Carrier protein influences immunodominance of a known epitope: implication in peptide vaccine design.
DIR@IMTECH: CSIR-Institute of Microbial Technology
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Title |
Carrier protein influences immunodominance of a known epitope: implication in peptide vaccine design.
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Creator |
Ghosh, Moumita
Solanki, Ashish K Roy, Koushik Dhoke, Reema R Ashish, . Roy, Syamal |
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Subject |
QR Microbiology
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Description |
We investigated how the processing of a given antigen by antigen presenting cells (APC) is dictated by the conformation of the antigen and how this governs the immunodominance hierarchy. To address the question, a known immunodominant sequence of bacteriophage lambda repressor N-terminal sequence 12-26 [λR(12-26)] was engineered at the N and C termini of a heterologous leishmanial protein, Kinetoplastid membrane protein-11 (KMP-11); the resulting proteins were defined as N-KMP-11 and C-KMP-11 respectively. The presence of λR(12-26) in N-KMP-11 and C-KMP-11 was established by western blot analysis with antibody to λR(12-26) peptide. N-KMP-11 but not C-KMP-11 could stimulate the anti λR(12-26) T-cell clonal population very efficiently in the presence of APCs. Priming of BALB/c mice with N-KMP-11 or C-KMP-11 generated similar levels of anti-KMP-11 IgG, but anti-λR(12-26) specific IgG was observed only upon priming with N-KMP-11. Interestingly, uptake of both N-KMP-11 and C-KMP-11 by APCs was similar but catabolism of N-KMP-11 but not C-KMP-11 was biphasic and fast at the initial time point. Kratky plots of small angle X-ray scattering showed that while N-KMP-11 adopts flexible Gaussian type of topology, C-KMP-11 prefers Globular nature. To show that KMP-11 is not unique as a carrier protein, an epitope (SPITBTNLBTMBK) of Plasmodium yoelii (PY) apical membrane protein 1[AMA-1 (136-148)], is placed at the C and N terminals of a dominant T-cell epitope of ovalbumin protein OVA(323-339) and the resulting peptides are defined as PY-OVA and OVA-PY respectively. Interestingly, only OVA-PY could stimulate anti-OVA T-cells and produce IgG response upon priming of BALB/c mice with it. Thus for rational design of peptide vaccine it is important to place the dominant epitope appropriately in the context of the carrier protein.
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Publisher |
Elsevier Science
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Date |
2013-09-23
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Type |
Article
PeerReviewed |
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Relation |
https://www.sciencedirect.com/science/article/pii/S0264410X13010104?via%3Dihub
http://crdd.osdd.net/open/2438/ |
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Identifier |
Ghosh, Moumita and Solanki, Ashish K and Roy, Koushik and Dhoke, Reema R and Ashish, . and Roy, Syamal (2013) Carrier protein influences immunodominance of a known epitope: implication in peptide vaccine design. Vaccine, 31 (41). pp. 4682-8. ISSN 1873-2518
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