| Graft-versus-host disease (GVHD) is the major complication after allogeneic bone marrow transplantation(allo-BMT) and is initiated by alloreactive donor T cells recongnizing foreign histocompatibility antigens of the host. Severe aGVHD is generally deadly. Nowadays, many measures are taken to prevent aGVHD, which include administration of G-CSF to donors. In our department, in last 7 days before bone marrow harvest, donors were given G-CSF by a dose of 250 u g per day. Compared with those patients whose donors were untreated with G-CSF, the patient undergoing BMT after their donors administrated G-CSF developed less severe aGVHD. It is clear that during the process of aGVHD, T lymphocytes act as a key role. To clarify the mechanism of G-CSF administration to donors reducing the incidence of aGVHD, in our study, assay of expression of IL-2 in lymphocytes was exerted. And procedure was designed as followed: from 2000 Oct to 2001 Apr, 10 leukemia patients were treated with Allo-BMT consecutively. Then-donors were given G-CSF by a dose of 250 u g per day, lasting for 7 days. At the 8th day, while donors being anesthetized, donors' bone marrow was harvested, and right after this, harvested bone marrow was implanted torecipients. 5ml of the donors' bone marrow and peripheral blood was gathered before and after G-CSF administration, respectively. These peripheral blood and bone marrow samples were separated and mononuclear cells were gathered , their concentration being adjusted to 1 X 106 /ml with 10% FCS-RPMI1640. PMA and lonomycin were administrated to promote these cells to produce BL-2. After this, FITC-Anti-CD3 and PE-Anti-IL-2 were used to combine with CDS and IL-2 expressed on surface of cells and in the plasma of the cells, respectively. Fixed by PBS, the samples were ready for FCM analysis. The results were obtained after FCM analysis, which showed that the expression of IL-2 in lymphocytes both from bone marrow and peripheral blood of 10 normal donors decreased significantly after G-CSF administration: The mean percentage of cells expressing IL-2 hi donors' bone marrow donors' peripheral blood was 15.67+6.19% before G-CSF administration and 3.69 ?3.25% after G-CSF administration. The mean percentage of cells expressing IL-2 hi donors' peripheral blood was 17.96+8.63% before G-CSF administration and 3.29 ?.0% after G-CSF admrnistration. The results suggest that maybe decrease of incidence of aGVHD in patients whose donors received G-CSF administration is correlated with such changes in donors' lymphocytes of both bone marrow and peripheral blood as decreased expression of DL-2 hi them, ie, a series of secretion function changes of lymphocytes reduce the incidence of aGVHD subsequently. Decrease of the expression of IL-2 in lymphocytes after administration of G-CSF support this theory: it is the subset of T lymphocytes converting from Thl to Th2, which bring about decrease of the expression of BL-2, that reduce the incidence of aGVHD finally.10Whereas it is reported that while aGVHD develops, NK cells also play an important role. NK cells meditate aGVHD through its toxicity and cytokine they produce after they are activated. To clarify whether the administration of G-CSF to donors will lead to changes of NK cells and if so, what changes will appear, and whether these changes finally affect the incidence of aGVHD, from 2001 Aug to 2002 Feb, 8 leukemia patients were treated with Allo-BMT consecutively in our department. Their donors were given G-CSF by a dose of 250 u g per day, lasting for 7 days. 5 ml of the donors' peripheral blood was gathered before and after G-CSF administration, respectively, for assay of the percentage of NK cells. Double-colored fluorescence-labeled monoclonal antibodies (PE-Anti-CD56^ FITC-Anti-CD3) were used to identify NK cells in the peripheral blood samples. After reaction, the samples were fixed by PBS, and later for FCM analysis. The results were obtained after FCM analysis, which showed that the percentage of NK cells in the peripheral blood decreased sign...
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