[The isolation and expansion of gamma delta T cells in peripheral blood and bronchoalveolar lavage fluid].
OBJECTIVE
To explore the method of selective culture of gamma delta T cells in peripheral blood and bronchoalveolar lavage fluid(BALF) for obtaining pure gamma delta T cell subset.


METHODS
A novel attack-panning method for selectively culturing gamma delta T cells was set up. The peripheral blood mononuclear cells (PBMCs) and BALF were isolated by Ficoll-hypaque density gradient centrifugation (n=10). alpha beta T cells in PBMCs were depleted by complement-dependent-cytotoxicity(CDC) after the monocytes/macrophages were removed by adherence. The gamma delta T cells in PBMCs were cultivated selectively using anti-TCR gamma delta monoclonal antibody and IL-2. The proliferation of gamma delta T cells were observed by plotting growth curve. The purity of gamma delta T cells were detected by flow cytometry and immunohistochemistry.


RESULTS
Stimulated by anti-TCR gamma delta mAb and IL-2, gamma delta T cells of peripheral blood and BALF could proliferate for a few days. The purity of gamma delta T cells obtained by Attack-panning method was 81%-99%.


CONCLUSIONS
The attack-panning method can get pure gamma delta T cells from peripheral blood and BALF.
Li
Chao-qian
CQ
Department of Respiratory Medicine, First Affiliated Hospital, Guangxi Medical University, Nanning 530021, China. lichaoqian@yahoo.com.cn
Xu
Yong-jian
YJ
Chen
Shi-xin
SX
Ni
Wang
W
Zhong
Xiao-ning
XN
Yang
Dan-lei
DL
Liu
Xian-sheng
XS
Zhang
Zhen-xiang
ZX
D000818:Animals; D000911:Antibodies, Monoclonal / Q000494:pharmacology; D001992:Bronchoalveolar Lavage Fluid / Q000166:cytology* / Q000276:immunology; D002455:Cell Division; D002478:Cells, Cultured; D003165:Complement System Proteins / Q000276:immunology; D003602:Cytotoxicity, Immunologic; D007376:Interleukin-2 / Q000494:pharmacology; D018655:Lymphocyte Count; D008297:Male; D051381:Rats; D017208:Rats, Wistar; D016693:Receptors, Antigen, T-Cell, alpha-beta / Q000276:immunology / Q000378:metabolism; D016692:Receptors, Antigen, T-Cell, gamma-delta / Q000276:immunology / Q000378:metabolism*; D016176:T-Lymphocyte Subsets / Q000166:cytology / Q000276:immunology*; D013602:T-Lymphocytes, Cytotoxic / Q000166:cytology / Q000276:immunology*
D000911:Antibodies, Monoclonal; D007376:Interleukin-2; D016693:Receptors, Antigen, T-Cell, alpha-beta; D016692:Receptors, Antigen, T-Cell, gamma-delta; D003165:Complement System Proteins