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用表达轮状病毒 VP8* 蛋白的乳球菌对小鼠进行口服免疫。

Oral immunization of mice with Lactococcus lactis expressing the rotavirus VP8* protein.

机构信息

Departamento de Microbiología, Facultad de Medicina, Universitat de València, Valencia, Spain.

出版信息

Biotechnol Lett. 2011 Jun;33(6):1169-75. doi: 10.1007/s10529-011-0551-6. Epub 2011 Feb 8.

Abstract

The efficacy of recombinant Lactococcus lactis as a delivery vehicle for a rotavirus antigen was evaluated in a mouse model. The rotavirus VP8* protein was expressed intracellularly and extracellularly in L. lactis wild type and in an alr mutant deficient in alanine racemase activity, necessary for the synthesis of the cell-wall component D: -alanine. When the mucosal immune response was evaluated by measuring VP8*-specific IgA antibody in faeces, wild-type L. lactis triggered a low IgA synthesis only when the secreting strain was used. In contrast, VP8*-specific IgA was detected in faeces of both groups of mice orally given the alr mutant expressing extracellular VP8* and intracellular VP8*, which reached levels similar to that obtained with the wild type secreting strain. However, oral administration of the recombinant strains did not induce serum IgG or IgA responses. L. lactis cell-wall mutants may therefore provide certain advantages when low-antigenic proteins are expressed intracellularly. However, the low immune response obtained by using this antigen-bacterial host combination prompts to the use of new strains and vaccination protocols in order to develop acceptable rotavirus immunization levels.

摘要

乳球菌作为轮状病毒抗原传递载体的功效在小鼠模型中进行了评估。轮状病毒 VP8蛋白在乳球菌野生型和丙氨酸消旋酶活性缺陷的 alr 突变体中进行了细胞内和细胞外表达,丙氨酸消旋酶活性是细胞壁成分 D: -丙氨酸合成所必需的。通过测量粪便中的 VP8-特异性 IgA 抗体来评估黏膜免疫反应时,只有使用分泌型菌株时,野生型乳球菌才会引发低水平的 IgA 合成。相比之下,经口给予表达细胞外 VP8和细胞内 VP8的 alr 突变体的两组小鼠粪便中均检测到 VP8*-特异性 IgA,其水平与使用分泌型野生型菌株获得的水平相似。然而,重组菌株的口服给药并未诱导血清 IgG 或 IgA 反应。因此,当表达低抗原性蛋白时,乳球菌细胞壁突变体可能具有某些优势。然而,使用这种抗原-细菌宿主组合获得的低免疫反应促使使用新的菌株和接种方案,以开发可接受的轮状病毒免疫水平。

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