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免疫功能正常的仔猪对猪等孢球虫感染的抗体和细胞因子反应。

Antibody and cytokine response to Cystoisospora suis infections in immune-competent young pigs.

机构信息

Institute of Parasitology, Department of Pathobiology, University of Veterinary Medicine Vienna, Veterinaerplatz 1, 1210, Vienna, Austria.

HIPRA, Amer, Spain.

出版信息

Parasit Vectors. 2018 Jul 4;11(1):390. doi: 10.1186/s13071-018-2974-6.

Abstract

BACKGROUND

To date, investigations on the immune response to Cystoisospora suis infections focused on suckling piglets, the age group clinically most affected. Actively immunizing piglets is unfeasible due to their immature immune system and the typically early infection in the first days after birth. Therefore, understanding and possibly enhancing the immune response of immune-competent animals is the prerequisite to develop a passive immunization strategy for piglets which currently rely on very limited treatment options.

METHODS

To investigate antibody and cytokine responses of immune-competent animals and the impact of the oral immunization protocol on their immune response, growers with unknown previous exposure to C. suis (10-11 weeks-old) were infected one or three times with different doses (600 and 6000 or 200 and 2000, respectively) of C. suis oocysts, and compared to uninfected controls. Oocyst excretion was evaluated, and blood and intestinal mucus antibody titers were determined by IFAT. Systemic production of Th1, Th2, inflammatory and regulatory cytokines was determined in different immune compartments at mRNA and (after stimulation with a recombinant merozoite-protein) at protein level by PCR and multiplex fluorescent immunoassay, respectively.

RESULTS

Infection generated significantly increased serum IgA and IgG levels against C. suis sporozoites and merozoites, irrespective of infection mode, with IgG against merozoites showing the strongest increase. No clinical signs and only occasional excretion were observed. The systemic cytokine response to C. suis was only weak. Nonetheless, in white blood cells, IL-4, IL-6 and IL-10 mRNA-levels significantly increased after infection, whereas IFN-ɣ, IL-2 and TGF-β expression tended to decrease. In mesenteric lymph nodes (MLN), IL-10 and TNF-α levels were elevated while splenic cytokine expression was unaltered upon infection. Stimulated MLN-derived lymphocytes from infected pigs produced slightly more IL-12 and less IFN-α than controls.

CONCLUSIONS

An infection and a subsequent systemic immune response can be induced in immune-competent animals by all evaluated infection models and growers can be used as models to mimic sow immunizations. The immune response to C. suis, although mild and with considerable variation in cytokine expression, was characterized by a Th2-associated and regulatory cytokine profile and antibody production. However, none of the parameters clearly stood out as a potential marker associated with protection. Antibody titers were significantly positively related with oocyst excretion and might thus serve as correlates for parasite replication or severity of infection.

摘要

背景

迄今为止,针对猪等孢球虫感染的免疫反应的研究主要集中在哺乳期仔猪上,这是临床上受感染最严重的年龄组。由于仔猪的免疫系统不成熟,并且通常在出生后几天内就会早期感染,因此,对免疫功能正常的动物的免疫反应进行理解和可能增强是为仔猪开发被动免疫策略的前提,而目前仔猪的治疗选择非常有限。

方法

为了研究免疫功能正常的动物的抗体和细胞因子反应以及口服免疫方案对其免疫反应的影响,将未接触过猪等孢球虫(10-11 周龄)的育肥猪分别用不同剂量(分别为 600 和 6000 或 200 和 2000 个卵囊)进行一次或三次感染,并与未感染的对照组进行比较。通过间接免疫荧光试验(IFAT)测定粪便和肠粘液中的抗体滴度,评估卵囊的排出情况。通过聚合酶链反应(PCR)和多色荧光免疫测定法(分别为在重组裂殖子蛋白刺激后),在不同的免疫部位测定 Th1、Th2、炎症和调节性细胞因子的系统产生。

结果

感染导致针对猪等孢球虫的裂殖子和孢子的血清 IgA 和 IgG 水平显著升高,无论感染方式如何,针对裂殖子的 IgG 升高最为显著。仅观察到偶尔的临床症状和偶发性的卵囊排出。针对猪等孢球虫的系统细胞因子反应较弱。尽管如此,在白细胞中,感染后 IL-4、IL-6 和 IL-10 的 mRNA 水平显著增加,而 IFN-ɣ、IL-2 和 TGF-β的表达则趋于降低。在肠系膜淋巴结(MLN)中,IL-10 和 TNF-α 水平升高,而脾脏的细胞因子表达在感染后没有变化。与对照组相比,感染猪的 MLN 来源的淋巴细胞产生的 IL-12 略多,IFN-α 略少。

结论

所有评估的感染模型都可以在免疫功能正常的动物中诱导感染和随后的全身免疫反应,并且育肥猪可以作为模拟母猪免疫接种的模型。尽管仔猪的免疫反应温和,细胞因子表达差异较大,但表现出 Th2 相关和调节性细胞因子的特征,以及抗体的产生。然而,没有任何参数明显成为与保护相关的潜在标志物。抗体滴度与卵囊排出量呈显著正相关,因此可能作为寄生虫复制或感染严重程度的相关指标。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/6900/6031197/9efc4d9e6c3e/13071_2018_2974_Fig1_HTML.jpg

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