Production, purification, and in vivo evaluation of a novel multiepitope peptide vaccine consisted of immunodominant epitopes of SYCP1 and ACRBP antigens as a prophylactic melanoma vaccine.

Production, purification, and in vivo evaluation of a novel multiepitope peptide vaccine consisted of immunodominant epitopes of SYCP1 and ACRBP antigens as a prophylactic melanoma vaccine.

Publication date: Sep 06, 2019

Melanoma cells are significantly resistance to the current treatments. Therefore, the best option for high-risk populations is prevention. Recently, many preventive cancer vaccines have been developed. In our previous study, several bioinformatic tools were employed for selection of the most immunodominant epitopes of acrosin binding protein (ACRBP) and synaptonemal complex protein 1 (SYCP1) antigens to design multiepitope DNA and peptide cancer vaccines. In the current study, the final construct of the multiepitope DNA vaccine was placed into a pcDNA3.1 vector and then, subcloned into a pET-28a (+) expression vector for transfecting BL21 E. coli strain. The recombinant multiepitope peptide vaccine, weighing 6.35 kDa, was purified by Fast protein liquid chromatography technique (FPLC) and detected by western blotting. Subsequently, C57BL/6 mice were immunized by a mixture of the peptide vaccine and incomplete Freund’s adjuvant (IFA) (four vaccinations with one-week intervals). Two weeks after the last vaccination, the serum levels of the peptide-specific IgG total, IgG2a, and IgG1 were measured by enzyme-linked immunosorbent assays (ELISA). Also, the immunized mice splenocytes efficacy for producing interleukin-4 (IL-4) and interferon-γ (IFN-γ) after stimulation with the peptide vaccine was evaluated. At last, the prophylactic effect of the peptide vaccine immunization was evaluated in B16-F10 murine melanoma model. The peptide vaccine immunization caused a significant increase in the serum levels of IgG1, IgG2a, and IgG2a. Also, the immunized mice splenocytes exhibited significantly higher ability to produce IL-4 (10-fold) and IFN-γ (16-fold) after stimulation with the peptide vaccine, in comparison with the PBS and IFA groups. The peptide immunized mice exhibited 50.2% and 43% decrease in the mean tumors’ volume in comparison with PBS and IFA groups. Also, the mean survival time for the peptide immunized mice was 33 +/- 1.3 days which was 5 and 6 days more than the PBS and IFA groups, respectively. The obtained results exhibit high efficacy of the designed multiepitope peptide vaccine for the immune system activation and anti-tumor prophylactic effects in the murine melanoma model.

Safavi, A., Kefayat, A., Sotoodehnejadnematalahi, F., Salehi, M., and Modarressi, M.H. Production, purification, and in vivo evaluation of a novel multiepitope peptide vaccine consisted of immunodominant epitopes of SYCP1 and ACRBP antigens as a prophylactic melanoma vaccine. 24055. 2019 Int Immunopharmacol (76):

Concepts Keywords
Adjuvant Murine melanoma
Antigens Decrease tumors
Bioinformatic Immunization
ELISA Immunotherapy
Enzyme Vaccinations
Epitopes Vaccination
IFA Medical specialties
IgG Medicine
IgG1 Health
IgG2a Peptide vaccine
Immune System Adjuvant
Immunization Immunization
Interferon Biotechnology
Interleukin 4
Liquid Chromatography
Melanoma
Murine
PBS
Peptide
Prophylactic
Recombinant
Serum
Tumor
Vaccination
Vaccine
Vector
Vivo
Western Blotting

Semantics

Type Source Name
pathway BSID Immune System
gene UNIPROT F10
gene UNIPROT FAM110A
gene UNIPROT IL4
drug DRUGBANK Binetrakin
disease DOID cancer
disease MESH cancer
pathway BSID Melanoma
disease DOID melanoma
disease MESH melanoma
gene UNIPROT ACRBP
gene UNIPROT SYCP1

Original Article

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