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In vitro activation of bone marrow natural killer T cells of aplastic anemia patients / 中华血液学杂志
Chinese Journal of Hematology ; (12): 536-539, 2010.
Article in Chinese | WPRIM | ID: wpr-353595
ABSTRACT
<p><b>OBJECTIVE</b>To investigate the quantitative and qualitative changes of TCRVα24(+)Vβ11(+) natural killer T (NKT) cells from bone marrow (BM) of aplastic anemia (AA) after in vitro stimulation of α-galactosylceramide (α-Galcer).</p><p><b>METHODS</b>NKT cells in the bone marrow mononuclear cells (BMMNCs) from either AA patients or healthy controls were enumerated with flow cytometry. BMMNCs were cultured in RPMI1640 medium supplemented with either α-Galcer and rhIL-2 or α-Galcer, rhIL-2 and rhG-CSF. The proliferative capacity of NKT cells was determined by NKT cell numbers before and after in vitro culture. Expression of intracellular IFNγ and IL-4 in activated NKT cells was analyzed with flow cytometry.</p><p><b>RESULTS</b>In AA group, the percentage of NKT cells in BMMNCs was (0.19 ± 0.09)%. Addition of rhG-CSF into the α-Galcer/rhIL-2 culture medium resulted in significantly reduced expansion of NKT cells (67.45 ± 29.42-fold vs 79.91 ± 40.56 fold, P < 0.05). Meanwhile, addition of rhG-CSF reduced IFNγ positive NKT cells \[(37.45 ± 7.89)% vs (62.31 ± 14.67)%, P < 0.01\] and increased IL-4 positive NKT cells \[(55.11 ± 12.13)% vs (27.03 ± 9.88)%, P < 0.01\]. In healthy control group, the percentage of NKT cells in BMMNCs was (0.25 ± 0.12)%. Addition of rhG-CSF into the α-Galcer/rhIL-2 culture medium also significantly reduced expansion of NKT cells (97.91 ± 53.22-fold vs 119.58 ± 60.49-fold, P < 0.05), reduced IFNγ positive NKT cells \[(28.65 ± 10.63)% vs (50.87 ± 12.66)%, P < 0.01\], and increased IL-4 positive NKT cells \[(66.53 ± 14.96)% vs (31.11 ± 10.07)%, P < 0.01\].</p><p><b>CONCLUSION</b>Compared to those from healthy controls, BMMNCs from AA patiants have a reduced fraction of NKT cells, which possesses a decreased potential to expand in vitro in response to α-Galcer stimulation, and produce more IFNγ(+) NKT1 cells. rhG-CSF, in combination with α-Galcer, confers polarization of NKT cells towards IL-4(+) NKT2 subpopulation.</p>
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Full text: Available Index: WPRIM (Western Pacific) Main subject: Bone Marrow / Killer Cells, Natural / Interleukin-4 / Cell Biology / Natural Killer T-Cells / Anemia, Aplastic / Metabolism Type of study: Qualitative research Limits: Humans Language: Chinese Journal: Chinese Journal of Hematology Year: 2010 Type: Article

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Full text: Available Index: WPRIM (Western Pacific) Main subject: Bone Marrow / Killer Cells, Natural / Interleukin-4 / Cell Biology / Natural Killer T-Cells / Anemia, Aplastic / Metabolism Type of study: Qualitative research Limits: Humans Language: Chinese Journal: Chinese Journal of Hematology Year: 2010 Type: Article