Restoration of in-Vitro Lymphocyte Responses With Exogenous Adenosine Deaminase in a Patient With Severe Combined Immunodeficiency

Polmar, StephenH.; Wetzler, EricaM.; Stern, R C; Hirschhorn, Rochelle

doi:10.1016/s0140-6736(75)90726-6

Cited by 122 publications

(29 citation statements)

References 14 publications

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“…It is, therefore, tempting to speculate that the functionally abnormal B cell in CLL patients may be related to the lower levels of ADA, as appears to be the case for the lymphocytes from combined immunodeficiency patients. Experiments are underway to test the hypothesis that addition of exogenous ADA will enhance the in vitro response of CLL lymphocytes, as it does for lymphocytes from combined immunodeficiency patients (26).…”

Section: Resultsmentioning

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: Resultsmentioning

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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“…Unfortunately, over half the children do not have a suitable sibling donor. Therefore, based upon the results of in vitro experiments (16), children have been treated with multiple partial exchange transfusions with normal frozen, irradiated erythrocytes (4). These erythrocytes contain both the missing enzyme and nucleoside transport sites for the accumulated substrates (7).…”

Section: Introductionmentioning

confidence: 99%

Plasma deoxyadenosine, adenosine, and erythrocyte deoxyATP are elevated at birth in an adenosine deaminase-deficient child.

Hirschhorn¹,

Roegner²,

Rubinstein³

et al. 1980

J. Clin. Invest.

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“…Current therapies for Ado-deaminase deficiency include erythrocyte transfusion [28], weekly or bi-weekly injection with expensive poly(ethy1ene glycol)-modified Ado deaminase [29], bone-marrow transplantation [30] and, most recently, gene therapy [31]. Nucleoside chemotherapy with dCyd has also been attempted, but with limited success [32].…”

Section: Discussionmentioning

confidence: 99%

The bis(adenosin‐N⁶‐yl) alkanes, a family of potential dinucleoside polyphosphate analogue precursors

Chen

McLennan

1993

European Journal of Biochemistry

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The potential diadenosine polyphosphate analogue precursor, bis(adenosin-W-yl)dodecane (A[CH,],2A) (Chen, H. & McLennan, A. G. (1993) Eul: J. Biochem. 213, 935-944.) is equally toxic to both wild-type and adenosine-kinase-deficient BHK cells at concentrations up to 100 pM ; at higher concentrations, wild-type cells are more sensitive, as are cells over-expressing adenosine kinase. Thus both the nucleoside and its nucleotide products are toxic. In contrast to adenosine toxicity, the toxicity of A[CH,],,A to S-49 T-lymphoma cells could not be reversed by uridine or by L-homocysteine thiolactone. A [CH,] ,,A and all its shorter chain bis(adenosin-W-yl)alkane homologues could relieve the toxicity of low adenosine concentrations (< 20 pM) to $49 cells, mainly through inhibition of adenosine kinase, while relief of the toxicity of high adenosine concentrations (>20 pM) required the longer chain homologues. A [CH,],,A at 10 pM completely eliminated adenosine toxicity. Deoxyadenosine toxicity could also be relieved, but only that due to low concentrations (< 4 pM). A[CH,] ,,A had only a slight stimulatory effect on S-adenosylhomocysteinehydrolase activity. The bis(adenosin-W-y1)alkancs (A[CH2].A) comprise a series of compounds in which two adenosine residues are linked through their W-amino groups by alkyl chains containing up to 14 methylene units [l, 21. They were synthesized with the intention that they would behave as cell-permeable precursors to analogues of nucleoside(5')polyphospho(5')nucleosides (NpnN), such as adenosine(5')tetraphospho(5')adenosine (ApJA), and therefore be useful as tools for determining the functions of these unusual nucleotides [3].We have previously described some of the toxicological, receptor binding and metabolic properties of these compounds [4]. They are moderately toxic towards a variety of mammalian cell lines; they bind preferentially to A, Ado receptors, although they are also weak agonists at A, receptors, causing an increase in intracellular CAMP ; they are phosphorylated by Ado kinase and adenylate kinase in vitro and in vivo to yield a number of products which have the potential to act as Ap,A analogues.In this study, we show that the biochemical basis for their cytotoxicity is quite distinct from that of Ado and that they are able to relieve completely Ado toxicity and partially relieve dAdo toxicity towards lymphoid cells, a finding that may be relevant to the treatment of hereditary Ado-deaminase deficiency. Some of these results have previously been reported in brief [5]. MATERIALS AND METHODS MaterialsCell lines and reagents were as previously described [4]. In addition, the BHK ara-Sl0d Ado-kinase mutant [6] was kindly provided by V.-L. Chan, University of Toronto and Bmix AK' Rous-sarcoma-virus-transformed rat embryo fibrohlnsts (A&J clrarninase deficicnt) and B=mix A K ~ cclL (additionally Ado-kinase deficient) 171 were the generous gift of J. Drach, University of Michigan. L-Homocysteine thiolactone (Hcy thiolactone) was from Sigma. Neplanocin A was kindly provided b...

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Restoration of in-Vitro Lymphocyte Responses With Exogenous Adenosine Deaminase in a Patient With Severe Combined Immunodeficiency

Cited by 122 publications

References 14 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.

Plasma deoxyadenosine, adenosine, and erythrocyte deoxyATP are elevated at birth in an adenosine deaminase-deficient child.

The bis(adenosin‐N⁶‐yl) alkanes, a family of potential dinucleoside polyphosphate analogue precursors

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Restoration of in-Vitro Lymphocyte Responses With Exogenous Adenosine Deaminase in a Patient With Severe Combined Immunodeficiency

Cited by 122 publications

References 14 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.

Plasma deoxyadenosine, adenosine, and erythrocyte deoxyATP are elevated at birth in an adenosine deaminase-deficient child.

The bis(adenosin‐N6‐yl) alkanes, a family of potential dinucleoside polyphosphate analogue precursors

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The bis(adenosin‐N⁶‐yl) alkanes, a family of potential dinucleoside polyphosphate analogue precursors