[Expression of cytokines in murine BMDCs induced by recombinant E.coli LLO/OVA via TLR4 and NOD1 receptors].
OBJECTIVE
To investigate the relationship between the activation of pattern recognition receptors and the cytokines expression of dendritic cells.


METHODS
After bone marrow-derived dendritic cells (BMDCs) were pulsed by E.coli LLO/OVA, the mRNA expression of pattern recognition receptors and the downstream NF-kappaB signal pathway associated molecules were detected by microarray hybridization and RT-PCR; the expression of costimulatory molecules, MHC class II and cytokines were determined by flow cytometry and ELISA.


RESULTS
After BMDCs were pulsed by E.coli LLO/OVA for 2 h, the levels of TLR4, Myd88, Rip2, Irak1, Irak2, Ikkalpha, NF-kappaB1 and NF-kappaB2 mRNA up-regulated; and the levels of Icam1, IL-1a, IL-1b, IL-6 and TNF-alpha mRNA up-regulated also; after BMDCs were pulsed by E.coli LLO/OVA for 4 h, the levels of Card4(Nod1), Rip2, Ikkbeta, NF-kappaB1 and NF-kappaB2 mRNA up-regulated, meanwhile, the levels of TNF-gamma, TNF-beta and CD40 mRNA up-regulated. The expressions of costimulatory molecules and MHC class II of the BMDCs up-regulated and the concentration of IL-12 and IFN-gamma increased in the supernatant of BMDCs pulsed by E.coli LLO/OVA at 24 h.


CONCLUSIONS
Recombinant E.coli LLO/OVA induces maturation and expression of cytokines, especially IL-12 and IFN-gamma, of murine BMDCs through activation of NF-kappaB signal pathway with TLR4 and NOD1 receptors.
Xu
Man
M
Department of Pathology, Chongqing University of Medical Sciences, China. manxucqmu@yahoo.com
Dai
Ming-Shen
MS
Mi
Can
C
D000818:Animals; D001427:Bacterial Toxins / Q000235:genetics / Q000378:metabolism / Q000494:pharmacology*; D001854:Bone Marrow Cells / Q000166:cytology*; D016207:Cytokines / Q000235:genetics*; D003713:Dendritic Cells / Q000166:cytology / Q000187:drug effects / Q000378:metabolism*; D004797:Enzyme-Linked Immunosorbent Assay; D004926:Escherichia coli / Q000235:genetics / Q000378:metabolism; D005434:Flow Cytometry; D006360:Heat-Shock Proteins / Q000235:genetics / Q000378:metabolism / Q000494:pharmacology*; D006460:Hemolysin Proteins / Q000235:genetics / Q000378:metabolism / Q000494:pharmacology*; D018799:Intercellular Adhesion Molecule-1 / Q000235:genetics; D053592:Interleukin-1 Receptor-Associated Kinases / Q000235:genetics; D015850:Interleukin-6 / Q000235:genetics; D008233:Lymphotoxin-alpha / Q000235:genetics; D051379:Mice; D053594:Myeloid Differentiation Factor 88 / Q000235:genetics; D052002:NF-kappa B p50 Subunit / Q000235:genetics; D052003:NF-kappa B p52 Subunit / Q000235:genetics; D053474:Nod1 Signaling Adaptor Protein / Q000235:genetics / Q000502:physiology*; D020411:Oligonucleotide Array Sequence Analysis; D010047:Ovalbumin / Q000235:genetics / Q000378:metabolism / Q000494:pharmacology*; D020133:Reverse Transcriptase Polymerase Chain Reaction; D051197:Toll-Like Receptor 4 / Q000235:genetics / Q000502:physiology*; D014409:Tumor Necrosis Factor-alpha / Q000235:genetics
D001427:Bacterial Toxins; D016207:Cytokines; D006360:Heat-Shock Proteins; D006460:Hemolysin Proteins; C090509:Icam1 protein, mouse; D015850:Interleukin-6; D008233:Lymphotoxin-alpha; D053594:Myeloid Differentiation Factor 88; D052002:NF-kappa B p50 Subunit; D052003:NF-kappa B p52 Subunit; D053474:Nod1 Signaling Adaptor Protein; D051197:Toll-Like Receptor 4; D014409:Tumor Necrosis Factor-alpha; D018799:Intercellular Adhesion Molecule-1; D010047:Ovalbumin; D053592:Interleukin-1 Receptor-Associated Kinases; C507769:Irak1 protein, mouse; C038705:hlyA protein, Listeria monocytogenes