title: [Effect of koumine on proliferation of murine CD4+ T cells purified by magnetic-activated cell sorting in vitro].
abstract: OBJECTIVE
  To assess the separation efficiency of magnetic-activated cell sorting in the purification of CD4+ T cells from murine spleen, and observe the effects of koumine on the proliferation of the separated cells.
  
  
  METHODS
  CD4+ T cells were isolated from murine spleen by magnetic-activated cell sorting (MiniMACS). Fluorescence-activated cell sortering was employed to determine the purity of CD4+ T cells before and after the separation procedure followed by evaluation of the cell viability using trypan blue staining. Concanavalin A- (ConA, 5 microg/ml) or phytahematoagglutinin (PHA,1 mg/ml)-induced murine T cells were treated with different concentrations of koumine (10-320 microg/ml), and their proliferation was determined by MTT colorimetry, and enzyme-linked immunosorbent assay was used to measure IL-2 level in the cell culture supernatant.
  
  
  RESULTS
  The purity of CD4+ T cells reached (90.3+/-5.8)% after the purification with a cell viability of (94.9+/-3.6)%. Koumine (20-320 microg/ml) dose-dependently inhibited ConA- or PHA-induced proliferation of murine lymphocytes as compared with the controls (P<0.05). Koumine (20, 100, and 200 microg/ml) significantly decreased the level of IL-2 in comparison with the control group (P<0.05).
  
  
  CONCLUSIONS
  CD4+ T cells of high purity can be obtained from murine spleen using MiniMACS without impairing the viability of the cells. Koumine significantly inhibits the proliferation of murine CD4+ T cells due to its immunosuppressive effect and inhibition of IL-2 secretion.
authors.affiliation: Department of Pharmacy, Zhujiang Hospital, Southern Medical University, Guangzhou 510282, China. wangzr@fimmu.com
authors.forename: Zhi-rui
authors.initials: ZR
authors.lastname: Wang
authors.forename: Chang-quan
authors.initials: CQ
authors.lastname: Huang
authors.forename: Zhong-yi
authors.initials: ZY
authors.lastname: Zhang
authors.forename: Lan-lan
authors.initials: LL
authors.lastname: Zhang
authors.forename: Jing-ming
authors.initials: JM
authors.lastname: Lin
mesh_terms: D000818:Animals; D015496:CD4-Positive T-Lymphocytes / Q000166:cytology*; D049109:Cell Proliferation / Q000187:drug effects*; D002469:Cell Separation / Q000379:methods*; D002478:Cells, Cultured; D005260:Female; D031450:Gelsemium / Q000737:chemistry*; D026121:Indole Alkaloids / Q000494:pharmacology*; D008280:Magnetics; D008297:Male; D051379:Mice; D008807:Mice, Inbred BALB C
chemical_list: D026121:Indole Alkaloids; C515085:koumine