Department of Parasitology, College of Veterinary Medicine, Sichuan Agricultural University, Chengdu, China.
Department of Chemistry, College of Life and Basic Science, Sichuan Agricultural University, Chengdu, China.
Front Immunol. 2023 Jan 12;13:1037949. doi: 10.3389/fimmu.2022.1037949. eCollection 2022.
One of the most common rabbits coccidia species, is mainly parasitic in the ileal and jejunal epithelial cells. infection can affect the growth performance of rabbits or cause other secondary diseases. Traditional methods of anticoccidial treatment typically result in drug resistance and drug residue. Therefore, vaccination is a promising alternative. Gametocyte antigen 56 (GAM56) and rhoptry kinase family proteins (ROPs) are involved in oocyst wall formation and parasite invasion, respectively. A virulence factor, ROP17 contains a serine/threonine kinase catalytic domain. In this study, recombinant GAM56 (rGAM56) and ROP17 (rROP17) proteins were obtained from a prokaryotic expression system and their reactogenicity was investigated with immunoblotting. To assess the potential of rGAM56 and rROP17 as coccidiosis vaccines, New Zealand White rabbits were subcutaneously immunized with 100 μg rGAM56 (rGC group) or rROP17 (rRC group) twice at 2-week intervals followed by homologous oocyst challenge. The rabbit serum was collected weekly to detect the specific antibody levels. The cytokine levels of pre-challenge serum were measured by enzyme-linked immunosorbent assay and the rabbits were observed and recorded post-challenge for the onset of clinical symptoms. The weight gain, oocyst output, and feed conversion ratio were calculated at the end of the experiment. The results showed that both rGAM56 and rROP17 had good reactogenicity. The rGAM56- or rROP17-immunized rabbits had milder clinical symptoms and feed conversion ratios of 3.27:1 and 3.37:1, respectively. The rGAM56-immunized rabbits had 81.35% body weight gain and 63.85% oocyst output reduction; the rROP17-immunized rabbits had 79.03% body weight gain and 80.10% oocyst output reduction. The ACI of rGC and rRC groups were 162.35 and 171.03, respectively. The specific antibody levels increased rapidly after immunization. Significantly increased interleukin (IL)-2, interferon (IFN)-γ, and IL-17 levels were evident in the rGC and rRC groups (p < 0.05). The rGAM56 and rROP17 elicited humoral and cellular responses, which protected against infection in rabbits. Thus, rGAM56 and rROP17 are potential vaccine candidates against , and rROP17 performed better than rGAM56.
一种最常见的兔球虫种,主要寄生在回肠和空肠的上皮细胞中。感染会影响兔子的生长性能或导致其他继发疾病。传统的抗球虫药物治疗方法通常会导致药物耐药性和药物残留。因此,疫苗接种是一种很有前途的替代方法。配子细胞抗原 56(GAM56)和伸头激酶家族蛋白(ROPs)分别参与卵囊壁形成和寄生虫入侵。一种毒力因子,ROP17 含有丝氨酸/苏氨酸激酶催化结构域。在这项研究中,重组 GAM56(rGAM56)和 ROP17(rROP17)蛋白从原核表达系统中获得,并通过免疫印迹法研究其反应原性。为了评估 rGAM56 和 rROP17 作为球虫病疫苗的潜力,新西兰白兔用 100 μg rGAM56(rGC 组)或 rROP17(rRC 组)进行两次皮下免疫,间隔 2 周,然后进行同源卵囊攻击。每周采集兔血清检测特异性抗体水平。通过酶联免疫吸附试验测量预攻毒血清的细胞因子水平,并观察和记录攻毒后的临床症状出现情况。实验结束时计算体重增加、卵囊排出量和饲料转化率。结果表明,rGAM56 和 rROP17 均具有良好的反应原性。rGAM56 或 rROP17 免疫的兔子的临床症状较轻,饲料转化率分别为 3.27:1 和 3.37:1。rGAM56 免疫的兔子体重增加 81.35%,卵囊排出量减少 63.85%;rROP17 免疫的兔子体重增加 79.03%,卵囊排出量减少 80.10%。rGC 和 rRC 组的 ACI 分别为 162.35 和 171.03。免疫后抗体水平迅速升高。rGC 和 rRC 组的白细胞介素(IL)-2、干扰素(IFN)-γ和 IL-17 水平显著升高(p<0.05)。rGAM56 和 rROP17 引起了体液和细胞反应,对兔感染有保护作用。因此,rGAM56 和 rROP17 是兔球虫病的潜在疫苗候选物,rROP17 的效果优于 rGAM56。