Heterogeneity of 5′-Nucleotidase Activity in Lymphocytes in Chronic Lymphocytic Leukemia

Lopes, J. D.; Zucker‐Franklin, Dorothea; Silber, Robert

doi:10.1172/jci107298

Cited by 84 publications

(17 citation statements)

References 10 publications

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“…Adenosine and inosine were obtained from Sigma Chemical Co. (St. Louis, Mo.). After informed consent had been obtained from the patients according to the provisions of the Helsinki conference, their lymphocytes were isolated from heparinized blood as previously described (10).…”

Section: Methodsmentioning

confidence: 99%

Adenosine deaminase activity in chronic lymphocytic leukemia. Relationship to B- and T-cell subpopulations.

Tung¹,

Silber²,

Quagliata³

et al. 1976

J. Clin. Invest.

131

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A B S T R A C T The level, phenotypes, and isozyme distribution of adenosine deaminase (ADA) were determined in lymphocytes from patients with chronic lymphocytic leukemia (CLL). The ADA level in lymphocytes from patients with untreated CLL was consistently lower than in lymphocytes from normal subjects. No significant differences were found in the phenotype or isozyme distribution. In untreated patients, the ADA level was inversely correlated with the lymphocyte count and the percentage of bursa-equivalent (B) cells. After therapy, a diminution in the lymphocyte count was associated with an increase of ADA activity towards normal levels. The ADA levels were slightly higher in the thymus-derived (T) than in the B lymphocytes from normal subjects. The B cells had lower activity than T cells in patients with CLL. They also had a lower activity than the B cells from normal subjects. The ADA level was 2.3-fold higher in T cells from patients with CLL than in normal T cells. The decrease in ADA levels is an anomaly that is reversible and appears to be a reflection of the proliferation of abnormal B cells in this disorder. Erythrocytes and granulocytes were separated by centrifugation through a mixture of Ficoll-Hypaque (8), while monocytes and platelets were removed on a glass wool column (9); the final preparations usually contained over 95% lymphoid cells. The criteria for selection of patients with CLL were the same as reported previously from this laboratory (10). Unless specified otherwise, the patients had never received therapy. Lymphocytes suspended in 0.05 M sodium phosphate buffer, pH 7.5, at concentrations between 5 X 10' and 2 X 108 cells/ml were disrupted by 5 cycles of .freezing and thawing followed by a 5-s exposure to ultrasound generated by a Heat Systems-Ultrasonics, Inc. cell disruptor (Plainview, N. Y.). The sonicated suspension was centrifuged at 1,100 g for 10 min and the supernatant fluid used for enzyme assays, electrophoresis, and gel-filtration studies. INTRODUCTION7The Journal of Clinical Investigation Volume 57 March 1976 -756-761 756 En.zymie assays. ADA activity was measured by the conversion of adenosine to uric acid, determined spectrophotometrically by the change in absorbance at 293 nm (11). In this assay ADA is linked to an excess of nucleoside phosphorylase and xanthine oxidase. In general, the assay, performed at 37°C in a total volume of 1 ml, contained the following: sodium phosphate buffer, pH 7.5, 50 mM; adenosine, 1.3 mM; nucleoside phosphorylase; 5 fig; xanthine oxidase, 50 ug; and lymphocyte supernatant fluid containing between 30 and 100 ug of protein. In the reaction blanks, an equal volume of 0.05 M sodium phosphate buffer, pH 7.5, was substituted for the lymphocyte supernatant fluid. Erythrocyte ADA activity was determined in a similar fashion, except for the cell disruption which was carried out by the lysis of 0.1 ml of packed erythrocytes with 4 ml of 0.01 M sodium phosphate buffer, pH 7.5, and the substitution of endogenous nucleoside phosphorylase. Protein conc...

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Section: Methodsmentioning

confidence: 99%

Adenosine deaminase activity in chronic lymphocytic leukemia. Relationship to B- and T-cell subpopulations.

Tung¹,

Silber²,

Quagliata³

et al. 1976

J. Clin. Invest.

131

View full text Add to dashboard Cite

show abstract

“…Heterogeneity for this activity has been demonstrated in chronic lymphocytic leukemia (CLL) (3,4). While the lymphocytes isolated from blood of 75% of patients with this disorder have markedly decreased or absent activity, 'Abbreviations used in this paper: CLL, chronic lymphocytic leukemia; 5'N, 5'-nucleotidase.…”

Section: Introductionmentioning

confidence: 99%

Human lymphocytes: 5'-nucleotidase-positive and -negative subpopulations.

Silber¹,

Conklyn²,

Grusky³

et al. 1975

J. Clin. Invest.

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“…Such low ecto-5'-NT activities, however, were not found consistently in the above-mentioned disease states (Lopes et al 1973;Silber et al 1975;Kanter et al 1979). In the present study ecto-5'-NT activities in both EBV-induced Ig-producing LCL and precursor B cell lines without Ig expression derived from CAG patients were not different from the enzyme activities in B LCL derived from healthy control subjects.…”

Section: Discussionmentioning

confidence: 83%

Ecto-5'-nucleotidase activities in lymphocytes and B lymphoblastoid cell lines from patients with congenital agammaglobulinemia.

Nakae

Tsuchiya

Okumura

et al. 1983

Tohoku J. Exp. Med.

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Heterogeneity of 5′-Nucleotidase Activity in Lymphocytes in Chronic Lymphocytic Leukemia

Cited by 84 publications

References 10 publications

Adenosine deaminase activity in chronic lymphocytic leukemia. Relationship to B- and T-cell subpopulations.

Adenosine deaminase activity in chronic lymphocytic leukemia. Relationship to B- and T-cell subpopulations.

Human lymphocytes: 5'-nucleotidase-positive and -negative subpopulations.

Ecto-5'-nucleotidase activities in lymphocytes and B lymphoblastoid cell lines from patients with congenital agammaglobulinemia.

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