Discrete stages of human thymocyte activation and maturation in vitro: correlation between phenotype and function

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Abstract
Using light scatter, flow cytometric and functional analyses, three major stages in the in vitro activation of human thymocytes were defined. The earliest stage (day 0–2) was characterized by the induction of the interleukin 2 (IL2) and transferrin receptors, the T cell lineage specific Tal antigen, and increased reactivity with anti‐T8 antibody. At this time, major changes in nuclear morphology but not cell size were observed. Early‐stage thymocytes were immature with regard to T3 and cytotoxic capacities. The second stage (day 3–4) of in vitro culture was distinguished by loss of T6, maximal activation (both in cell size and nuclear morphology) and maximal expression of both transferrin and IL2 receptors. At this stage, nearly all thymocytes expressed T3, 30–70% of thymocytes were T4+T8+, and functionally, only small increases in cytotoxic capacities were observed. The third stage of maturation (day 5–7) represented thymocytes with reduced levels of activation as measured by forward and right angle light scatter analysis and declining IL 2 and transferrin receptor expression. However, these thymocytes exhibited high levels of T3 antigen density, loss of T4, T8 coexpression and pronounced cytotoxic and detectable inducer function capabilities.