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首页> 外文期刊>Journal of Parasitology >REDUCED INFECTION AND PROTECTION FROM CLINICAL SIGNS OF CEREBRAL NEOSPOROSIS IN C57BL/6 MICE VACCINATED WITH RECOMBINANT MICRONEME ANTIGEN NCMIC1
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REDUCED INFECTION AND PROTECTION FROM CLINICAL SIGNS OF CEREBRAL NEOSPOROSIS IN C57BL/6 MICE VACCINATED WITH RECOMBINANT MICRONEME ANTIGEN NCMIC1

机译：接种重组细微克隆抗原NCMIC1的C57BL / 6小鼠的脑部新疾病的临床体征减少了感染和预防

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摘要

NcMIC1 is a 460 amino acid Neospora caninum microneme protein implicated in host cell adhesion and invasion processes. In this study, we assessed the potential protectivity of NcMIC1-based vaccination against experimental N. caninum infection in mice, employing both recombinant antigen vaccines and DNA vaccines. Recombinant NcMIC1 (recNcMIC1) was expressed in Escherichia coli as gluthatione-S-transferase-fusion protein. The corresponding NcMIC1 cDNA was cloned into the pcDNA3.1 expression plasmid (pcDNA-MIC1), and expression was checked in transfected Vero cells. Mice (10 animals/group) were vaccinated either with recNcMIC1 antigen suspended in Ribi-adjuvant (3 intraperitoneal injections), pcDNA-NcMIC1 (3 intramuscular injections), or pcDNA-NcMIC1 (twice intramuscularly), followed by 1 intraperitoneal recNcMIC1 antigen boost. Control groups included corresponding treatments with adjuvant, pcDNA3.1 without insert, and PBS (= infection control). All vaccinated and control groups were then challenged intraperitoneally with 2 × 106 N. caninum tachyzoites. Animals were inspected daily for a period of 3 wk postinfection (PI). At day 21, all animals were killed and assessed for infection. Before day 21 PI, clinical signs such as walking disorders, rounded back, apathy, and paralysis occurred in infection controls (50% of the mice), pcDNA and adjuvant controls (20% each), and the combined pcDNA-NcMIC1/recNcMIC1-treated group (30%). No clinical symptoms were observed in the recNcMIC1 and pcDNA-NcMIC1 vaccinated groups. All mice were positive for cerebral N. caninum infection as assessed by PCR of brain tissue. However, quantitative real-time PCR revealed that the infection intensity was significantly reduced in the group vaccinated with recNcMIC1 antigen. Immunohistochemistry confirmed these findings. In contrast, the infection intensity was highest in the group vaccinated with the pcDNA-NcMIC1/recNcMIC1 combination, indicating that the sequential application of the DNA vaccine and recombinant antigen had a deleterious effect. Serological analysis showed that only recNcMIC1-immunized animals generated detectable antibody levels recognizing native NcMIC1. Thus, of all protocols applied here, only recNcMIC1 vaccination appears to be suited to reduce cerebral infection in mice challenged with N. caninum tachyzoites.
机译：NcMIC1是一种460个氨基酸的犬新孢子虫微neme蛋白，与宿主细胞的粘附和侵袭过程有关。在这项研究中，我们使用重组抗原疫苗和DNA疫苗评估了基于NcMIC1的疫苗对小鼠实验性猪链球菌感染的潜在保护性。重组NcMIC1（recNcMIC1）在大肠杆菌中以谷胱甘肽S-转移酶融合蛋白的形式表达。将相应的NcMIC1 cDNA克隆到pcDNA3.1表达质粒（pcDNA-MIC1）中，并在转染的Vero细胞中检查表达。给小鼠（10只动物/组）接种悬浮在Ribi-佐剂中的recNcMIC1抗原（3次腹膜内注射），pcDNA-NcMIC1（3次肌肉内注射）或pcDNA-NcMIC1（2次肌肉内）接种，然后进行1次腹膜内recNcMIC1抗原加强免疫。对照组包括相应的佐剂，pcDNA3.1（无插入片段）和PBS（=感染对照）治疗。然后，所有疫苗接种组和对照组均用2×106 N. caninum tachyzoite腹膜内攻击。每天检查动物感染后3周（PI）的时间。在第21天，杀死所有动物并评估感染。在注射后第21天之前，感染控制（50％的小鼠），pcDNA和佐剂控制（每组20％）以及组合的pcDNA-NcMIC1 / recNcMIC1-发生了行走障碍，后退，冷漠和麻痹等临床症状。治疗组（30％）。在recNcMIC1和pcDNA-NcMIC1接种组中未观察到临床症状。通过脑组织的PCR评估，所有小鼠的脑犬新孢子虫感染均为阳性。然而，定量实时PCR显示，在接种recNcMIC1抗原的组中，感染强度显着降低。免疫组织化学证实了这些发现。相反，在接种了pcDNA-NcMIC1 / recNcMIC1组合的组中，感染强度最高，表明DNA疫苗和重组抗原的顺序施用具有有害作用。血清学分析表明，只有经recNcMIC1免疫的动物才能产生可​​识别天然NcMIC1的抗体水平。因此，在此处应用的所有方案中，只有recNcMIC1疫苗似乎适合减少犬新孢子虫速殖子攻击的小鼠的脑部感染。

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