Improved IL-2 detection for determination of helper T lymphocyte precursor frequency in limiting dilution assay
(1998) Journal of Immunological Methods — Vol. 215, n° 1-2, p. 81-94 (1998)
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- 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. (C) 1998 Elsevier Science B.V. All rights reserved.
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Winandy, M., Ferrant, A., Lewalle, P., Deneys, V., & De Bruyère, M. (1998). Improved IL-2 detection for determination of helper T lymphocyte precursor frequency in limiting dilution assay. Journal of Immunological Methods, 215(1-2), 81-94. https://doi.org/10.1016/S0022-1759(98)00065-9 (Original work published 1998)