title: [Cytokine production and clonal analysis of memory γδ T cells from patients with tuberculous pleurisy induced by bacille calmette guerin].
abstract: OBJECTIVE
  To evaluate cytokine production and expression of γδ T cells within pleural fluid cells (PFCs) from patients with tuberculous pleurisy following bacille calmette guerin (BCG) stimulation.
  
  
  METHODS
  PFCs were isolated from patients with tuberculous pleurisy, and assessed for cytokine production, cell subpopulation, phenotype and characterization of T cell receptors after stimulation with BCG. The positive PCR products were further labeled with fluorescence and analyzed by genescan technique to determine the CDR3 size and evaluate the clonality of the detectable TCR Vγ and Vδ T cells.
  
  
  RESULTS
  Following stimulation with BCG, the positivity of interferon-γ (IFN-γ)-producing CD(4) T cells and γδ T cells were 0.38% and 5.35%, respectively. Phenotypic analysis indicated that the majority of IFN-γ(+)γδ(+) T cells expressed CD(45RO)(+) (73.5%). In addition, δ(2) T cells produced IFN-γ (11.1%) and TNF-α (25.5%). After expansion with BCG for 3 weeks, cells were harvested and mRNA extracted and RT-PCR conducted to amplify cDNA with 3 primers for Vγ and 8 primers for Vδ. The results indicated that BCG selectively expanded δ(2) T cells with oligoclonal peak in Vδ(2) cells.
  
  
  CONCLUSIONS
  BCG induced memory γδ and δ(2) T cells to produce cytokines in PFCs. Genescan analysis showed that Vδ(2) displayed oligoclonality.
authors.affiliation: Department of Immunology, Zhongshan School of Medicine, Sun Yat-sen University, Key Laboratory of Tropical Disease Control Research of Ministry of Education, Guangzhou 510080, China.
authors.forename: Li
authors.initials: L
authors.lastname: Li
authors.forename: Dan
authors.initials: D
authors.lastname: Qiao
authors.forename: Sui-Hua
authors.initials: SH
authors.lastname: Lao
authors.forename: Shao-Hua
authors.initials: SH
authors.lastname: Chen
authors.forename: Yang-Qiu
authors.initials: YQ
authors.lastname: Li
authors.forename: Chang-You
authors.initials: CY
authors.lastname: Wu
mesh_terms: D006801:Humans; D007156:Immunologic Memory; D007371:Interferon-gamma / Q000378:metabolism; D009163:Mycobacterium bovis / Q000276:immunology*; D016692:Receptors, Antigen, T-Cell, gamma-delta / Q000378:metabolism; D018121:Receptors, Cytokine / Q000378:metabolism; D016176:T-Lymphocyte Subsets / Q000276:immunology; D013601:T-Lymphocytes / Q000276:immunology*; D014396:Tuberculosis, Pleural / Q000276:immunology*; D014409:Tumor Necrosis Factor-alpha / Q000378:metabolism
chemical_list: D016692:Receptors, Antigen, T-Cell, gamma-delta; D018121:Receptors, Cytokine; D014409:Tumor Necrosis Factor-alpha; D007371:Interferon-gamma