非洲猪瘟病毒P72蛋白合成肽疫苗的构建及其免疫效力评估

doi:10.16431/j.cnki.1671-7236.2022.08.024

Abstract

Abstract: 【Objective】 The aim of this study was to construct a synthetic peptide vaccine based on protein P72 in African swine fever virus (ASFV) strain CAS19-01/2019(GenBank accession No.:MIN172368.1),and evaluate its immune efficiency by immunizing mice.【Method】 The physicochemical properties and structural information of P72 protein were analyzed by programs of ProtParam and SOPMA.The significant epitopes in T and B lymphocytes were screened by ABCpred,SVMtrip and IEDCB.The synthesized peptides were injected intramuscularly into mice with Freund's adjuvant.Antibodies against the synthesized peptides,T lymphocyte subsets,lymphocyte proliferation,cytokines contents of interleukin 4(IL-4),IL-2,interferon-γ (IFN-γ) and immunoglobulin (IgG) were detected using serum and other tissue from the immunized mice,and the immune efficacy of the synthetic peptide was evaluated from the perspectives of humoral immunity and cellular immunity.【Result】 Comprehensive analysis showed that P72 protein was a stable hydrophilic protein.In secondary structure,alpha helix,beta turn,extended strand and random coil accounted for 19.35%,5.42%,25.08% and 50.15%,respectively.Eight dominant epitopes of protein P72 were selected by comprehensive software analysis,including T cell epitopes 626-634,520-528,298-306,203-211 amino acids,and B cell epitopes 587-606,232-251,110-129,39-58 amino acids. Two peptides named P72-1 and P72-2 were synthesized by integrating the dominant epitopes together.The specific antibodies against the synthesized peptides P72-1 and P72-2 were determined in serum of mice after 14 days of the first immunization,reached the highest value on the 28th day after the first immunization,and highest antibody titers were 1:25 600 and 1:12 800,respectively.The value of CD4⁺/CD8⁺ of T lymphocyte subsets in immunized mice were significantly increased (P<0.05).The spleen lymphocyte proliferation test showed that the number of lymphocytes was enhanced in both groups (P<0.01).Cytokines contents of IL-4,IL-2 and IFN-γ were extremely significantly improved (P<0.01).【Conclusion】 In this study,two synthetic peptide vaccines were successfully developed.P72-2 was higher than P72-1 in immune efficacy,and both could produce high level of specific antibodies,stimulate the proliferation of lymphocytes,induce the production of cytokines IL-4,IL-2 and IFN-γ.This study laid a technical foundation for the development of African swine fever vaccine.

Key words: African swine fever virus (ASFV); P72 protein; antigen epitopes; synthetic peptide vaccine

CLC Number:

S852.65⁺1

XU Yaling, ZHANG Jihui, NIU Xi, LI Sheng, HUANG Shihui, RAN Xueqin, WANG Jiafu. Construction of Synthetic Peptide Vaccine of African Classical Swine Fever Virus P72 Protein and Evaluation of Its Immune Efficacy[J]. China Animal Husbandry and Veterinary Medicine, 2022, 49(8): 3083-3090.

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Construction of Synthetic Peptide Vaccine of African Classical Swine Fever Virus P72 Protein and Evaluation of Its Immune Efficacy

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