副猪革拉瑟菌影递送猪圆环病毒2型DNA二联疫苗的制备及小鼠免疫效果评价

doi:10.11843/j.issn.0366-6964.2024.03.030

Abstract

Abstract: The aim of this study was to develop a combined vaccine against porcine circovirus type 2 (PCV2) and Glaesserella parasuis (GPS), and to evaluate its immune protective effect in mice. We selected SH0165, a type 5 strain of GPS, as the vaccine strain. The G. parasuis ghosts were generated by chemically-induced lysis and the method is based on minimum inhibitory concentration (MIC) of sodium hydroxide (NaOH). Then, the ORF2 gene of PCV2 was cloned into the pEGFP-N1 vector. We developed a PCV2-GPS combined vaccine by using G. parasuis ghosts as vehicle for pEGFP-ORF2. Furthermore, four-week-old BALB/c mice were divided into commercial vaccine group, PCV2-GPS combined vaccine group, G. parasuis vaccine group, pEGFP-ORF2 group, PBS group, and blank group. The IgG antibody level against PCV2 and GPS, neutralization antibody against PCV2, PCV2 viral load of organs in mice, serum bactericidal activities, challenge protection efficacy, and level of cytokines (IFN-γ, IL-12) in serum were measured after immunization. The results showed that PCV2-GPS combined vaccine elicited significantly higher levels of IgG antibody response of PCV2 and GPS, neutralization antibody titer of PCV2, bactericidal activities and higher levels of IFN-γ than those of the control group. After PCV2 challenge, PCV2-GPS combined vaccine-immunized mice showed lower viral load in lymph nodes and lungs than the control group, and after challenge infection with GPS, immune protection efficacy of mice immunized was 70% (7/10). In conclusion, the PCV2-GPS combined vaccine significantly activated both humoral and cell-mediated immune responses, and inhibited the proliferation of PCV2 in mice, achieving a better protective effect in mice infected with GPS. The results provide biological materials and experimental data for the development of new, safe and efficient combined vaccines against porcine circovirus type 2 and Glaesserella parasuis.

Key words: porcine circovirus type 2, ORF2 gene, Glaesserella parasuis, bacterial ghost, mice, immunoprotection

CLC Number:

S855

WANG Kang, LIU Geyan, WANG Yu, YANG Zhen, TANG Xinwei, CAO Sanjie, HUANG Xiaobo, YAN Qigui, WU Rui, ZHAO Qin, DU Senyan, WEN Xintian, WEN Yiping. Preparation of Glaesserella parasuis Ghosts Vaccine Delivering Porcine Circovirus Type 2 DNA Vaccine and Evaluation of Its Immunoprotective Effect in Mice[J]. Acta Veterinaria et Zootechnica Sinica, 2024, 55(3): 1179-1191.

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Preparation of Glaesserella parasuis Ghosts Vaccine Delivering Porcine Circovirus Type 2 DNA Vaccine and Evaluation of Its Immunoprotective Effect in Mice

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