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鼻内接种丝氨酸/苏氨酸磷酸酶 2A 合成肽和重组抗原对安氏旋尾线虫的有效性。

Effectiveness of intranasal vaccination against Angiostrongylus costaricensis using a serine/threonine phosphatase 2 A synthetic peptide and recombinant antigens.

机构信息

Institute of Biotechnology, Biochemistry and Molecular Parasitology Group, University of Granada, Edif Mecenas, Campus Fuentenueva, 18071 Granada, Spain.

出版信息

Vaccine. 2010 Jul 19;28(32):5185-96. doi: 10.1016/j.vaccine.2010.05.072. Epub 2010 Jun 15.

Abstract

Intranasal immunization was assayed in C57BL/6 mice against Angiostrongylus costaricensis using a synthetic and a recombinant peptide belonging to the catalytic region of the serine/threonine phosphatase 2 A (PP2A) of the parasite. Immunization was carried out with the synthetic peptide (SP) polymerized either with itself or with the beta fraction of the cholera toxin (CTB) and then enclosed in nanocapsules of phosphatidyl choline, cholesterol and Quil A (ISCOM). Another group of mice was immunized with recombinant peptide. Immunization consisted of two intranasal inoculations at two-week intervals, and the challenge with L3 larvae was made one month after the last vaccination. The effectiveness of immunization was evaluated 30 days after infection by analysis of the number of parasites in the arteries of the immunized mice, as well as by measuring spleen sizes in the experimental groups. The response induced was determined by identifying the isotypes of IgG as well as the IgE and IgA specific antigen response. The interleukins produced by the splenocyte culture of the different groups were assessed after exposing them to the peptide used in the immunization. From our results, 60%, 80%, and 100% protection against the A. costaricensis challenge was achieved in mice immunized with polymerized synthetic peptide in ISCOM, synthetic peptide polymerized with the CTB in ISCOM and inclusion bodies respectively. Splenomegaly was found to be less evident in the immunized mice than in the controls. A significant increase in IFN gamma and IL-17 levels was observed in the group with 100% protection. The results showed that vaccination through the nasal mucosa may constitute a useful method of immunization and result in a protective immune response against A. costaricensis.

摘要

经鼻免疫接种在 C57BL/6 小鼠中针对 Angiostrongylus costaricensis 使用寄生虫丝氨酸/苏氨酸磷酸酶 2A(PP2A)的催化区域的合成肽和重组肽进行了检测。免疫接种是用合成肽(SP)自身聚合或与霍乱毒素(CTB)的β部分聚合,然后用磷脂酰胆碱、胆固醇和 Quil A(ISCOM)包裹纳米胶囊进行的。另一组小鼠用重组肽免疫。免疫接种包括两次两周间隔的经鼻接种,最后一次接种一个月后进行 L3 幼虫攻击。感染后 30 天通过分析免疫小鼠动脉中的寄生虫数量以及测量实验组的脾脏大小来评估免疫的有效性。通过鉴定 IgG 同种型以及 IgE 和 IgA 特异性抗原反应来确定诱导的反应。在用免疫中使用的肽暴露后,评估不同组的脾细胞培养物产生的白细胞介素。从我们的结果来看,用 ISCOM 中的聚合合成肽、ISCOM 中的 CTB 聚合合成肽和包涵体分别免疫的小鼠中,有 60%、80%和 100%的 A. costaricensis 攻击得到了保护。与对照组相比,免疫小鼠的脾肿大程度较轻。在 100%保护的组中观察到 IFN-γ和 IL-17 水平显著增加。结果表明,通过鼻腔黏膜进行疫苗接种可能构成一种有用的免疫方法,并导致针对 A. costaricensis 的保护性免疫反应。

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