title: [Development of a CFSE-based flow cytometry for evaluating EIAV-stimulated proliferation of T lymphocytes].
abstract: OBJECTIVE
  To develop a flow cytometry using (5-carboxyfluorescein diacetate succinmidyl ester, CFSE) to detect the proliferation of specific T lymphocytes from equine infectious anemia virus (EIAV).
  
  
  METHODS
  Peripheral blood mononuclear cells (PBMC) were isolated, stained with CFSE and incubated with EIAV for 5 days. After interacted with either CD4(+) or CD8(+) antibody, the cells were detected for proliferated population, which contained serially 2-fold reduced CFSE in CD4(+) and CD8(+) T lymphocytes.
  
  
  RESULTS
  The concentration of CFSE, and the type, concentration and reaction time of EIAV-specific antigens were tested and optimized to improve the sensitivity and specificity of this flow cytometry-based assay. PBMCs isolated from the horses infected with either virulent or vaccine strains of EIAV and health controls were analyzed to evaluate this technique. The proliferation rates of CD4(+) and CD8(+) T lymphocytes were observed in the cells from differently treated horses.
  
  
  CONCLUSIONS
  The CFSE-based flow cytometry is a reliable tool to analyze antigen-specific proliferation of lymphocytes, which plays an important role in evaluating the immune responses induced by EIAV and other viruses.
authors.affiliation: Division of Livestock Infectious Diseases, Harbin Veterinary Research Institute of Chinese Academy of Agricultural Sciences, Harbin 150001, China.
authors.forename: Yue-zhi
authors.initials: YZ
authors.lastname: Lin
authors.forename: Xi-lin
authors.initials: XL
authors.lastname: Deng
authors.forename: Nan
authors.initials: N
authors.lastname: Shen
authors.forename: Xiao-ling
authors.initials: XL
authors.lastname: Lü
authors.forename: Li-ping
authors.initials: LP
authors.lastname: Zhao
authors.forename: Xian-gang
authors.initials: XG
authors.lastname: Kong
authors.forename: Yi-ming
authors.initials: YM
authors.lastname: Shao
authors.forename: Jian-hua
authors.initials: JH
authors.lastname: Zhou
mesh_terms: D000818:Animals; D015496:CD4-Positive T-Lymphocytes / Q000166:cytology / Q000276:immunology / Q000821:virology; D018414:CD8-Positive T-Lymphocytes / Q000166:cytology / Q000276:immunology / Q000821:virology; D049109:Cell Proliferation; D004859:Equine Infectious Anemia / Q000276:immunology / Q000821:virology; D005434:Flow Cytometry / Q000379:methods*; D005452:Fluoresceins / Q000737:chemistry*; D006736:Horses; D004860:Infectious Anemia Virus, Equine / Q000502:physiology*; D007963:Leukocytes, Mononuclear / Q000166:cytology / Q000276:immunology / Q000821:virology; D013388:Succinimides / Q000737:chemistry*; D013601:T-Lymphocytes / Q000276:immunology* / Q000821:virology
chemical_list: C087165:5-(6)-carboxyfluorescein diacetate succinimidyl ester; D005452:Fluoresceins; D013388:Succinimides