Observations on prostaglandins in normal and leukemic human lymphocytes

Prostaglandins E (PGE) and F2 alpha (PGF2 alpha) were measured in lymphocytes of normal subjects, children with acute lymphocytic leukemia (ALL), and adults with chronic lymphocytic leukemia (CLL). In ALL lymphocytes PGE increased from a normal value of 25 pgrams to 270 pgrams/10(6) cells, and PGF 2 alpha increased from a normal value of 31 pgrams to 482 pgrams/10(6) cells. In CLL lymphocytes, levels of PGE and PGF2 alpha were normal or low. When normal lymphocytes were stimulated with phytohemagglutinin (PHA), the level of PGE and PGF2 alpha fluctuated, followed by corresponding changes in the level of cyclic nucleotides. In cultured ALL lymphocytes, the level of PGE remained high, while cyclic 3':5'-adenosine monophosphate (c-AMP) level was constantly low, and the initial level of PGF2 alpha fluctuated in relation to similar oscillations of cyclic 3':5'-guanosine monophosphate (c-GMP). These values were lower, although not significantly, when ALL lymphocytes were stimulated with PHA. When CLL lymphocytes were stimulated with PHA, the level of PGE remained low (20 pgrams), as did that of c-AMP. The level of PGF2 alpha, after a brief initial increase (130 pgrams), returned to and remained at a lower level (60 pgrams) while the level of c-GMP was persistently high. These results suggest: (1) prostaglandins may indirectly influence the cell cycle, possibly through modulation of cyclase activity and levels of cyclic nucleotides; and (2) some derangement of this regulatory mechanism may be present in leukemic lymphocytes.     

Observations on prostaglandins in normal and leukemic human lymphocytes

Prostaglandins E (PGE) and F₂ (PGF₂) were measured in lymphocytes of normal subjects, children with acute lymphocytic leukemia (ALL), and adults with chronic lymphocytic leukemia (CLL). In ALL lymphocytes PGE increased from a normal value of 25 pgrams to 270 pgrams/10⁶ cells, and PGF₂ increased from a normal value of 31 pgrams to 482 pgrams/10⁶ cells. In CLL lymphocytes, levels of PGE and PGF₂ were normal or low. When normal lymphocytes were stimulated with phytohemagglutinin (PHA), the level of PGE and PGF₂ fluctuated, followed by corresponding changes in the level of cyclic nucleotides. In cultured ALL lymphocytes, the level of PGE remained high, while cyclic 3′:5′-adenosine monophosphate (c-AMP) level was constantly low, and the initial high level of PGF₂ fluctuated in relation to similar oscillations of cyclic 3′:5′-guanosine monophosphate (c-GMP). These values were lower, although not significantly, when ALL lymphocytes were stimulated with PHA. When CLL lymphocytes were stimulated with PHA, the level of PGE remained low (20 pgrams), as did that of c-AMP. The level of PGF₂, after a brief initial increase (130 pgrams), returned to and remained at a lower level (60 pgrams) while the level of c-GMP was persistently high. These results suggest: (1) prostaglandins may indirectly influence the cell cycle, possibly through modulation of cyclase activity and levels of cyclic nucleotides; and (2) some derangement of this regulatory mechanism may be present in leukemic lymphocytes.     

Studies on satellite nucleoli in human normal and leukemic lymphocytes

Lymphocytes from normal and leukemic patients, and phytohemagglutinin-stimulated lymphocytes were investigated by means of immunofluorescence procedures and a silver reaction for the demonstration of proteins characteristic of nucleolus organizer regions in interphasic cells to provide basic information on the presence of satellite nucleoli in these cells. The results clearly indicated that satellite nucleoli are present in a limited but constant percentage of peripheral lymphocytes. An increased percentage of lymphocytes with satellite nucleoli was found only in leukemic patients and after silver staining. In contrast, a decreased percentage of satellite nucleoli was found 24 h after stimulation with phytohemagglutinin vitro. In leukemic patients, the discrepancy in the percentage of lymphocytes with satellite nucleoli between immunostained and silver-stained preparations may suggest that the silver reaction demonstrates the presence of an additional argyrophilic protein besides proteins B23 and C23, or altered forms of these proteins, which does not react with the specific antibodies.     

