重组大肠杆菌 LLO/OVA 通过 TLR4 和 NOD1 受体诱导小鼠 BMDCs 成熟，并促进特异性细胞毒性 T 细胞免疫。

Recombinant E. coli LLO/OVA induces murine BMDCs maturation via TLR4 and NOD1 receptor and promotes specific cytotoxic T cell immunity.

机构信息

Department of Pathology, Molecular Medicine and Cancer Research Centre, Chongqing Medical University, Chongqing 400016, China.

出版信息

Biomed Environ Sci. 2010 Oct;23(5):350-6. doi: 10.1016/S0895-3988(10)60075-X.

DOI:10.1016/S0895-3988(10)60075-X

PMID:21112482

Abstract

OBJECTIVE

To explore the immune stimulation effect of recombinant E.coli LLO/OVA on mice bone marrow-derived dendritic cells (BMDCs) and T lymphocytes in vitro.

METHODS

After BMDCs stimulated by E.coli LLO/OVA, their Toll-like receptor (TLR) and nucleotide-binding oligomerization domain (NOD) receptor signalling pathway were examined by superarray hybridization; and the priming effect of the vaccine activated BMDCs on CD4(+)T and CD8(+)T was determined by [3H]thymidine uptake and ELISA, the tumor cytotoxic effect of activated CD8(+)T cells was determined by cytotoxic assay.

RESULTS

After BMDCs were activated by E. coli LLO/OVA via TLR4, NOD1 receptor and NF-κB signalling pathway, the expression of their surface molecules including MHC class I, MHC class II, CD40, CD80 and CD86 significantly up-regulated; the secretion of IL-12 and IFN-γ increased also. The mature BMDCs stimulated the allergic CD4(+)T and CD8(+)T cells proliferation and their IL-2 and IFN-γ secretion, and the activated CD8(+)T cells effectively killed B16-OVA melanoma cells and RMA-S/OVA lymphoma cells in vitro.

CONCLUSION

E.coli LLO/OVA is effective in inducing BMDCs maturation via activating TLR4 and NOD1 receptor signalling pathway and promoting specific anti-tumor T cell immunity in vitro.

摘要

目的

探讨重组大肠埃希菌 LLO/OVA 对小鼠骨髓来源树突状细胞（BMDC）和 T 淋巴细胞的体外免疫刺激作用。

方法

用大肠埃希菌 LLO/OVA 刺激 BMDC 后，通过超级杂交检测其 Toll 样受体（TLR）和核苷酸结合寡聚化结构域（NOD）受体信号通路；通过[3H]胸苷摄取和 ELISA 检测疫苗激活的 BMDC 对 CD4(+)T 和 CD8(+)T 的启动作用，通过细胞毒性测定检测激活的 CD8(+)T 细胞的肿瘤细胞毒性作用。

结果

大肠埃希菌 LLO/OVA 通过 TLR4、NOD1 受体和 NF-κB 信号通路激活 BMDC 后，其表面分子如 MHC Ⅰ类、MHC Ⅱ类、CD40、CD80 和 CD86 的表达明显上调；IL-12 和 IFN-γ 的分泌也增加。成熟的 BMDC 刺激过敏 CD4(+)T 和 CD8(+)T 细胞增殖及其 IL-2 和 IFN-γ 的分泌，激活的 CD8(+)T 细胞有效杀伤体外 B16-OVA 黑素瘤细胞和 RMA-S/OVA 淋巴瘤细胞。