SYNOPSIS Current techniques for lymphocyte transformation are evaluated and criticised. A simple technique, designed to meet these criticisms, is described in detail, with particular reference to lymphocyte separation and scoring methods.Techniques for inducing morphological transformation of lymphocytes to large basophilic blast forms by non-specific mitogens, e.g. phytohaemagglutinin or 'pokeweed', or by specific antigens such as tetanus toxoid, have been described by

P-araproteins-Hobbs NIEMC-JURA et al. (1964) found that after multiple passages the cells comprising the plasmacytoma could become considerably less differentiated, producing less paraprotein and growing faster. A few patients with obviously extensive rapidly growing myeloma similarly produce little or even no paraprotein, especially those who produce only Bence Jones protein.In lymphosarcomata and in chronic lymphatic leukaemia, on the other hand, it seems to be the rule that when paraprotein production occurs it involves only a small percentage of the tumour cells. MacKay et al. (1957) thought that the presence of macroglobulinaemia in such cases was a secondary phenomenon, and arose from a side-line of mutated cells. The presence of more than one paraprotein has been demonstrated in three of our six patients with lymphomata-an incidence that is in marked contrast to our findings in myelomatosis, five out of a total of 320 patients showing more than one paraprotein. These findings also agree with those of Imhof et al. (1966), and support the hypothesis that paraprotein production in the .lymphomata reflects unstable, mutating cell populations.Paraproteins are also found in association with other malignant diseases, but their frequency is no greater than would be expected (Hdllen, 1966). Moreover, their levels rarely correlate with the removal or progress of the cancer, and our own experience suggests that this largely occurs by chance.

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The sensitivity of lymphocytes to chlorambucil has been assessed by a simple in-vitro test which has been applied to the cells of normal controls and of patients with chronic lymphocytic leukaemia. The degree of sensitivity varied amongst the normal controls and in-vitro resistance of the lymphocytes in the patients was sometimes found in the absence of in-vivo experience of the drug. Resistance in-vitro tended to be associated with very high total peripheral bloodlymphocytecountsbutnotwiththeageofthepatient. Wheretheinformation was available the in-vitro sensitivity test agreed with the results of biochemical estimations of drug resistance and with the clinical responses to the drug. It is suggested that this test may have applications in patient

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