Stereological data of phytohaemagglutinin (PHA)-activated human T-lymphocytes were recorded at intervals (12 to 72 h) together with biochemical (isotope-uptake, lymphotoxin-release) and morphological measurements. About 98% of the cells were activated 12 h after PHA-stimulation. The activation phase lasted less than 48 h, i.e., cells entering the activation phase within 12 h were at their activation maximum by 48 h. The activated cell increased in size. The nuclear/cytoplasmic-ratio decreased. Most of the cytoplasmic organelles developed in phase with the increase of cytoplasmic volume. After 48 h, mitotic figures were frequently seen. Due to the increasing number of secondary, activated daughter cells, parameters of most cytoplasmic components declined between 48 and 72 h. Structural changes in the nucleus preceded the 3H-leucine uptake, which had not reached its maximum after 72 h of incubation. The 3H-leucine uptake started as early as 12 h after culture initiation, and its increase was proportional to the increasing polyribosome density. No maximum uptake was reached up to 72 h, but the development of structural components related to this uptake was at its maximum at the end of the activation phase (48 h). The formation of bound ribosomes occurred subsequent to the enlargement of the surface of the rough endoplasmic reticulum. Initial polysome formation occurred at the expense of existing free ribosomes.

T-lymphocytes derived from human peripheral blood and passed through a nylon-wool column, were employed to develop and test a new stereological model system for free spherical cells, allowing a quantitative characterization of the cell and its components at the ultrastructural level. Electron micrographs were recorded in a hierarchical manner at three different levels of magnification and subjected to point counting procedures. The resulting parameters were expressed in relation to various reference compartments, both absolute and relative. Results indicated that the average volume of a small, non-activated T-lymphocyte was 103.8 micron3, the nuclear volume 47.5 micron3 and the cytoplasmic volume 55.9 micron3. On the average, the cytoplasm contained 30 mitochondria, 0.7 micron3 RER-cisternae, 0.2 micron3 cisternae and vesicles of the Golgi apparatus and about 231,000 free ribosomes (most of them single). The ratio of eu- to heterochromatin volume was 0.5. The design and application of the stereological model system are discussed with regard to dynamic studies of a variety of free cells, such as macrophages, neutrophilic granulocytes and various lymphocytes.

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