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5'-nucleotidase activity in lymphocytes from patients affected by B-cell chronic lymphocytic leukemia.

AbstractOBJECTIVES:
The activity of membrane-bound ecto-5'-nucleotidase and soluble e-Ns and c-N-II 5'-nucleotidases was evaluated on lymphocytes from patients affected by B-cell chronic lymphocytic leukemia (B-CLL). A statistically significative decrease in ecto-5'-nucleotidase, e-Ns, and c-N-II activities was observed in peripheral blood lymphocytes and in B and T populations from affected individuals.
DESIGN AND METHODS:
For the assay of ecto-5'-nucleotidase, e-Ns, and c-N-II activity we used a radioactive procedure coupled to HPLC. Since the ecto-5'-nucleotidase is identified as CD73 antigen, we performed immunofluorescence analysis using a specific monoclonal antibody. We analyzed ecto-5'-nucleotidase mRNA by RT-PCR to ascertain the possibility of an alteration in the transcription of its gene.
RESULTS:
A decrease in ecto-5'-nucleotidase activity was correlated to reduction in ecto-5'-nucleotidase positive cells (CD73+) in leukemia patients. RT-PCR produced a fragment of the expected size and the specific mRNA was found expressed in both healthy subjects and leukemia patients.
CONCLUSIONS:
The decrease in ecto-5'-nucleotidase activity in patients with B-CLL is not due to loss of transcription of the specific mRNA. The presence of point mutations, splicing alteration, or posttranslational modifications must be investigated. If a defect at DNA or RNA level will be detected, the molecular analysis will be considered for diagnosis of B-cell chronic lymphocytic leukemia.
AuthorsF Rosi, A Tabucchi, F Carlucci, P Galieni, F Lauria, L Zanoni, R Guerranti, E Marinello, R Pagani
JournalClinical biochemistry (Clin Biochem) Vol. 31 Issue 4 Pg. 269-72 (Jun 1998) ISSN: 0009-9120 [Print] United States
PMID9646951 (Publication Type: Journal Article)
Chemical References
  • Isoenzymes
  • RNA, Messenger
  • 5'-Nucleotidase
Topics
  • 5'-Nucleotidase (blood, deficiency, genetics)
  • Aged
  • Blotting, Northern
  • Enzyme Activation
  • Fluorescent Antibody Technique
  • Humans
  • Isoenzymes (blood, deficiency)
  • Leukemia, Lymphocytic, Chronic, B-Cell (blood, enzymology)
  • Lymphocytes (enzymology)
  • Middle Aged
  • Polymerase Chain Reaction
  • RNA, Messenger (analysis)

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