DNA polymerase activity of cultured normal and leukemic lymphocytes: Response to phytohemagglutinin

Normal lymphocytes when cultured with PHA showed increased DNA polymerase activity in 24 h. This preceded the onset of DNA synthesis. Further large increases in enzyme activity occurred in the next day or two of culture accompanying blast-cell formation and the start of mitosis.Leukemic lymphocytes from cases with low peripheral WBC counts showed the greatest increases in DNA polymerase activity and produced the largest numbers of blast-cells, while cells from cases with high WBC counts produced few or no blast-cells and failed to increase their DNA polymerase content. A broad spectrum of intermediate results were obtained in a series of 21 cases ranging in cell count from 21,000 to 265,000 per mm³.Initial studies indicate that a fluorimetric assay of cellular DNA may be a more satisfactory basis than initial cell count or protein determination for calculation of results of enzyme content of cells clumped by PHA.Increased DNA polymerase activity may be an essential preliminary to increased DNA synthesis in cells responding to PHA stimulation in cultures.     

Correlation between DNA synthesis and intracellular NAD in cultured human leukemic lymphocytes.

The NAD⁺ level in lymphocytes obtained from an individual with acute monocytic leukemia increased five-fold and then remained constant when the cells were adapted to growth in suspension culture. When the NAD⁺ level of established cells was lowered by means of a nicotinamide-poor medium or by the action of 1-methyl-1-nitrosourea, there was a concomitant decrease in the rate of DNA synthesis. These results indicate that there is a direct correlation between intracellular NAD⁺ and the synthesis of DNA in cultured leukemic lymphocytes. However, the exact nature of the relationship remains speculative.     

Ganglioside composition of normal and leukemic bovine lymphocytes

The ganglioside composition of bovine peripheral lymphocytes was shown to change sharply under lymphoid leukemia. In normal lymph, lymph nodes, spleen and blood lymphocytes the major ganglioside is N-glycolylhematoside, whereas in calf thymus lymphocytes appreciable amounts of more polar components (GM1- and GD1a-like gangliosides) were found. In leukemic lymphocytes isolated from the same tissues the hematoside content is decreased, while the amount of more polar gangliosides is increased. Possible causes of the altered ganglioside pattern in leukemic lymphocytes are discussed.     

Characteristics of the cyclic nucleotide phosphodiesterases of normal and leukemic lymphocytes

The specific activity of cylic AMP phosphodiesterase and cyclic GMP phosphodiesterase of leukemic lymphocytes was 5–10-fold greater than that of purified normal lymphocytes or homogenates of spleen, thymus or lymph nodes of normal mice. This rise was demonstrable over a wide range of substrate concentrations. Both normal and leukemic lymphocytes contained a heat-stable, calcium-dependent activator of phosphodiesterase. However, the increased activity of phosphodiesterase in leukemic lymphocytes was not due to this protein activator since (a) phophodiesterase activity from these cells was not stimulated by this activator and (b) phosphodiesterase activity of leukemic lymphocytes was not inhibited by the calcium chelator, ethyleneglycol-bis-(β-aminoethylether)-N′,N′-tetraacetic acid, suggesting that the enzyme was not already maximally activated. A comparison of several other properties of phosphodiesterase from normal and leukemic lymphocytes showed that the enzymes have similar pH optima, similar stabilitis to freezing and thawing and similar sensitivities to inhibition by the phosphodiesterase inhibitors, chlorpromazine, papaverine and isobutylmethylxanthine. However, the subcellular distribution of the phosphodiesterases was different, and the phosphodiesterase of leukemic lymphocytes was significantly more resistant to heat than that of normal lymphocytes.Although no differences were found between the phosphodiesterases of normal and leukemic lymphocytes in their sensitivities to drugs, there were marked differences in drug sensitivity between the phosphodiesterase of lymphocytes and that of other tissue. For example, concentrations of chlorpromazine which inhibited phosphodiesterase of cerebrum by 70% had no effect on phosphodiesterase activity of lymphocytes. On the other hand, the papaverine-induced inhibition of phosphodiesterase was similar in lymphocytes and cerebrum.Since an optimal concentration of cyclic nucleotides is essential to maintain normal cell growth, these results suggest that the abnormal growth characteristics of leukemic lymphocytes may be explained by their high activity of phosphodiesterase. Furthermore, the qualitative and quantitative differences between the phosphodiesterases of leukemic lymphocytes and other tissues raise the possibility of selectively inhibiting the phosphodiesterase of the leukemic lymphocytes, thereby reducing their rate of growth, without affecting other tissues.     

