Redox buffering ability of lymphoid cells evaluated by the oxidation of 2'7'dichlorofluorescin

Gennaro Melino, Isabella Savini, Pietro Guerrieri, Alessandro Finazzi-Agrò

Research output: Contribution to journalArticle

6 Citations (Scopus)

Abstract

The redox buffering activity of several lymphoid cells against endogenous and exogenous H2O2 has been evaluated using 2'7'dichlorofluorescin diacetate (DCFH2-DA). The mechanism of 2'7'dichlorofluorescin (DCFH2) oxidation has also been investigated. It was found that while the oxidation by external H2O2 is completely inhibited by azide or cyanide, the oxidation by endogenous species is still present, even under anaerobic conditions. The data herein reported indicate that autoxidation and peroxidation of DCFH2 are distinct reactions. Hence only by addition of increasing concentrations of exogenous hydrogen peroxide, the fluorescence of DCF can be used to evaluate the cellular ability of scavenging H202. By this method we have found that the erythroleukaemia cell line K56 and promyelocytic line HL-60 show a faster rate of DCFH2 oxidation than peripheral blood leukocytes (PBL), mature T-cells (MOLT-3 and MOLT-4) and B-cells (DAUDI). Using this method the balance between antioxidant enzymes activity and the redox state of the cell can be easily assessed by fluorescence both in single cells and in cell populations.

Original languageEnglish
Pages (from-to)213-221
Number of pages9
JournalFree Radical Research
Volume11
Issue number4-5
DOIs
Publication statusPublished - 1990

Fingerprint

Oxidation-Reduction
Cells
Lymphocytes
Oxidation
Fluorescence
Leukemia, Erythroblastic, Acute
Azides
Cyanides
Hydrogen Peroxide
T-cells
Scavenging
Enzyme activity
Leukocytes
B-Lymphocytes
Antioxidants
T-Lymphocytes
Cell Line
Blood
Enzymes
Population

Keywords

  • 2'7'dichlorofluorescin diacetate
  • Catalase
  • Lymphocytes
  • Oxygen radicals
  • Peroxidase

ASJC Scopus subject areas

  • Biochemistry

Cite this

Redox buffering ability of lymphoid cells evaluated by the oxidation of 2'7'dichlorofluorescin. / Melino, Gennaro; Savini, Isabella; Guerrieri, Pietro; Finazzi-Agrò, Alessandro.

In: Free Radical Research, Vol. 11, No. 4-5, 1990, p. 213-221.

Research output: Contribution to journalArticle

Melino, Gennaro ; Savini, Isabella ; Guerrieri, Pietro ; Finazzi-Agrò, Alessandro. / Redox buffering ability of lymphoid cells evaluated by the oxidation of 2'7'dichlorofluorescin. In: Free Radical Research. 1990 ; Vol. 11, No. 4-5. pp. 213-221.
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