[The change of immunoactivity of dendritic cells induced by recombinant rat 4-1BBL].
OBJECTIVE
To investigate the change of immunoactivity and surface marker of dendritic cells (DCs) derived from bone marrow and induced by rat 4-1BBL.


METHODS
Plasmid pIRES2-EGFP-4-1BBL was constructed and tansfected into HepG2 cells by lipofectin-mediated method, and positive cells were screened by G418. The expression of 4-1BBL in cells were detected by RT-PCR, and the expression of EGFP was detected by fluorescence microscopy. Various HepG2 cells were co-cultured with DC for 2 days, and then the expression of MHC-II and CD80 on DC were detected by FCM, and IL-6 and IL-12 levels secreted by DCs were tested by ELISA.


RESULTS
The recombinant pIRES2-EGFP-4-1BBL vector was successfully constructed and was steadily expressed in HepG2 cells transfected by pIRES2-EGFP-4-1BBL. Compared with HepG2 cells transfected by pIRES2-EGFP plasmid , recombinant HepG2 cells induced high expression of MHC-II and CD80 and secreted more IL-6 and IL-12 on DCs(P<0.05).


CONCLUSIONS
The recombinant rat 4-1BBL could promote the maturity of DCs derived from bone marrow.
Tan
Jian-Xin
JX
Department of Gastroenterology in First Hospital, Suzhou 215006, China. yjx13906210479@sina.com
Chen
Wei-Chang
WC
Zhang
Xue-Guang
XG
Zhang
Guang-Bo
GB
D053330:4-1BB Ligand / Q000235:genetics / Q000378:metabolism / Q000494:pharmacology*; D000818:Animals; D018122:B7-1 Antigen / Q000378:metabolism; D045744:Cell Line, Tumor; D003713:Dendritic Cells / Q000187:drug effects* / Q000276:immunology*; D004797:Enzyme-Linked Immunosorbent Assay; D005260:Female; D005802:Genes, MHC Class II; D006801:Humans; D018664:Interleukin-12 / Q000378:metabolism; D015850:Interleukin-6 / Q000378:metabolism; D008297:Male; D008856:Microscopy, Fluorescence; D051381:Rats; D017207:Rats, Sprague-Dawley; D011994:Recombinant Proteins / Q000235:genetics / Q000276:immunology* / Q000378:metabolism; D020133:Reverse Transcriptase Polymerase Chain Reaction; D014162:Transfection
D053330:4-1BB Ligand; D018122:B7-1 Antigen; D015850:Interleukin-6; D011994:Recombinant Proteins; D018664:Interleukin-12