Winandy M, Lewalle P, Deneys V, Ferrant A, De Bruyère M
Université Catholique de Louvian, Laboratoire d'Immunohématologie, Brussels, Belgium.
J Immunol Methods. 1998 Jun 1;215(1-2):81-94. doi: 10.1016/s0022-1759(98)00065-9.
In the context of allogeneic bone marrow transplantation, an accurate estimate of the risk of developing graft-versus-host disease (GVHD) is of major interest. The pre-transplant frequency of donor's helper T-lymphocyte precursors (HTLp) directed against host's antigens may be helpful in predicting this risk. This technique relies on an indirect measurement of interleukin-2 (IL-2) secreted by the HTLp, as assessed by the proliferation of an IL-2 dependent cell line. Many authors use the murine CTLL-2 cell line in this assay, but these cells do not respond to the presence of minute amounts of IL-2 in the culture medium, and thus do not discriminate between the absence or the presence of very low levels of IL-2. We therefore decided to compare CTLL-2 with another IL-2 dependent cell line, the murine A9.12 cell line. A comparison was made using serial dilutions of recombinant human IL-2, limiting dilutions of baby hamster kidney (BHK) cells transfected with human IL-2 gene and in the context of clinical tests performed for the detection of pre-transplant HTLp. Both the sensitivity and reliability of the tests were better using A9.12. We conclude that the A9.12 cell line might be a more suitable tool for pre-transplant HTLp determinations before allogeneic bone marrow transplantation or whenever low IL-2 levels are to be measured.
在异基因骨髓移植的背景下，准确估计发生移植物抗宿主病（GVHD）的风险备受关注。供体针对宿主抗原的辅助性T淋巴细胞前体（HTLp）的移植前频率可能有助于预测这种风险。该技术依赖于对HTLp分泌的白细胞介素-2（IL-2）的间接测量，通过IL-2依赖细胞系的增殖来评估。许多作者在该测定中使用小鼠CTLL-2细胞系，但这些细胞对培养基中微量IL-2的存在无反应，因此无法区分IL-2极低水平的有无。因此，我们决定将CTLL-2与另一种IL-2依赖细胞系——小鼠A9.12细胞系进行比较。使用重组人IL-2的系列稀释液、转染人IL-2基因的幼仓鼠肾（BHK）细胞的有限稀释液，并在检测移植前HTLp的临床试验背景下进行了比较。使用A9.12时，检测的灵敏度和可靠性都更好。我们得出结论，A9.12细胞系可能是在异基因骨髓移植前或任何需要测量低IL-2水平时，用于移植前HTLp测定的更合适工具。
