Scanning electron microscopy of human lymphocytes during transformation and subsequent treatment with methotrexate

Preparations of human peripheral blood lymphocytes containing 72-87% T-cells and 10-16% B-cells were shown by scanning electron microscopy to consist almost exclusively of cells bearing numerous microvilli, whereas thymocytes were of mixed surface morphology, with both smooth and encrusted forms numerous. T-lymphocytes purified on long nylon columns were all covered with numerous short villi. Stimulation with phytohaemagglutinin for 2 days produced T-lymphoblasts almost exclusively, and as the T-cells enlarged the microvilli lengthened, the increase in length reaching 5-fold by day 3. Addition of sufficient methotrexate on day 3 to arrest proliferation (50 nM) caused progressive loss of microvilli from the cell surface, with the eventual production of large numbers of smooth cells, the surfaces of which later became pitted, followed by the complete dissolution of the cell. T-lymphocytes were shown to form rosettes with sheep erythrocytes through direct contact of the cell membranes over a significant area, but when, as a result of methotrexate treatment, the lymphocytes had become denuded of microvilli or had reached an advanced state of dissolution, rosettes were no longer formed.