Glycosphingolipids of normal and leukemic human leukocytes

Summary We have reviewed the studies on neutral glycosphingolipids and gangliosides of normal and leukemia human leukocytes. In this review, we examine (a) the glycosphingolipid composition of various leukocyte populations, (b) the differences in glycosphingolipids found among subsets of these cells, (c) the possible use of these compounds as markers of differentiation, and (d) the changes in glycosphingolipid composition that occur with leukemogenesis.     

Protein concentration in normal mouse lymphocytes,thymocytes, and mouse and human leukemic cells as measured by interference microscopy

The apparent protein content of a single spherical lymphocyte or a similar cell, as well as its diameter, can be measured by a modified and stabilized AO-Baker interference microscope which is either fitted with an AO half-shade eyepiece or connected to a photomultiplier. This paper gives results for the apparent protein content and the diameter of normal mouse lymphocytes and thymocytes; these do not differ significantly from each other. It also gives values for the apparent protein concentration and the diameter of the lymphocytes of mice of the same strain which have developed leukemia or lymphoma; these values are significantly different from those of normal mice. Related data are given for the apparent protein content and for the diameter of normal human lymphocytes and the white cells found in myeloblastic leukemia and in stem cell leukemia in man; here the values differ significantly from those of the normal human lymphocyte. The rule seems to be that the abnormal white cells become more watery and larger as the disease advances.     

Enhancement by methylxanthines of sister-chromatid exchange frequency induced by cytostatics in normal and leukemic human lymphocytes

The effect of theobromine (TB) and diphylline (DP) or (1,2-dihydroxy-3-propyl)theophylline on SCE rates induced in vitro by mitomycin C (MMC), and the effect of caffeine on SCE rates induced in vitro by cytosine arabinoside (Ara-C) was studied. The combined treatments with MMC plus TB or DP showed the potentiating ability of the latter drugs. Caffeine also enhanced SCEs induced by Ara-C in cultured human lymphocytes. Caffeine and adriamycin (ADR) did not act synergistically on induction of SCEs. In a combined study, in vivo and in vitro, lymphocytes taken from 2 leukemic patients who had been given chlorambucil (CBC) or Ara-C by injection 3 h before, and then treated with caffeine in vitro, were found to have synergistically increased exchange frequencies.     

Comparative study of the ultrastructure of immune and normal phytohemagglutinin-treated lymphocytes]

Immune lymphocytes sorbed on the surface of the target cells were characterized during the period of the first three hours of combined incubation by the presence of the electron-dense matrix, abundance of mitochondria and lipids; small lymphocytes had disseminated ribosome organized into polysomes in the medium lymphocytes forming individual cysterns of the granular endoplasmic reticulum in the large lymphocytes, this indicating active protein synthesis by these cells. There were also revealed cells of plasmatic type. Cells incubated with the PHA for one hour represented a homogenous population of small lymphocytes of the same size as the clear cytoplasm containing free ribosomes and individual mitochondria. The proportion of the medium lymphocytes and the blasts increased with increase of the incubation period. These are cells with the clear cytoplasm freely disseminated polyribosomes in which no developed granular endoplasmic reticulum was sometimes revealed. The presence of two types of cells whose ultrastructure reflected their functional characteristics is discussed.     

Nucleoli of cultured human lymphocytes

Summary Nucleolar fusion and nucleolus formation occurred simultaneously, immediately after mitosis, in cultured human lymphocytes. Evidence is presented that in late telophase and post-telophase, the individual nucleolar organising site includes two components, represented in post-telophase by the nucleolus and its attached process; that fusion can be either directly between the nucleoli or via the processes; and that the latter type of fusion persists into metaphase as acrocentric associations, whilst the former is lost.