In this study we examined by Northern blot analysis the expression of Raf-1 protooncogene in normal human peripheral blood leukocytes. Unlike thymocytes, circulating lymphocytes did not express appreciable levels of Raf-1 mRNA. In contrast, polymorphonuclear cells (PMN) had high levels of Raf-1 transcripts. Also density gradient separated monocytes showed Raf-1 mRNA but at lower levels compared to PMN. Expression of Raf-1 was constitutive inasmuch as it was not induced by the purification procedure. The half-life of Raf-1 mRNA in PMN was >4 h. Functional activation of PMN and monocytes with various stimuli (phorbol esters, tumor necrosis factor, colony stimulating factors, LPS) did not affect Raf-1 expression. By contrast, density gradient purified monocytes allowed to adhere to plastic for 1 h expressed augmented levels of Raf-1. Monocytes cultivated in suspension or allowed to adhere to plastic showed an half-life of Raf-1 transcripts of, respectively, more than 4 h and less than 30 min. Circulating lymphocytes stimulated with mitogens (PHA, conA, anti-CD3 antibodies, and Staphylococcus aureus) also expressed high levels of transcripts of this protooncogene. PHA-induced transcripts in lymphocytes had an half-life > 4 h. The pattern of expression of Raf-1 in resting and activated leukocytes suggests that this protooncogene may play a role in expression of differentiated functions and activation of these cells.
ASJC Scopus subject areas
- Cell Biology