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Characterization of cell phenotype by a novel cDNA library subtraction
system: expression of CD8 alpha in a mast cell-derived interleukin-4-
dependent cell line
T Hara, N Harada, H Mitsui, T Miura, T Ishizaka and A Miyajima
Department of Molecular Biology, DNAX Research Institute of Molecular and
Cellular Biology, Palo Alto, CA 94304.
We have established a unique variant cell line, MC/9.IL-4, which
continuously proliferates in the presence of interleukin-4 (IL-4), from a
murine interleukin-3 (IL-3)-dependent mast cell line, MC/9 (referred to as
MC/9.IL-3). Compared with MC/9.IL-3 cells, MC/9.IL-4 cells are smaller,
lack cytoplasmic granules and metachromasia, carry a very small amount of
histamine, and express fewer high-affinity IgE receptors (IgERs) and IL-3
receptors. To further characterize MC/9.IL- 4, we developed a novel method
to enrich cell type-specific cDNAs by cDNA library subtraction and applied
it for MC/9.IL-3 versus MC/9.IL-4. Sequence analysis of cDNA clones
isolated by this technique showed that MC/9.IL-4 cells specifically express
CD8 alpha and expression of mast cell-specific proteases and major
histocompatibility complex class II (MHCII) is considerably decreased. It
was also noted that responsiveness to the IL-3-agonistic antibody F9 and
expression of the transcription factor GATA-2 is diminished in MC/9.IL-4,
indicating that MC/9.IL-4 have lost major characteristics of the bone
marrow-derived cultured mast cells. Because other T-cell marker antigens,
CD8 beta, CD4, Thy-1, were not detected on MC/9.IL-4 cells, MC/9.IL-4 cells
may represent an unknown class of hematopoietic cells that express CD8
alpha. This cell line will be useful in studies of IL-4-mediated signal
transduction, as well as transcriptional regulation of mast cell
characteristic genes. This study also demonstrates the effective use of the
cDNA library subtraction strategy to characterize unknown types of
hematopoietic cells at the molecular level.
Volume 84,
Issue 1,
pp. 189-199,
07/01/1994
Copyright © 1994 by The American Society of Hematology
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