By C. Garrison Fathman
Isolation Characterization, and usage of T Lymphocyte Clones.
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A. (1978b). Τ cell growth factor: Parameters of production and a quantitative microassay for activity. J. Immunol. 120, 2027. , Union, Ν. , Baker, P. , and Smith, K. A. (1979). The in vitro generation and sustained culture of nude mouse cytolytic T-lymphocytes. J. Exp. Med. 149, 1460. , Smith, Κ. , and Watson, J. D. (1980a). Biochemical and biological characterization of lymphocyte regulatory molecules. II. Purification of a class of rat and human lymphokines. J. Immunol. 124, 1954. , and Watson, J.
1978b). In addition to being instrumental for assessment of IL-2 activity following various biochemical separations and modifications, the IL-2 microassay remains as the only unequivocal assay for the presence of IL-2 activity. The IL-2 assay is based solely on the factor requirement of cultured Τ cell lines. Τ cells harvested from IL-2-dependent culture, washed and placed back in culture in the absence of IL-2, invariably die within 24 hr. By using tritiated 3 thymidine ( H-TdR) incorporation as an index on cultured Τ cell replication, the IL-2 microassay provides a highly reproducible and quantitative indication of the amount of IL-2 activity present in a given sample of crude or biochemically manipulated CM.
These results, in addition to documenting successful isolation and translation of producer cell, human IL-2 message, also lend evidence that posttranslational processing may not be necessary for synthesis of biologically active IL-2, since glycosylation is not possible in reticulocyte systems. 5% by volume) did not stimulate CTLL cell replication, whereas lesser concentrations did. 001 οοϊ αϊ μΙ/ΙΟ 3 CTLL [ο ioö CELLS Fig. 8. IL-2 microassay data gathered from titration of rabbit reticulosate translation product ( A).