Colchicine ultrasensitivity of peripheral-blood lymphocytes from patients with non-Hodgkin's lymphoma.

Incubation for 20 h in low concentrations of colchicine has been shown to kill the peripheral blood lymphocytes (PBL) of patients with chronic lymphocytic leukaemia (CLL), whereas at least a 10,000 x higher concentration of colchicine is required to kill lymphocytes from a normal donor. This ultrasensitivity of CLL lymphocytes to low doses of colchicine was confirmed in 19/20 PBL samples, 5/6 lymph nodes, and in the one totally replaced marrow studied. PBL from 75 patients with non-Hodgkin's lymphoma (NHDL) were examined for colchicine ultrasensitive (CUS) cells similar to those found in CLL. All the patients had less than 5 x 10(9)/1 morphologically normal circulating lymphocytes. PBL from 45 healthy donors and 39 patients with diseases other than leukaemia or lymphoma were used as controls. CUS cells were detected in 24 (32%) of the 75 patients. The CUS cells were considered to represent blood involvement with malignant lymphocytes for three reasons. First, there was an association with marrow involvement (P less than 0.05) which usually accompanies involvement of the blood with morphologically abnormal cells. Secondly, 23 (77%) of the 30 involved lymph nodes, marrows and spleens studied were CUS. Thirdly, there was a close correlation with the presence of a monoclone of B lymphocytes demonstrated by surface markers (P less than 0.01).

Abnormal peripheral blood lymphocytes and bone marrow infiltration in non-Hodgkin's lymphoma.

The blood lymphocytes of 21 normal blood specimens, 20 patients with various malignant disorders and 63 patients with non-Hodgkin's disease lymphoma, all with lymphocyte counts below 6 x 10(9)/l, were examined by sheep red cell rosetting, fluorescent antisera to surface immunoglobulin and for sensitivity to colchicine. The results were correlated with bone marrow infiltration and lymph node histological findings. The ratio of kappa to lambda light chains in the surface immunglobulin was used to determine if an abnormal clone of lymphocytes was present. In the normals and the non lymphoma controls the expected normal ratio of approximately 2K to 1 lambda was found with a narrow spread. In non-Hodgkin's lymphoma cases there was a wide spread of ratios with half the cases outside the range of the controls. These cases were considered to have a clone of abnormal lymphocytes in the blood although their routine blood and differential counts were usually normal. There was a very significant correlation of the presence of a clone with ultrasensitivity to colchicine and with involvement of the bone marrow. 81% of the cases with a histological diagnosis of well-differentiated diffuse lymphocytic lymphoma, 64% of those with poorly differentiated diffuse but only 22% of those with diffuse histiocytic had an abnormal lymphocyte clone demonstrated by an abnormal K:lambda ratio.

The effect of dibromodulcitol on resting and dividing lymphoid cells.

The effect of dibromodulcitol (DBD) on the incorporation of labelled precursors into DNA and RNA fractions of PHA-stimulated human lymphocytes and of P388F lymphoma cells at various stages of their growth was studied. Both cell systems showed sensitivity to the drug within the concentration rage of 1-10 mug/ml. When DBD was added before phytohaemagglutinin (PHA), h.han RNA. In contrast, by adding DBD after PHA, RNA labelling was much more inhibited than DNA. In the latter case, the decrease in DNA labelling occurred only 24 h after drug treatment whereas RNA labelling was decreased 1 h after treatment. Levels of DBD which normally produced 30% inhibition in plating efficiency of P388F lymphoma cells affected uridine-5-T incorporation to a different extent at different stages of growth of the culture. Enhanced RNA labelling occurred in early exponential stage while at later stages of growth, RNA synthesis was depressed.

Separation of abnormal cells in the peripheral blood by means of the zonal centrifuge.

The basic principles of rate centrifugation in the large volume zonal centrifuge and its application to the separation of abnormal or rarely occurring cells in the blood are described. Details of the method, gradient media, and collection and examination of the fractions obtained are given. A description of the cells found in normal human blood, in a subject undergoing known immunological stimulus, and in glandular fever and Hodgkin's disease is given and the implications are discussed.

Quantitation of lymphocyte transformation using radioactive iododeoxyuridine.

A method is described of measuring the response of human lymphocytes to phytohaemagglutinin stimulation using iododeoxyuridine labelled with the gamma-emitting isotope (125)I. The results indicate useful clinical applications.