Construction of HCV-polytope vaccine candidates harbouring immune-enhancer sequences and primary evaluation of their immunogenicity in BALB/c mice

Autor: Arash Memarnejadian, Farzin Roohvand, Mohammad Reza Aghasadeghi, Arash Arashkia, Sima Rafati
Přispěvatelé: Hepatitis & AIDS Dept., Réseau International des Instituts Pasteur (RIIP)-Institut Pasteur d'Iran, Réseau International des Instituts Pasteur (RIIP), Institut Pasteur d'Iran, Financially supported by Pasteur Institute of Iran., Réseau International des Instituts Pasteur (RIIP)-Réseau International des Instituts Pasteur (RIIP)
Rok vydání: 2009
Předmět:
HBsAg
T-Lymphocytes
Epitopes
T-Lymphocyte
Hepacivirus
MESH: Amino Acid Sequence
Epitope
Mice
0302 clinical medicine
Chlorocebus aethiops
MESH: Animals
MESH: Hepacivirus
Antigens
Viral
Mice
Inbred BALB C
0303 health sciences
biology
Viral Vaccine
ELISPOT
Immunogenicity
General Medicine
3. Good health
MESH: COS Cells
COS Cells
[SDV.IMM]Life Sciences [q-bio]/Immunology
Female
030211 gastroenterology & hepatology
MESH: Antigens
Viral
Molecular Sequence Data
MESH: Mice
Inbred BALB C
BALB/c
MESH: Epitopes
T-Lymphocyte
03 medical and health sciences
Antigen
MESH: Viral Vaccines
Virology
Genetics
Animals
Humans
Amino Acid Sequence
MESH: Mice
Molecular Biology
MESH: HLA-A Antigens
030304 developmental biology
MESH: Molecular Sequence Data
MESH: Humans
HLA-A Antigens
Viral Vaccines
biology.organism_classification
MESH: Cercopithecus aethiops
Molecular biology
MESH: T-Lymphocytes
MESH: Female
Minigene
Zdroj: Virus Genes
Virus Genes, Springer Verlag, 2010, 40 (1), pp.44-52. ⟨10.1007/s11262-009-0417-3⟩
ISSN: 1572-994X
0920-8569
DOI: 10.1007/s11262-009-0417-3
Popis: International audience; An efficient vaccine against hepatitis-C virus (HCV) infection requires vigorous and focused CD8(+) T-cell responses against viral antigens. Due to immunosuppressive effect of HCV antigens, polytope vaccines comprising the minimal CD8(+)CTL epitopes are of peculiar concern. Herein, to provide information for construction of efficient HCV polytope vaccine candidates, one H-2D(d) (E2(405-414):E(2)) and two HLA-A*0201 (E1(363-372):E(1) and Core(35-44):C)-restricted CD8(+) T-cell epitopes of HCV were selected. By employing number of in silico analyses, the E(2)E(1)C linear format was predicted as optimum epitope consecution and after amplification by SOEing-PCR, the corresponding DNA sequence was cloned in pcDNA3.1+ vector. To further evaluate the role of immune-enhancer elements, a universal T-helper epitope (PADRE), endoplasmic reticulum signal sequence (ERss) and hepatitis-B surface-antigen (HBsAg) gene were fused separately or in combination to the E(2)E(1)C minigene. In vitro analyses of polytopes by different DNA/protein-based assays demonstrated proper transcription/expression of constructs in transfected cells. Measurement of the HBsAg-mediated particle secretion by ELISA indicated lack of secretion in the related polytopes. Results of delayed-type hypersensitivity (DTH) as a preliminary in vivo analysis, and confirmatory ELISPOT assays showed the proper processing and presentation of H-2D(d)-restricted-E(2) epitope and approved the enhancing effect of PADRE and ERss sequences but not HBsAg for the immune responses against E(2) in immunized BALB/c mice. Our results pointed to the value of in silico predictions and application of immune-enhancer elements as well as DTH analysis for design and primary in vivo evaluation of HCV polytopes, prior to costly transgenic studies on immunogenicity of HLA-A*0201 epitopes.
Databáze: OpenAIRE
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