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.

摘要: We have established a unique variant cell line, MC/9.IL-4, which continuously proliferates in the presence of interleukin-4 (IL-4), from murine interleukin-3 (IL-3)-dependent mast 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 very small amount histamine, express fewer high-affinity IgE receptors (IgERs) IL-3 receptors. To further characterize MC/9.IL- 4, we developed novel method enrich type-specific cDNAs by cDNA library subtraction applied it for versus MC/9.IL-4. Sequence analysis clones isolated this technique showed that specifically CD8 alpha expression cell-specific proteases major histocompatibility complex class II (MHCII) is considerably decreased. It was also noted responsiveness IL-3-agonistic antibody F9 transcription factor GATA-2 diminished indicating lost characteristics bone marrow-derived cultured cells. Because other T-cell marker antigens, beta, CD4, Thy-1, were not detected on may represent an unknown hematopoietic alpha. This line will be useful studies IL-4-mediated signal transduction, well transcriptional regulation characteristic genes. study demonstrates effective use strategy types at molecular level.