[Preliminary study on lymphotactin expression of monocyte-derived dendritic cells].
OBJECTIVE
To induce dendritic cells (DCs) from human peripheral blood monocytes in vitro and to investigate the kinetics lymphotactin (Lptn) mRNA expression.


METHODS
Monocytes were isolated from normal human peripheral blood cells by density gradient centrifugation and the plastic-adherent cells were cultured with the cytokines (rhGM-CSF, rhIL-4, rhTNF-alpha). The immature and mature DCs' surface antigens CD1a and CD83 were analyzed by flow cytometry (FCM). The morphology of mature DCs induced by rhTNF-alpha was observed under electron microscope. Lptn cDNA was amplified by RT-PCR, cloned to vector pGM-T Easy and completely sequenced. The RT-PCR products of cells were separated on gel and analyzed by semi-quantification.


RESULTS
The cells cultured for 7 d exhibited special dendritic morphology of mature DCs and highly expressed CD83. The sequence of cDNA was confirmed and was identical to that of NO. U23772 published in GenBank. The Lptn mRNA expression of DCs was stronger in DCs cultured for 7 d than in those for 5 d, whereas it was not detected in those for 3 d.


CONCLUSIONS
The monocyte-derived dendritic cells can express Lptn mRNA in a maturation-dependent manner.
Lu
Xiong-Bing
XB
Department of Urology, Second Affiliated Hospital, Nanchang University, Nanchang 330006, China. luxiongbing123456@yahoo.com.cn
Xie
Hui-Li
HL
Huang
Hong-Wei
HW
Shi
Zi-Min
ZM
Xiong
Jian-Hua
JH
Pan
Zheng-Yue
ZY
D000199:Actins / Q000235:genetics; D000818:Animals; D002478:Cells, Cultured; D003001:Cloning, Molecular; D003713:Dendritic Cells / Q000166:cytology* / Q000378:metabolism* / Q000648:ultrastructure; D004587:Electrophoresis, Agar Gel; D006801:Humans; D007700:Kinetics; D008222:Lymphokines / Q000235:genetics*; D051379:Mice; D046529:Microscopy, Electron, Transmission; D009000:Monocytes / Q000166:cytology*; D012333:RNA, Messenger / Q000235:genetics / Q000378:metabolism; D020133:Reverse Transcriptase Polymerase Chain Reaction; D012795:Sialoglycoproteins / Q000235:genetics*
D000199:Actins; D008222:Lymphokines; D012333:RNA, Messenger; D012795:Sialoglycoproteins; C018997:lymphotactin