Improved IL-2 detection for determination of helper T lymphocyte precursor frequency in limiting dilution assay.
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.
Winandy
M
M
Université Catholique de Louvian, Laboratoire d'Immunohématologie, Brussels, Belgium.
Lewalle
P
P
Deneys
V
V
Ferrant
A
A
De Bruyère
M
M
D000818:Animals; D002478:Cells, Cultured; D006224:Cricetinae; D003470:Culture Media; D006086:Graft vs Host Disease / Q000097:blood; D006801:Humans; D007376:Interleukin-2 / Q000032:analysis* / Q000378:metabolism; D015847:Interleukin-4 / Q000494:pharmacology; D007668:Kidney / Q000378:metabolism / Q000502:physiology; D051379:Mice; D011237:Predictive Value of Tests; D015203:Reproducibility of Results; D012680:Sensitivity and Specificity; D013602:T-Lymphocytes, Cytotoxic / Q000166:cytology; D006377:T-Lymphocytes, Helper-Inducer / Q000166:cytology*; D014162:Transfection
D003470:Culture Media; D007376:Interleukin-2; D015847:Interleukin-4