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Induction of recombination activating gene expression in a human lymphoid
progenitor cell line: requirement of two separate signals from stromal
cells and cytokines
H Tagoh, H Kishi, A Okumura, T Kitagawa, T Nagata, K Mori and A Muraguchi
Department of Immunology, Faculty of Medicine, Toyama Medical and
Pharmaceutical University, Japan.
The activation and expression of recombination activating genes (RAGs)
plays critical roles in V(D)J gene recombination machinery and lymphocyte
development. We showed that RAG gene expression was induced in freshly
isolated human bone marrow cells and a human lymphoid progenitor cell line,
FL8.2.4.4, by coculture on a monolayer of a murine bone marrow-derived
stromal cell line, PA6, in the presence of a mixture of recombinant
cytokines. The RAG transcripts were detected 12 hours after initiation of
culture, and the increased level was sustained at 24 hours. Among
recombinant cytokines, interleukin-3 (IL- 3), IL-6, and IL-7, but not IL-2,
IL-4, stem cell factor (SCF), and granulocyte-macrophage colony-stimulating
factor (GM-CSF) could induce RAG-1 activation in FL8.2.4.4 cells, and a
significant synergistic effect between IL-3, IL-6, and IL-7 was observed.
Using a double chamber culture technique, it was shown that a cognate
interaction between FL8.2.4.4 cells and PA6 stromal cells was a
prerequisite for RAG-1 activation. Furthermore, RAG-1 transcripts were
induced in FL8.2.4.4 cells when they were cocultured on
paraformaldehyde-fixed PA6 stromal cells in the presence of cytokines.
These results suggest that two separate signals are both required for
induction of RAG-1 activation in lymphoid progenitors; one from the cell
surface molecule(s) on stromal cells, and the other from the recombinant
cytokine(s). Finally, we showed that expression of RAG mRNA in FL8.2.4.4
cells was concomitant with induction of recombinase activity. This system
may provide useful means for further understanding the mechanisms
controlling RAG activation and lymphocyte development in the human system.
Volume 88,
Issue 12,
pp. 4463-4473,
12/15/1996
Copyright © 1996 by The American Society of Hematology
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