2‐Mercaptoethanol acts at the restricted stage of interleukin‐2 dependent lymphocyte proliferation

Abstract

An interleukin-2 (IL-2) dependent murine cell line (TN-9) which could be grown continuously with the crude culture supernatant of concanavaline A-stimulated rat or mouse spleen cells could not synthesize DNA in the culture medium supplemented with partially purified or recombinant IL-2. The cell growth was restored by adding another factor obtained from the same crude culture supernatant. This factor, physicochemically inseparable from serum albumin, was also obtained from the culture medium added with 2-mercaptoethanol (2-ME) and incubated at 37 degrees C for 24 hr without the cells. By the experiments using semi-synchronized cell population, it was demonstrated that 2-ME or 2-ME carrying protein acted at the restricted process(es) of cell proliferation which occurred between IL-2-acting stage and the initiation of DNA synthesis.

Synthesis of Sterols and 5‐Lipoxygenase Products are Required for the G1‐S Phase Transition of Interleukin‐2‐Dependent Lymphocyte Proliferation

“…We confirmed this by flow cytometric analysis. TN-9, another IL-2-dependent cell line whose profile of cell cycle is well characterized (21,34), was used for this experiment. This cell line can be easily cultured in a defined serum-free medium (HB101) (35) supplemented with recombinant IL-2 (rIL-2) and 2-mercaptoethanol (2-ME).…”