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Metabolic Responses of Separated Leucocytes to Phytohaemagglutinin: Effects of Anaerobiasis, Actinomycin D and Puromycin
Author(s) -
Rabinowitz Yale,
Schimo Inge,
Wilhite Betty A.
Publication year - 1968
Publication title -
british journal of haematology
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.907
H-Index - 186
eISSN - 1365-2141
pISSN - 0007-1048
DOI - 10.1111/j.1365-2141.1968.tb01565.x
Subject(s) - puromycin , glycolysis , protein biosynthesis , phytohaemagglutinin , rna , biology , biochemistry , metabolism , dactinomycin , microbiology and biotechnology , cycloheximide , gene , in vitro
Metabolic changes needed for response to PHA were sought. Normal lymphocytes (N‐L), which respond strongly to PHA showed in 1.5–2.5 hour cultures sharp increases in RNA and protein synthesis, in glycolysis and O 2 consumption. Leukaemic lymphocytes (CLL‐L), which respond poorly to PHA, had higher initial rates of synthesis of RNA and protein, but lesser glycolytic activity than N‐L. Moderate increase in glycolysis occurred with PHA. Polymorphonuclear leucocytes (PMN), which do not transform with PHA, showed increased RNA synthesis and glycolysis in brief cultures, but protein synthesis decreased. N‐L and PMN showed a moderate increase in glycolysis in N 2 , while CLL‐L responded very strongly. Actinomycin D blocked RNA synthesis and puromycin blocked protein synthesis, but neither prevented initial stimulation of glycolysis by PHA. Glycolysis decreased, however, and blast‐cell formation was prevented in longer cultures with either antibiotic. Protein and RNA synthesis apparently are required to maintain the increased levels of glycolysis needed for cell transformation. It is suggested that PHA acts at allosteric sites on protein repressors or inducers of operator genes to activate a series of reactions which simulate those in immunological responses.