title: Construction and evaluation of rats' tolerogenic dendritic cells (DC) induced by NF-κB Decoy method.
abstract: OBJECTIVE
  To construct and evaluate rats' tolerogenic dendritic cells (DC) through induction by NF-κB Decoy method.
  
  
  METHODS
  GM-CSF and IL-4 were used to transform rats's monocytes into DC, and DC were stimulated with LPS, NF-κB Decoy ODN, and loaded with Bovine Type II Collagen. The following methods were employed to phenotype DC: 1) Observation of cell morphology; 2) Evaluation of cell viability using trypan blue staining; 3) Purity determination of DC through detection of specific markers OX-62; 4) Evaluation of mature state of DC via the determination of the expression of CD80 and CD86; 5) Determination of stimulation capability towards the proliferation of lymphocyte and the secretion of INF-r and IL-10.
  
  
  RESULTS
  The activity of DC was more than 92%, and the expression of OX-62 was more than 70%. Most of DC exhibited the phenotype of CD80(+)/CD86(-). Compared with control group and LPS-stimulation group, the less mature adhered cells and hairlike DC were observed in NF-κB decoy group. Significant reduction (p < 0.05) was observed for the positive expression and extension of CD80 and CD86 in cell surface. After loaded with calf type II collagen, the low expression of CD80 and CD86 remains to be existed. The stimulation capability of DC towards lymphocyte in NF-κB decoy group was lower than that in control group (p<0.05) and LPS stimulation group (p < 0.05).
  
  
  CONCLUSIONS
  NF-κB Decoy ODN method can be successfully applied for construct rats' tolerogenic dendritic cells (DC) with stable morphology and phenotype. The tolerogenic DC exhibited immature immune phenotype, and low capability to stimulate lymphocytes.
authors.affiliation: Department of Microbiology and Immunology, Affiliated Hospital of Guiyang Medical College, Guiyang Medical College, Guiyang, Guizhou, 550004, China.
authors.forename: HongMei
authors.initials: H
authors.lastname: Jiang
authors.affiliation: Department of Clinical Laboratory Hematology, Affiliated Hospital of Guiyang Medical College, Guiyang Medical College, Guiyang, Guizhou, 550004, China.
authors.forename: YaLi
authors.initials: Y
authors.lastname: Zhang
authors.affiliation: Department of Clinical Laboratory, Xiamen Second People's Hospital, Xiamen, Fujian, 361021, China.
authors.forename: XiangFei
authors.initials: X
authors.lastname: Yin
authors.affiliation: Department of Clinical Laboratory, The Third Hospital Subsidiary of Bengbu Medical College, Suzhou, Anhui, 34000, China.
authors.forename: HengGui
authors.initials: H
authors.lastname: Hu
authors.affiliation: Department of Clinical Laboratory, the Central Hospital of Lishui, Zhejiang 323000, China.
authors.forename: XiaoLei
authors.initials: X
authors.lastname: Hu
authors.affiliation: Department of Microbiology and Immunology, Affiliated Hospital of Guiyang Medical College, Guiyang Medical College, Guiyang, Guizhou, 550004, China.
authors.forename: Ying
authors.initials: Y
authors.lastname: Fei
authors.affiliation: Department of Experimental Surgery, Tangdu Hospital, Fourth Military Medical University, Xi'an 710038 China.
authors.forename: Yanyang
authors.initials: Y
authors.lastname: Tu
authors.affiliation: Department of Experimental Surgery, Tangdu Hospital, Fourth Military Medical University, Xi'an 710038 China.
authors.forename: Yongsheng
authors.initials: Y
authors.lastname: Zhang
mesh_terms: D000818:Animals; D015703:Antigens, CD; D000943:Antigens, Differentiation; D018122:B7-1 Antigen / Q000096:biosynthesis / Q000235:genetics* / Q000276:immunology; D051940:B7-2 Antigen / Q000096:biosynthesis / Q000235:genetics* / Q000276:immunology; D002465:Cell Movement / Q000187:drug effects / Q000276:immunology*; D003713:Dendritic Cells / Q000166:cytology / Q000187:drug effects / Q000276:immunology* / Q000378:metabolism*; D005786:Gene Expression Regulation / Q000187:drug effects; D016130:Immunophenotyping; D015847:Interleukin-4; D008297:Male; D016328:NF-kappa B / Q000235:genetics* / Q000276:immunology*; D009838:Oligodeoxyribonucleotides / Q000276:immunology / Q000494:pharmacology*; D009930:Organic Chemicals; D051381:Rats
chemical_list: D015703:Antigens, CD; D000943:Antigens, Differentiation; D018122:B7-1 Antigen; D051940:B7-2 Antigen; D016328:NF-kappa B; C468119:NF-kappaB decoy; C459465:OX-62 antigen, rat; D009838:Oligodeoxyribonucleotides; D009930:Organic Chemicals; C029248:Tolerogen; D015847:Interleukin-4
keywords: NF-κB Decoy ODN; calf type II collagen; dendritic cells (DC)