Effect of in situ expression of human interleukin-6 on antibody responses against Salmonella typhimurium antigens
Effect of in situ expression of human interleukin-6 on antibody responses against Salmonella typhimurium antigens
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DOI：
10.1016/s0928-8244(03)00066-x
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发表时间：
2003-07-15
影响因子：
--
通讯作者：
Hahn, HP
中科院分区：
文献类型：
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作者：
Li, YY;Reichenstein, K;Hahn, HP
为了激发黏膜对细菌表面抗原的分泌性免疫应答，我们构建了一株分泌人白细胞介素6(HIL)-6的鼠伤寒沙门氏菌X4064(pCH1A+pYL3E)，利用溶血素(Hly)输出子分泌性递送功能性hIL-6-溶血素融合蛋白(hIL-6-hlyA(S))。通过将第二个hIL-6-hlyA(S)表达载体(PYL3E)稳定地导入X4064(PCH1A)菌株中，hIL-6-hlyA(S)的分泌效率至少提高了10倍。与亲本菌株中的pCH1a一样，pYL3E在没有抗生素选择的情况下在体外是稳定的，在体内也没有质粒对其稳定性的干扰。增加hIL-6-hlyA(S)的表达不会对细菌生长产生不利影响。小鼠口服不同hIL-6分泌株的比较免疫实验表明，原位hIL-6的产生增加影响了系统对沙门氏菌抗原的抗体应答，但对粘膜应答无明显影响。在X4064(pCH1A+pYL3E)免疫的小鼠中，与X4064(PCH1A)免疫的小鼠和hIL-6阴性对照株相比，小鼠血清中脂多糖(LPS)的IgG和IgA滴度显著升高。较高的血清抗体滴度伴随着脾和Peyer‘s斑块中免疫球蛋白和免疫球蛋白A特异性抗体分泌细胞的增加。这些数据表明，针对沙门氏菌内毒素的系统抗体反应在很大程度上受IL-6的影响，此外，重组表达的IL-6的数量和细胞位置似乎对增强免疫反应至关重要。(C)2003年欧洲微生物学会联合会。爱思唯尔科学公司出版。版权所有。
In an attempt to trigger increased mucosal secretory immune responses against bacterial surface antigens, we constructed an optimized human interleukin (hIL)-6-secreting Salmonella typhimurium strain (X4064(pCH1A+pYL3E)), utilizing the hemolysin (Hly) exporter for secretory delivery of a functional hIL-6-hemolysin fusion protein (hIL-6-HlyA(s)). Through stable introduction of a second hIL-6-HlyA(s) expression plasmid (pYL3E) in the previously described X4064(pCH1A) strain, hIL-6-HlyA(s) secretion efficiencies were increased by at least 10-fold. As pCH1A in the parental strain, pYL3E was stable in vitro in the absence of antibiotic selection and in vivo neither did plasmids interfere in their stabilities. Increased hIL-6-HlyA(s) expression did not adversely interfere with bacterial growth. Comparative immunization experiments in mice with oral application of the different hIL-6-secreting strains revealed that increased in situ hIL-6-production influenced systemic antibody responses against Salmonella antigens but had no marked effect on mucosal responses. In mice immunized with X4064(pCH1A+pYL3E) significantly higher sera IgG and IgA titers for lipopolysaccharide (LPS) were found compared to mice immunized with X4064(pCH1A) and a hIL-6-negative control strain. Higher sera antibody titers were accompanied by increased numbers of IgG- and IgA-specific antibody-secreting cells in spleens and Peyer's patches, respectively. These data suggest that systemic antibody responses against Salmonella LPS are largely effected by IL-6 and, moreover, the amount and the cellular location of recombinantly expressed IL-6 appears to be crucial for enhancement of immune responses. (C) 2003 Federation of European Microbiological Societies. Published by Elsevier Science B.V. All rights reserved.