title: [Activation of NOD2 signalling pathway stimulates the function of human dendritic cells loaded with leukemia cell lysates].
abstract: The purpose of this study was to explore the effect of NOD2 signalling pathway activated by muramyl dipeptide (MDP) on the immunomodulation effect of human monocyte-derived dendritic cells (DC) loaded with leukemia cell lysates. Peripheral blood mononuclear cells (PBMNC) were isolated by density gradient centrifugation, These cells were cultured with three cytokines for 7 days to induce their maturation. On the 5th day, cells were loaded with leukemia cell HL-60 lysates. NOD2 expression was detected by RT-PCR and Western blot. The phenotype of DC were analyzed by flow cytometry, and ELISA was used to assay levels of IL-12 (p40) . The results showed that MDP could trigger NOD2 mRNA and protein expression in different groups of DC, especially in sensitized DC+MDP group, which was significantly higher than that in the DC+MDP group and sensitized DC without MDP stimulation, the difference was statistically significant (P < 0.05). Besides, the expression of surface molecules (HLA-DR, CD80, CD83, CD86, CD40) in the group of DC loaded with leukemia cell lysate and stimulated by MDP (sensitized DC+MDP) reached the highest level, followed by the group of DC loaded with leukemia cell lysate without MDP and DC only stimulated by MDP, non-treated DC were the lowest (P < 0.05). Similarly, compared with untreated unstimulated DC, after loading with HL-60 lysates or only stimulating with MDP, the secretion of IL-12p40 increased, but IL-12p40 level (573.86 ± 32.09 pg/ml) in DC+MDP group was higher than that in group of sensitized DC (365.03 ± 28.86 pg/ml) (P < 0.05), and it in sensitized DC+MDP group reached the highest (898.30 ± 61.08) pg/ml, compared to other groups (P < 0.05). It is concluded that MDP can significantly enhance the NOD2 mRNA and protein expression in sensitized DC, promote the expression of HLA-DR, synergistic costimulatory molecules and adhesion molecules of DC, at the same time, MDP can increase secretion of inflammatory factors IL-12p40. This study will provide a new ideas for DC application in leukemia immunotherapy.
authors.affiliation: Department of Hematology, The First Clinical Medical College of Three Gorges University, Yichang 443000, Hubei Province, China.
authors.forename: Dan-Lei
authors.initials: DL
authors.lastname: Han
authors.affiliation: Department of Hematology, The First Clinical Medical College of Three Gorges University, Yichang 443000, Hubei Province, China.
authors.forename: Hai-Yan
authors.initials: HY
authors.lastname: Wang
authors.affiliation: Department of Hematology, The First Clinical Medical College of Three Gorges University, Yichang 443000, Hubei Province, China. E-mail:gjm425@163.com.
authors.forename: Jing-Ming
authors.initials: JM
authors.lastname: Guo
authors.affiliation: Department of Hematology, The First Clinical Medical College of Three Gorges University, Yichang 443000, Hubei Province, China.
authors.forename: Hong
authors.initials: H
authors.lastname: Yi
authors.affiliation: Department of Hematology, The First Clinical Medical College of Three Gorges University, Yichang 443000, Hubei Province, China.
authors.forename: Yi-Qin
authors.initials: YQ
authors.lastname: Zeng
authors.affiliation: Department of Hematology, The First Clinical Medical College of Three Gorges University, Yichang 443000, Hubei Province, China.
authors.forename: Hong
authors.initials: H
authors.lastname: Ai
mesh_terms: D000119:Acetylmuramyl-Alanyl-Isoglutamine / Q000494:pharmacology; D002478:Cells, Cultured; D003713:Dendritic Cells / Q000276:immunology* / Q000378:metabolism*; D018922:HL-60 Cells; D006801:Humans; D053762:Interleukin-12 Subunit p40 / Q000378:metabolism; D007938:Leukemia / Q000276:immunology* / Q000378:metabolism; D007963:Leukocytes, Mononuclear / Q000378:metabolism; D008565:Membrane Proteins / Q000378:metabolism; D053473:Nod2 Signaling Adaptor Protein / Q000378:metabolism*
chemical_list: D053762:Interleukin-12 Subunit p40; D008565:Membrane Proteins; C429018:NOD2 protein, human; D053473:Nod2 Signaling Adaptor Protein; D000119:Acetylmuramyl-Alanyl-Isoglutamine