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疫苗研发:口服源自49.8 kDa菌毛蛋白亚基的肽可激活小鼠的肠道免疫反应。

vaccine development: Oral administration of peptides derived from the 49.8 kDa pili protein subunit activates the intestinal immune response in mice.

作者信息

Anam Khoirul, Endharti Agustina Tri, Poeranto Sri, Sujuti Hidayat, Hidayati Dwi Yuni Nur, Prawiro Sumarno Reto

机构信息

Doctoral Program in Medical Science, Faculty of Medicine, Universitas Brawijaya, Malang, Indonesia.

Study Program of Medical Laboratory Technology, Institute of Health and Science Technology Wiyata Husada, Samarinda, Indonesia.

出版信息

Vet World. 2022 Feb;15(2):281-287. doi: 10.14202/vetworld.2022.281-287. Epub 2022 Feb 11.

Abstract

BACKGROUND AND AIM

The morbidity and mortality of infections remain a global challenge. Epitope-based vaccine development is an emerging strategy to prevent bacterial invasion. This study aimed to identify the ability of the 49.8 kDa pili subunit adhesin protein epitope of to induce an intestinal immune response in mice.

MATERIALS AND METHODS

Thirty adult male Balb/c mice were divided into a control group, cholera toxin B subunit (CTB) group, CTB+QSSTGTNSQSDLDS (pep_1) group, CTB+DTTITKAETKTVTKNQVVDTPVTTDAAK (pep_2) group, and CTB+ ATLGATLNRLDFNVNNK (pep_3). We performed immunization by orally administering 50 μg of antigen and 50 μl of adjuvant once a week over 4 weeks. We assessed the cellular immune response by quantifying T helper 2 (Th2) and Th17 using flow cytometry. In addition, we assessed the humoral immune response by quantifying interleukin (IL-4), IL-17, secretory immunoglobulin A (sIgA), and β-defensin using enzyme-linked immunoassay. Statistical analysis was performed using one-way analysis of variance and Kruskal-Wallis test.

RESULTS

Peptide oral immunization increases the cellular immune response as reflected by the increase of Th2 (p=0.019) and Th17 (p=0.004) cell counts, particularly in the CTB_pep_1 group. Humoral immune response activation was demonstrated by increased IL-4 levels, especially in the CTB+pep_3 group (p=0.000). The IL-17 level was increased significantly in the CTB+pep_1 group (p=0.042). The mucosal immune response was demonstrated by the sIgA levels increase in the CTB+pep_3 group (p=0.042) and the β-defensin protein levels (p=0.000).

CONCLUSION

All selected peptides activated the cellular and humoral immune responses in the intestine of mice. Further studies are necessary to optimize antigen delivery and evaluate whether the neutralizing properties of these peptides allow them to prevent bacterial infection.

摘要

背景与目的

感染的发病率和死亡率仍是一项全球性挑战。基于表位的疫苗开发是预防细菌入侵的一种新兴策略。本研究旨在确定49.8 kDa菌毛亚基粘附素蛋白表位诱导小鼠肠道免疫反应的能力。

材料与方法

将30只成年雄性Balb/c小鼠分为对照组、霍乱毒素B亚基(CTB)组、CTB+QSSTGTNSQSDLDS(肽_1)组、CTB+DTTITKAETKTVTKNQVVDTPVTTDAAK(肽_2)组和CTB+ATLGATLNRLDFNVNNK(肽_3)组。我们通过每周口服一次50 μg抗原和50 μl佐剂进行免疫,共持续4周。我们使用流式细胞术通过定量辅助性T细胞2(Th2)和Th17来评估细胞免疫反应。此外,我们使用酶联免疫吸附测定法通过定量白细胞介素(IL-4)、IL-17、分泌型免疫球蛋白A(sIgA)和β-防御素来评估体液免疫反应。使用单因素方差分析和克鲁斯卡尔-沃利斯检验进行统计分析。

结果

肽口服免疫增加了细胞免疫反应,这通过Th2(p=0.019)和Th17(p=0.004)细胞计数的增加得以体现,特别是在CTB_肽_1组。体液免疫反应的激活通过IL-4水平的升高得以证明,尤其是在CTB+肽_3组(p=0.000)。CTB+肽_1组中IL-17水平显著升高(p=0.042)。CTB+肽_3组中sIgA水平的增加(p=0.042)和β-防御素蛋白水平(p=0.000)证明了黏膜免疫反应。

结论

所有选定的肽均激活了小鼠肠道中的细胞免疫和体液免疫反应。有必要进一步研究以优化抗原递送,并评估这些肽的中和特性是否使其能够预防细菌感染。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/8714/8980390/47656e6b434a/Vetworld-15-281-g001.jpg

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