 utility as a scaffold, CP7 bound to native CD44 overexpressed on cancer cells and bound rCD44 with high affinity (KD 9.6 nM). The ability to present various peptides through substitution of the hCpn10 mobile loop demonstrates its utility as a novel protein scaffold.

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Section: Discussionmentioning

confidence: 81%

Beyond Antibodies: Development of a Novel Protein Scaffold Based on Human Chaperonin 10

Alsultan

Chin

Howard

et al. 2016

Sci Rep

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“…Detection of anti-Ag85A IgG levels in DDA-MPL+Ag85A was compared to those of other groups (a), the ratio of IgG2b/IgG1 was calculated by the titre of IgG2b and IgG1 (b). vaccine against TB for use in humans is BCG, which has been in use since 1921 (Spencer et al, 2012); however, its efficacy is highly variable, and particularly so for protection against adult pulmonary TB. Therefore, there is an urgent need for a vaccine with better protection against adult pulmonary disease than that afforded by BCG.…”

Section: Discussionmentioning

confidence: 99%

Enhanced immune responses in mice to combined immunization with Mycobacterium tuberculosis Ag85A and DDA/MPL

Zhao¹,

Tan²,

Xu³

et al. 2014

Afr. J. Microbiol. Res.

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The tuberculosis (TB) Bacillus Calmette-Guerin (BCG) vaccine, which is based on Mycobacterium bovis, has variable protective efficacy in humans; therefore, safe and effective vaccines are urgently required to prevent this disease. The antigen 85A from Mycobacterium tuberculosis (M. tuberculosis) is a prime target for immunity in the early phase of tuberculosis in various animal models. The purpose of this study was to confirm whether Ag85A could elicit protective immune responses in mice. Dimethyldioctadecylammonium (DDA), an adjuvant with Th1-promoting activities and monophosphoryl lipid A (MPL), an immunostimulatory component that has strong adjuvant activity for both cellular and humoral immune responses, has been used as the adjuvant to study the biological and immunological characteristics of tuberculosis proteins in the researches previously. The gene coding Ag85A (fbpA), was cloned into a pET-30a(+) prokaryotic expression vector, and the induced amino acid sequence corresponds to a 33 kDa protein, which was confirmed by mass spectrometry and western blots. Mice were subcutaneously immunized with Ag85A protein emulsified with DDA and MPL and the results showed that the 85A antigen, when combined with these adjuvants, elicited strong Ag85A-specific T-cell responses and humoral responses as compared with vaccination with BCG. Based on the fact that this vaccine combination induced strong antigen-specific immune responses, it is a prime candidate as a component of a future TB vaccine.

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“…In mice, this construct provided significant protection against challenge although not to the same extent as the control BCG vaccine. Similarly, antigen multimerization has been used for enhancing the immunogenicity of proteins and a fusion of Ag85A to IMX313, a hybrid avian oligomerization domain, showed enhanced CD4 and CD8 T cell responses when used in DNA and MVA vaccine constructs [97]. In order to induce combined CD4 and CD8 T cell responses, a formulation of IL-12 delivered by the heamagglutinin virus of Japan-envelope and –liposomes has also been used.…”

Section: Preclinical Adjuvant Candidatesmentioning

confidence: 99%

Novel adjuvant formulations for delivery of anti-tuberculosis vaccine candidates

Agger

2016

Advanced Drug Delivery Reviews

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There is an urgent need for a new and improved vaccine against tuberculosis for controlling this disease that continues to pose a global health threat. The current research strategy is to replace the present BCG vaccine or boost BCG-immunity with subunit vaccines such as viral vectored-or protein-based vaccines. The use of recombinant proteins holds a number of production advantages including ease of scalability, but requires an adjuvant inducing cell-mediated immune responses. A number of promising novel adjuvant formulations have recently been designed and show evidence of induction of cellular immune responses in humans. A common trait of effective TB adjuvants including those already in current clinical testing is a two-component approach combining a delivery system with an appropriate immunomodulator. This review summarises the status of current TB adjuvant research with a focus on the division of labor between delivery systems and immunomodulators. Graphical Abstract

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Fusion of the Mycobacterium tuberculosis Antigen 85A to an Oligomerization Domain Enhances Its Immunogenicity in Both Mice and Non-Human Primates

Cited by 43 publications

References 41 publications

Beyond Antibodies: Development of a Novel Protein Scaffold Based on Human Chaperonin 10

Beyond Antibodies: Development of a Novel Protein Scaffold Based on Human Chaperonin 10

Enhanced immune responses in mice to combined immunization with Mycobacterium tuberculosis Ag85A and DDA/MPL

Novel adjuvant formulations for delivery of anti-tuberculosis vaccine candidates

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