Functional expression of native and chimeric human and murine IL-2 receptor p70 chains in an IL-3-dependent murine cell line.
Full length cDNAs encoding the human and murine p70 genes were isolated using polymerase chain reaction (PCR) and conventional cDNA library screening techniques, respectively. To validate their functional potential, expression vectors containing human, murine and chimeric human/murine p70 cDNAs were transfected into the murine IL-3-dependent cell line FDC-P1. Transfected cells expressed a combination of high and low-affinity IL-2 binding sites while parental cells displayed only the low-affinity sites associated with expression of endogenous p55 IL-2 receptor chains. The role of the transfected p70 chains in formation of the high-affinity receptor sites was confirmed by the finding that the species-specific inhibitory antibody TU27 blocked high-affinity binding to human p70 and chimeric human/murine p70-expressing cells while having no effect on cells transfected with the murine p70 cDNA construct. As consequences of the expression of the transfected p70 chains, the cells responded to IL-2 with increased levels of endogenous p55 receptor subunit and both short and long-term proliferation. These studies substantiate the role of the p70 receptor chain in functional responses to IL-2 and provide a model system for future dissection of the structure/function relationships of the IL-2 receptor.
Freimark
B D
BD
Central Research and Development Department, Glenolden Laboratory, DuPont, PA 19036.
Robb
R J
RJ
D000818:Animals; D000911:Antibodies, Monoclonal; D002455:Cell Division / Q000502:physiology; D002460:Cell Line; D005434:Flow Cytometry; D005822:Genetic Vectors / Q000502:physiology; D006801:Humans; D016130:Immunophenotyping; D007377:Interleukin-3 / Q000502:physiology*; D051379:Mice; D015375:Receptors, Interleukin-2 / Q000096:biosynthesis* / Q000235:genetics / Q000276:immunology; D011994:Recombinant Proteins / Q000096:biosynthesis / Q000235:genetics; D014162:Transfection / Q000235:genetics
D000911:Antibodies, Monoclonal; D007377:Interleukin-3; D015375:Receptors, Interleukin-2; D011994:Recombinant Proteins