Nucleic acid changes during lymphocyte transformation in vitro
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Abstract
Circulating blood lymphocytes (a) do not usually divide in vitro (Trowell,1965), and (b), are of importance in immunity (Gowans + McGregor,1965). Nowell in 1960 discovered that when a bean extract, PHA, was added to these cells in culture they were suddenly activated to transform and divide. Similar cytological changes are seen within lymphoid organs in the course of immune responses (Fagraeus, 1948; Scothorne,1957; Gowans,1962; Oort + Turk,1965). I have studied the nucleic acid changes occurring during in vitro transformation in order to gain some understanding of the cellular control mechanisms involved and of the immune significance of the phenomenon. The results of this study are presented from chapter IV onwards after initial chapters on the literature of lymphocyte transformation in vitro (chapter II), and on the methods which have been employed (chapter III). In chapter IV the quantitative nucleic acid changes occuring during transformation are described. Such measurements were designed to produce results which could be simply interpreted. The main questions which are asked are:- (a) How long will cells grow in culture without medium replenishment? (b) How much PHA must be added in order to obtain a given transformation response? (c) Is this amount of PHA proportional to the number of cells present, or the amount of serum in the culture? (d) How long must cultures be exposed to PHA in order that a subsequent transformation response will occur? However quantitative nucleic acid assay was not sufficiently sensitive to answer the further questions which arose, In chapter V measurements of the incorporation of [5-