title: [Identification of cell division of CD8+ NKT cells in vitro].
abstract: OBJECTIVE
  To measure the in vitro cell division ability of CD8(+);NKT cells by CFSE staining and flow cytometry(FCM).
  
  
  METHODS
  Fresh spleen lymphocytes of C57BL/J mice were separated and stained with CFSE, and then stimulated by ConA and LPS for 3 d, and by SEB for 5 d and 10 d respectively. The stimulated cells were harvested and analyzed for CD69 expression on the cell surface and the ability of cell division using FCM. The SEB-activated effector cells for 10 d further stimulated with IL-2 for the consecutive 10 days, and were analyzed for their cell division ability, CD69 expression and NKT cell subsets by FCM.
  
  
  RESULTS
  ConA, LPS and SEB stimulated the proliferation of spleen cells. ConA and LPS made the cells divide 3 times within 3 d, and increased CD69 expression up to 74.19% and 41.56% respectively. SEB made the cells divide 5 times within 5 d and 7 times within 10 d respectively, with increased CD69 expression of 32.09% and 48.66% respectively. Ten-day IL-2 stimulation of SEB-activated cells caused population expansion for 7 times with the CD8(+);NKT cell subsets significantly increased from 0.36% to 38.58% and CD69 expression significantly increased from 0.11% to 83.74%.
  
  
  CONCLUSIONS
  The SEB-activated CD8(+);NKT cells proliferated in vitro and their cell division capability could be determined by CFSE staining and FCM.
authors.affiliation: Department of Immunology, General Hospital of PLA, Beijing 100853, China. guoyelei@sohu.com
authors.forename: Ye-Lei
authors.initials: YL
authors.lastname: Guo
authors.forename: Yu
authors.initials: Y
authors.lastname: Chen
authors.forename: Jiang
authors.initials: J
authors.lastname: Zhong
authors.forename: Shi-Lun
authors.initials: SL
authors.lastname: Zhang
mesh_terms: D000818:Animals; D015703:Antigens, CD / Q000276:immunology / Q000378:metabolism; D000945:Antigens, Differentiation, T-Lymphocyte / Q000276:immunology / Q000378:metabolism; D018414:CD8-Positive T-Lymphocytes / Q000166:cytology* / Q000378:metabolism*; D002455:Cell Division / Q000187:drug effects / Q000502:physiology*; D002478:Cells, Cultured; D004768:Enterotoxins / Q000276:immunology / Q000633:toxicity; D005260:Female; D037181:Lectins, C-Type / Q000276:immunology / Q000378:metabolism; D008213:Lymphocyte Activation / Q000187:drug effects / Q000276:immunology; D051379:Mice; D008810:Mice, Inbred C57BL; D055611:Natural Killer T-Cells / Q000166:cytology* / Q000378:metabolism*; D013997:Time Factors; D000099192:CD69 Antigens
chemical_list: D015703:Antigens, CD; D000945:Antigens, Differentiation, T-Lymphocyte; D004768:Enterotoxins; D037181:Lectins, C-Type; D000099192:CD69 Antigens; C031818:enterotoxin B, staphylococcal