title: [The isolation and expansion of gamma delta T cells in peripheral blood and bronchoalveolar lavage fluid].
abstract: 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.
authors.affiliation: Department of Respiratory Medicine, First Affiliated Hospital, Guangxi Medical University, Nanning 530021, China. lichaoqian@yahoo.com.cn
authors.forename: Chao-qian
authors.initials: CQ
authors.lastname: Li
authors.forename: Yong-jian
authors.initials: YJ
authors.lastname: Xu
authors.forename: Shi-xin
authors.initials: SX
authors.lastname: Chen
authors.forename: Wang
authors.initials: W
authors.lastname: Ni
authors.forename: Xiao-ning
authors.initials: XN
authors.lastname: Zhong
authors.forename: Dan-lei
authors.initials: DL
authors.lastname: Yang
authors.forename: Xian-sheng
authors.initials: XS
authors.lastname: Liu
authors.forename: Zhen-xiang
authors.initials: ZX
authors.lastname: Zhang
mesh_terms: 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*
chemical_list: D000911:Antibodies, Monoclonal; D007376:Interleukin-2; D016693:Receptors, Antigen, T-Cell, alpha-beta; D016692:Receptors, Antigen, T-Cell, gamma-delta; D003165:Complement System Proteins