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© The Rockefeller University Press, 0022-1007/1996/12/2185/ $5.00
The Journal of Experimental Medicine, Volume 184, Number 6, December 1, 1996 2185-2196

Articles

Dendritic Cell Development in Culture from Thymic Precursor Cells in the Absence of Granulocyte/Macrophage Colony-stimulating Factor

Dolores Saunders*, Karen Lucas*, Jamila Ismaili*, Li Wu*, Eugene Maraskovsky{ddagger}, Ashley Dunn§, and Ken Shortman*

From * The Walter and Eliza Hall Institute of Medical Research, Melbourne, Victoria 3050, Australia; {ddagger} Immunex Research and Development Corporation, Seattle, Washington 98101; and § Ludwig Institute for Cancer Research, Melbourne, Victoria 3050, Australia

The earliest lymphoid precursor population in the adult mouse thymus had previously been shown to produce not only T cells, but also dendritic cell (DC) progeny on transfer to irradiated recipients. In this study, culture of these isolated thymic precursors with a mixture of cytokines induced them to proliferate and to differentiate to DC, but not to T lineage cells. At least 70% of the individual precursors had the capacity to form DC. The resultant DC were as effective as normal thymic DC in the functional test of T cell stimulation in mixed leukocyte cultures. The cultured DC also expressed high levels of class I and class II major histocompatibility complex, together with CD11c, DEC-205, CD80, and CD86, markers characteristic of mature DC in general. However, they did not express CD8{alpha} or BP-1, markers characteristic of normal thymic DC. The optimized mixture of five to seven cytokines required for DC development from these thymic precursors did not include granulocyte/macrophage colony stimulating factor (GM-CSF), usually required for DC development in culture. The addition of anti–GM-CSF antibody or the use of precursors from GM-CSF–deficient mice did not prevent DC development. Addition of GM-CSF was without effect on DC yield when interleukin (IL) 3 and IL-7 were present, although some stimulation by GM-CSF was noted in their absence. In contrast, DC development was enhanced by addition of the Flt3/Flk2 ligand, in line with the effects of the administration of this cytokine in vivo. The results indicate that the development of a particular lineage of DC, probably those of lymphoid precursor origin, may be independent of the myeloid hormone GM-CSF.

Address correspondence to Dr. K. Shortman, The Walter and Eliza Hall Institute of Medical Research, Post Office Royal Melbourne Hospital, Victoria 3050, Australia.

This work was supported by a Human Frontier Science Program grant and by the National Health and Medical Research Council, Australia.

1Abbreviations used in this paper: CD40L, CD40 ligand; DC, dendritic cells; Flt3L, Flt3/Flk2 ligand; SCF, stem cell factor.


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