[Establishment of cell line transfected with human BTLA gene and preliminary study on its biological function].
OBJECTIVE
To establish 293T cells expressing human BTLA gene, a novel attenuator for B and T lymphocyte.


METHODS
The gene encoding human BTLA was amplified by RT-PCR from PHA activated T cells, and then the target gene fragment was inserted into retrovirus vector pEGZ-Term after being digested with EcoR I and BamH I. The recombinant vector was transfected into 293T cells with LipfectAMINE 2000, and the cells was further selected with Zeocin. Effect of 293T/BTLA cells on T cells proliferation and activation in vitro was been studied by methods of MTT and cytometry.


RESULTS
The stable expression of human BTLA on the transfected cell line was identified by flow cytometry analysis. In vitro, 293T/BTLA cells had an inhibitory effect on the proliferation of T cells stimulated by anti-CD3 mAb and down-regulated the expression of activation marker CD25 on the surface of T cells and decreased the secretion of IFN-gamma and IL-10.


CONCLUSIONS
A cell line 293T/BTLA stably expressing the human BTLA protein has been obtained and it can partially inhibit the proliferation and activation of T cells in vitro.
Wang
Xue-Feng
XF
Institute of Medical Biotechnology, Soochow University, Key Laboratory of Stem Cell of Jiangsu Province, Suzhou, China. wangxuefengsuda@hotmail.com
Chen
Yong-Jing
YJ
Wang
Qin
Q
Yang
Ke-Fang
KF
Dai
Qun
Q
Wang
Feng-Ming
FM
Zhang
Xue-Guang
XG
D000911:Antibodies, Monoclonal / Q000494:pharmacology; D017252:CD3 Complex / Q000276:immunology; D002460:Cell Line; D049109:Cell Proliferation / Q000187:drug effects; D004797:Enzyme-Linked Immunosorbent Assay; D005434:Flow Cytometry; D005822:Genetic Vectors / Q000235:genetics; D006801:Humans; D007371:Interferon-gamma / Q000378:metabolism; D016753:Interleukin-10 / Q000378:metabolism; D008213:Lymphocyte Activation / Q000235:genetics; D017463:Receptors, Complement 3b / Q000378:metabolism; D011971:Receptors, Immunologic / Q000235:genetics / Q000502:physiology*; D020133:Reverse Transcriptase Polymerase Chain Reaction; D013601:T-Lymphocytes / Q000166:cytology / Q000378:metabolism; D014162:Transfection
D000911:Antibodies, Monoclonal; C482586:BTLA protein, human; D017252:CD3 Complex; D017463:Receptors, Complement 3b; D011971:Receptors, Immunologic; D016753:Interleukin-10; D007371:Interferon-gamma