title: [In vitro effects of mesenchymal stem cells on secreting function of T lymphocytes and CD4⁺CD25⁺ T cells from patients with immune thrombocytopenia].
abstract: OBJECTIVE
  To analyze in vitro the effect of mesenchymal stem cells (MSCs) on secreting cytokines by T lymphocytes and ratio of CD4⁺CD25⁺ T cells from patients with immune thrombocytopenia (ITP).
  
  
  METHODS
  Human bone marrow-derived MSCs were isolated by Ficoll Hypaque and cultured for proliferating to passage cells. Allogeneic T lymphocytes of health adults and ITP patients were isolated from peripheral blood by Ficoll Hypaque and nylon cotton column, and the ratio of CD4⁺CD25⁺ T cells was detected by flow cytometry. Then the different amounts of 1 × 10⁴, 5 × 10⁴, 2 × 10⁵ MSCs per well treated with mitomycin as stromal feeder layers were co-cultured with above-mentioned T lymphocytes, 5 days after cocultivation, the ratio of CD4⁺CD25⁺ T cells was detected by flow cytometry and the levels of IL-2, IFN-γ, IL-4, IL-10 were measured by enzyme- linked immune sorbent assay (ELISA).
  
  
  RESULTS
  After co-cultured with 2 × 10⁵ MSCs for 5 days, the ratio of CD4⁺CD25⁺ T cells and CD4⁺CD25⁺/CD4⁺ were significantly higher than of separate T lymphocytes in ITP patients [(4.56 ± 0.70)% vs (2.24 ± 0.81)%, (9.91 ± 1.18)% vs (4.08 ± 1.17)%, respectively] (P<0.05). To compare with separate T lymphocytes in ITP patients, the cytokine concentrations of IL-2 and IFN-γ from the culture supernatants significantly reduced from (280.47 ± 17.33) pg/ml to (97.21 ± 12.07) pg/ml and from (129.33 ± 16.34) pg/ml to (72.75 ± 7.81) pg/ml, respectively. In contrast, the cytokine concentrations of IL-4 and IL-10 increased from (16.34 ± 2.60) pg/ml to (37.98 ± 4.05) pg/ml and from (54.78 ± 5.62) pg/ml to (113.77 ± 5.68) pg/ml, respectively.
  
  
  CONCLUSIONS
  MSCs significantly inhibited the cytokine levels of IL-2 and IFN-γ secreted by Th1 cells and promoted the releases of IL-4 and IL-10 by Th2 cells in ITP , thereby regulating the balance between Th1 and Th2 reaction, as well as up-regulating the expression of CD4⁺CD25⁺ T cells in vitro,then induced the immunologic tolerance of ITP.
authors.affiliation: Department of Hematology, the Central Hospital of Shengli Oil Field, Dongying 257000, China.
authors.forename: Xia
authors.initials: X
authors.lastname: Zhao
authors.forename: Hui-fang
authors.initials: HF
authors.lastname: Ding
authors.forename: Cheng-shan
authors.initials: CS
authors.lastname: Guo
authors.forename: Xi-jing
authors.initials: XJ
authors.lastname: Lu
authors.forename: Min
authors.initials: M
authors.lastname: Xu
authors.forename: Jian
authors.initials: J
authors.lastname: Xing
authors.forename: Fang
authors.initials: F
authors.lastname: Han
authors.forename: Liang
authors.initials: L
authors.lastname: Wang
authors.forename: Guang
authors.initials: G
authors.lastname: Lu
authors.forename: Guo-qiang
authors.initials: GQ
authors.lastname: Liu
mesh_terms: D000293:Adolescent; D000328:Adult; D015496:CD4-Positive T-Lymphocytes / Q000378:metabolism*; D002478:Cells, Cultured; D005260:Female; D005434:Flow Cytometry; D006801:Humans; D007371:Interferon-gamma / Q000378:metabolism; D016753:Interleukin-10 / Q000378:metabolism; D007376:Interleukin-2 / Q000378:metabolism; D015847:Interleukin-4 / Q000378:metabolism; D008297:Male; D059630:Mesenchymal Stem Cells / Q000166:cytology*; D008875:Middle Aged; D013921:Thrombocytopenia / Q000378:metabolism*; D055815:Young Adult
chemical_list: C573415:IFNG protein, human; C508609:IL10 protein, human; C508594:IL2 protein, human; C508598:IL4 protein, human; D007376:Interleukin-2; D016753:Interleukin-10; D015847:Interleukin-4; D007371:Interferon-gamma