Institute of Health Sciences (ICS), Federal University of Bahia (UFBA), Salvador, Brazil; College of Animal Sciences, Faculty of Agricultural Sciences and Natural Resources, University of the Llanos, Villavicencio, Colombia.
Institute of Health Sciences (ICS), Federal University of Bahia (UFBA), Salvador, Brazil.
Vaccine. 2022 Feb 7;40(6):912-923. doi: 10.1016/j.vaccine.2021.12.052. Epub 2022 Jan 7.
Toxocariasis is a neglected parasitic zoonosis of global importance. The development of a formulation that can be used as a vaccine would help the definitive control of the infection. Preclinical studies selected two recombinant T. canis proteins (rTcVcan and rTcCad) which significantly protected mice against larval migration. In the present work, these proteins plus three adjuvants (Alhydrogel®, PAM3CSK4®, and Quil-A®) were used to immunize mice against toxocariasis; blood samples were collected three times to measure IgG (total, IgG1, IgG2a), IgA, and IgE via indirect ELISA. Cytokines (IL-5, TNF-α, and IL-10) were measured in splenocytes supernatant, and T. canis larvae were quantified in tissues. The best protein + adjuvant pair found (rTVcan + QuialA®) was then used to immunize T. canis-free puppies (n = 18) that were experimentally infected with T. canis and T. canis naturally-infected puppies (n = 6). Immunoglobulin (IgA, IgE, IgG, IgG1, and IgG2a), parasite load (eggs in feces), number of expelled adults and eggs extracted from the female uterus, and their fertility percentages were analyzed. In mice, it was observed a highly significant reduction (73%) of tissue larvae, a mixed cytokine profile (Th/Th), and anti-T. canis antibody titers (IgG, IgG1, IgG2a) using rTVcan + QuialA® mix. In canines, rTVcan + QuialA® promoted reduction in the parasite eggs in feces (95%) and eggs reduction obtained from the uteri of pharmacologically expelled adult females (58.38%). In our knowledge this is the first canine clinical trial of a vaccine with T. canis recombinant proteins. The formulation used has been shown to efficiently stimulate the production of antibodies against infection by T. canis. In the canine, a significant reduction in the number of eggs expelled by the experimental animals that received the formulation prophylactically was evidenced. Future tests should be developed to evaluate the duration of the protective effect and analyze other immune pathways that could be stimulated by the formulation used.
旋毛虫病是一种被忽视的具有全球重要性的寄生虫性动物传染病。开发一种可用于疫苗的制剂将有助于对感染的最终控制。临床前研究选择了两种重组犬旋毛虫蛋白(rTcVcan 和 rTcCad),它们能显著保护小鼠免受幼虫迁移的影响。在本工作中,这些蛋白与三种佐剂(Alhydrogel®、PAM3CSK4®和 Quil-A®)一起用于免疫小鼠以抵抗旋毛虫病;采集三次血样,通过间接 ELISA 测量 IgG(总、IgG1、IgG2a)、IgA 和 IgE。测量脾细胞上清液中的细胞因子(IL-5、TNF-α 和 IL-10),并在组织中定量检测犬旋毛虫幼虫。发现的最佳蛋白+佐剂组合(rTVcan+QuialA®)随后用于免疫无犬旋毛虫的幼犬(n=18),这些幼犬经实验感染犬旋毛虫和自然感染犬旋毛虫的幼犬(n=6)。分析免疫球蛋白(IgA、IgE、IgG、IgG1 和 IgG2a)、寄生虫负荷(粪便中的虫卵)、从雌性子宫中排出的成虫数量和从雌性子宫中提取的虫卵以及它们的生育率百分比。在小鼠中,使用 rTVcan+QuialA®混合物观察到组织幼虫数量显著减少(73%)、混合细胞因子谱(Th/Th)和抗犬旋毛虫抗体滴度(IgG、IgG1、IgG2a)。在犬中,rTVcan+QuialA®促进了粪便中寄生虫卵的减少(95%)和从药理学上排出的成年雌性子宫中获得的卵的减少(58.38%)。据我们所知,这是首例使用犬旋毛虫重组蛋白的犬临床试验。所使用的制剂已被证明能有效地刺激针对犬旋毛虫感染的抗体产生。在犬中,预防性给予该制剂的实验动物排出的卵数量显著减少。未来的测试应该开发出来,以评估保护作用的持续时间,并分析该制剂可能刺激的其他免疫途径。