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


CONCLUSIONS
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.
Department of Pathology, Molecular Medicine and Cancer Research Centre, Chongqing Medical University, Chongqing 400016, China. manxucqmu08@yahoo.cn
Man
M
Xu
Ming-Shen
MS
Dai
D000818:Animals; D000951:Antigens, Neoplasm / Q000235:genetics / Q000494:pharmacology; D001427:Bacterial Toxins / Q000235:genetics / Q000494:pharmacology*; D019496:Cancer Vaccines / Q000235:genetics / Q000494:pharmacology*; D045744:Cell Line, Tumor; D049109:Cell Proliferation / Q000187:drug effects; D002470:Cell Survival / Q000187:drug effects / Q000276:immunology; D018920:Coculture Techniques; D016207:Cytokines / Q000276:immunology / Q000378:metabolism; D003713:Dendritic Cells / Q000166:cytology / Q000187:drug effects* / Q000276:immunology / Q000378:metabolism; D004797:Enzyme-Linked Immunosorbent Assay; D004926:Escherichia coli / Q000235:genetics / Q000378:metabolism; D005260:Female; D005434:Flow Cytometry; D006360:Heat-Shock Proteins / Q000235:genetics / Q000494:pharmacology*; D006460:Hemolysin Proteins / Q000235:genetics / Q000494:pharmacology*; D007113:Immunity, Innate / Q000187:drug effects*; D051379:Mice; D008810:Mice, Inbred C57BL; D053474:Nod1 Signaling Adaptor Protein / Q000235:genetics / Q000502:physiology*; D010047:Ovalbumin / Q000235:genetics / Q000494:pharmacology*; D011993:Recombinant Fusion Proteins / Q000235:genetics / Q000494:pharmacology*; D020133:Reverse Transcriptase Polymerase Chain Reaction; D013602:T-Lymphocytes, Cytotoxic / Q000187:drug effects / Q000276:immunology*; D051197:Toll-Like Receptor 4 / Q000235:genetics / Q000502:physiology*
D000951:Antigens, Neoplasm; D001427:Bacterial Toxins; D019496:Cancer Vaccines; D016207:Cytokines; D006360:Heat-Shock Proteins; D006460:Hemolysin Proteins; D053474:Nod1 Signaling Adaptor Protein; D011993:Recombinant Fusion Proteins; D051197:Toll-Like Receptor 4; D010047:Ovalbumin; C038705:hlyA protein, Listeria monocytogenes