rhG-CSF动员的人外周血与骨髓不同比例混合对移植物免疫特性的影响

Effect of rhG-CSF mobilized peripheral blood and bone marrow after being mixed in different proportions on immunologic properties of grafts

目的探讨rhG-CSF动员的外周血采集物(G-PB)和骨髓(G-BM)不同比例混合对移植物免疫学特性的影响。方法用流式细胞仪测定G-PB和G-BM中T细胞亚群、树突状细胞(DC)亚群的数量,据有核细胞数将G-PB/G-BM按2∶11、∶11、∶2混合,计算混合移植物的免疫细胞组成;用MTT法和夹心ELISA法测定T细胞增殖能力和IL-4、IFN-γ的分泌。结果3种G-PB/G-BM混合移植物的T细胞增殖能力均低于未动员骨髓(NG-BM)和G-PB(P<0.05)。3种混合移植物与NG-BM、G-BM和G-PB在淋巴细胞、CD3+、CD4+、CD8+T细胞组成上存在差别(P<0.05);3种混合移植物的CD4/CD8比例都高于NG-BM和G-BM(P<0.01),DC1/DC2比值均低于NG-BM(P<0.01),IL-4/IFN-γ的比值均低于G-BM(P<0.05)。在输入相同CD34+细胞以保证植入的前提下,应用3种混合移植物进行移植时输入的Ⅰ型因子及Ⅰ型(Th1/Tc1)和Ⅱ型(Th2/Tc2)T细胞的数量介于NG-BM、G-BM和G-PB之间。结论G-PB/G-BM混合后免疫特性的改变可能在HLA不合移植中发挥作用。

Objective To explore the immunologic composition and T cell function of rhG-CSF mobilized peripheral blood grafts （G-PB） and rhG-CSF primed bone marrow grafts （G-BM） after being mixed in different proportions. Methods T cell subgroups, dendritic cells （DC） and its subsets in G-PB and G-BM were analyzed by multicolor flow cytometry. Immunologic composition of G-PB and G-BM after mixture in different proportions were calculated, lymphocyte proliferation ability and quantities of interleukin-4（ IL-4）, interferon-γ（IFN-γ） secretion by T cells in mixture grafts were determined using MTr and sandwich ELISA assays. Results After co-incubation of G-PB with G-BM in different proportions, T cell proliferation in mixture grafts was lower than that of NG-BM and G-PB（ P 〈 0.05）. There was significant difference of lymphocytes, CD3^＋ , CD4^＋ and CD8^＋ T cells between NG-BM, G-BM, G-PB and the mixture grafts （P 〈0.05）. The ratios of CD4/CD8 in mixture grafts were significantly higher than NG-BM plus G-BM （ P 〈 0.01 ）, whereas the ratios of DC1/DC2 were significantly lower than NG-BM （P 〈 0.01 ）, the ratios of IL-4/IFN-γ were siginificantly lower than GBM （P 〈 0.05）. The total quantities of type Ⅰ, type Ⅱcytokines and the counts of T1 （Th1/Tc1）, T2 （Th2/Tc2） cells infused in the mixture grafts were comparable when 2 × 10^6 CD34^＋ cells/kg were transplanted to ensure engraftment; both of which （except for type Ⅱcytokines） were either lower or higher than NG-BM, G-BM and G-PB; the type Ⅱcytokines were lower than NG-BM, G-BM and G-PB, while there was no statistical significace. Conclusion The alteration of immunologic composition and T cell function of G-PB and G-BM grafts after co-incubation in different proportions may contribute to the favorable outcome of patients transplanted with HLA mismatched haplotype donors.