Abstract
Lymphocytes may be stimulated to undergo transformation into blast cells by a variety of nonspecific chemical agents, or by antiglobulin or antilymphocyte antisera. Antigens act as mitogenic agents on lymphocytes from sensitized animals (1-3).
We have now observed that the action of various mitogens on human and rabbit peripheral lymphoeytes is markedly enhsnced in the presence of the reducing agents, l-cysteine, glutathione, or sulfite.
Rabbit peripheral lymphocytes were isolated, purified on a cotton-wool column and cultured as described previously (4). Human leukocytes were obtained from the blood of normal individuals by allowing the cells to settle in 10% Plasmagel (Laboratoire Roger Bellon, Neuilly, France). The leukocyte-rich layer was removed and the cells were washed several times with 20% decomplemented fetal calf serum in Eagle's minimal essential medium (MEM). Leukocytes were cultured (4) at a concentration of 106 cells/ml. Rabbit lymphoeytes were cultured for 48 hr and human leukocytes for 108 hr; 24 hr before harvest, 5 µCi of 3H-thymidine (6.7 Ci/mM) was added to the rabbit lymphocyte cultures and either 5 µCi or 2 µCi of 3H-thymidine to the human leukocyte cultures. Radioactivity incorporated into DNA was determined as described previously (4).
Footnotes
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↵2 Postdoctoral Fellow of the National Institutes of Health.
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↵3 Supported by University of California Training Grant HE-05677 (USPHS) and the Royal Australasian College of Physicians.
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↵1 This work was supported by grants from the National Science Foundation (GB-5424) and the National Institutes of Health.
- Copyright © 1970 by The American Association of Immunologists, Inc.
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