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评价粗提猪蛔虫成虫肠道匀浆诱导 BALB/c 小鼠抗猪蛔虫幼虫保护性 IgG 产生的作用。

Evaluation of crude adult Ascaris suum intestinal tract homogenate in inducing protective IgG production against A. suum larvae in BALB/c mice.

机构信息

College of Medicine, University of the Philippines Manila, Pedro Gil Street, Malate, Manila, 1000, Philippines.

College of Medicine, University of the Philippines Manila, Pedro Gil Street, Malate, Manila, 1000, Philippines.

出版信息

Exp Parasitol. 2021 Feb;221:108049. doi: 10.1016/j.exppara.2020.108049. Epub 2020 Dec 8.

DOI:10.1016/j.exppara.2020.108049
PMID:33307097
Abstract

Globally, ascariasis ranks as the second leading intestinal helminth infection. However, progress in developing better control strategies, such as vaccines, remains slow-paced. This study aims to measure antibody production and parasite load in male BALB/c mice immunized with crude Ascaris suum intestinal tract homogenate. Thirty-two (32) mice were randomized into: (1) unvaccinated, uninfected (UU); (2) unvaccinated, infected (UI); (3) vaccinated, uninfected (VU); and (4) vaccinated, infected (VI) groups. A 100-μL vaccine containing 50 μg of homogenized A. suum intestines and Complete Freund's Adjuvant (1:1) were introduced intraperitoneally. Immunizations were done on days 0, 10, and 20. Oral gavage with 1000 embryonated eggs was done on day 30. Blood was obtained at day 40. To measure serum IgG levels, indirect ELISA was done. Microtiter plates were coated with 100 μg larval homogenate, and HRP-conjugated anti-mouse IgG was used as secondary antibody. Parasite load was measured in lung and liver tissues. Tukey's HSD of signal to cut-off ratios of absorbance readings obtained in indirect ELISA procedure for the 1:200 serum dilution showed statistically significant difference between the UU and VI (p = 0.026) as well as between UI and VI (p = 0.003) groups. No statistically significant difference in parasite load was observed in the lungs (p = 0.074), liver (p = 0.130), and both lungs and liver (p = 0.101). Immunization elicited a significant larva-directed IgG production. However, there is no significant difference in parasite loads in either lung or liver tissues across all treatment groups as the larval counts obtained from the study were very low and may not be indicative of the actual parasite load in mice.

摘要

在全球范围内,蛔虫病是排名第二的主要肠道蠕虫感染。然而,在开发更好的控制策略方面,如疫苗,进展仍然缓慢。本研究旨在测量雄性 BALB/c 小鼠用粗制猪蛔虫肠道匀浆免疫后抗体产生和寄生虫负荷。将 32 只小鼠随机分为:(1)未接种、未感染(UU);(2)未接种、感染(UI);(3)接种、未感染(VU)和(4)接种、感染(VI)组。用含有 50μg 猪蛔虫肠道匀浆和完全弗氏佐剂(1:1)的 100μL 疫苗进行腹腔内注射。免疫在第 0、10 和 20 天进行。第 30 天用 1000 个卵进行口服灌胃。第 40 天采集血液。通过间接 ELISA 测量血清 IgG 水平。将微滴定板用 100μg 幼虫匀浆包被,并使用 HRP 标记的抗小鼠 IgG 作为二抗。在肺和肝组织中测量寄生虫负荷。间接 ELISA 程序中 1:200 血清稀释的吸光度读数的信号与截止比值的 Tukey HSD 显示 UU 和 VI 组(p=0.026)以及 UI 和 VI 组(p=0.003)之间存在统计学显著差异。在肺(p=0.074)、肝(p=0.130)和肺和肝(p=0.101)中未观察到寄生虫负荷的统计学显著差异。免疫诱导了显著的幼虫定向 IgG 产生。然而,由于从研究中获得的幼虫计数非常低,可能无法指示小鼠中实际的寄生虫负荷,因此所有治疗组的肺或肝组织中的寄生虫负荷没有差异。

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