creators_name: Ghosh, Moumita
creators_name: Solanki, Ashish K
creators_name: Roy, Koushik
creators_name: Dhoke, Reema R
creators_name: Ashish, .
creators_name: Roy, Syamal
type: article
datestamp: 2019-09-05 13:47:56
lastmod: 2019-09-05 13:47:56
metadata_visibility: show
title: Carrier protein influences immunodominance of a known epitope: implication in peptide vaccine design.
ispublished: pub
subjects: QR
keywords: AMA-1; APC; Antigen processing; Antigen structure; C-KMP-11; Endoproteolysis; Immunodominance; KMP-11; N-KMP-11; OVA((323–339)); PY; an epitope of AMA-1((136–148)); antigen presenting cell; dominant T-cell epitope of ovalbumin protein; immunodominant sequence of bacteriophage lambda repressor N-terminal sequence; kinetoplastid membrane protein-11; plasmodium yoelii apical membrane protein 1; λR((12–26)); λR((12–26)) engineered at the C termini of KMP-11; λR((12–26)) engineered at the N termini of KMP-11
note: Copyright of this article belongs to Elsevier Science.
abstract: 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.
date: 2013-09-23
date_type: published
publication: Vaccine
volume: 31
number: 41
publisher: Elsevier Science
pagerange: 4682-8
refereed: TRUE
issn: 1873-2518
official_url: https://www.sciencedirect.com/science/article/pii/S0264410X13010104?via%3Dihub
citation:   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