[Acceleration of mixed lymphocyte reaction by HCV C-Fc gene-transferred murine dendritic cells].
OBJECTIVE
To observe the metergasis of murine dendritic cells (DCs) transfected with HCV C-Fc gene through electroporation.


METHODS
Mononucleocytes isolated from murine bone marrow were co-cultured with rmGM-CSF and rm-IL-4 for 7 days. Morphological characteristics of the cultured cells were observed under scan electron-microscope (SEM) and the expression of DEC205 on the cells was detected by FACS. DCs derived from the culture were transfected with plasmids containing HCV C-Fc gene. HCV C-Fc level in the transfected cells was detected by indirect immunofluorescence assay. MLR was studied with DCs and T cells.


RESULTS
Following 7-day culture, a large number of cells with typical characteristics of DC were observed. The HCV C-Fc level in the transfected DCs was higher. MLR was stimulated markedly by DCs transfected with HCV C-Fc gene in comparison with the control group.


CONCLUSIONS
A large number of DCs could be generated from murine bone marrow mononucleocyte cultures supplemented with GM-CSF and IL-4 for 1 week. The function of DCs transfected with pcDNA3HCV C-Fc was enhanced in MLR.
Wang
Quan-chu
QC
The Center of Diagnosis and Treatment of Infection Diseases of PLA, Tangdu Hospital, Fourth Military Medical University, Xi'an 710038, China. quanchuwang998@hotmail.com
Feng
Zhi-hua
ZH
Zhou
Yong-xing
YX
Nie
Qing-he
QH
Bai
Xue-fan
XF
D000818:Animals; D019677:Artificial Gene Fusion; D002478:Cells, Cultured; D018920:Coculture Techniques; D003713:Dendritic Cells / Q000378:metabolism*; D005260:Female; D016174:Hepacivirus / Q000235:genetics*; D018936:Hepatitis C Antigens / Q000096:biosynthesis / Q000235:genetics; D007141:Immunoglobulin Fc Fragments / Q000096:biosynthesis / Q000235:genetics*; D007074:Immunoglobulin G / Q000096:biosynthesis / Q000235:genetics; D007959:Lymphocyte Culture Test, Mixed; D051379:Mice; D008807:Mice, Inbred BALB C; D011993:Recombinant Fusion Proteins / Q000096:biosynthesis / Q000235:genetics; D013601:T-Lymphocytes / Q000166:cytology; D014162:Transfection; D014758:Viral Core Proteins / Q000096:biosynthesis / Q000235:genetics*
D018936:Hepatitis C Antigens; D007141:Immunoglobulin Fc Fragments; D007074:Immunoglobulin G; D011993:Recombinant Fusion Proteins; D014758:Viral Core Proteins; C433031:hepatitis C protein F, Hepatitis C virus