Biological Trace Element Research

, Volume 47, Issue 1–3, pp 17–27 | Cite as

Immunological approach to investigating membrane cell damages induced by lipoperoxidative stress

Application to far UV-irradiated erythrocytes
  • E. Petit
  • D. Divoux
  • Y. Chancerelle
  • J. F. Kergonou
  • A. Nouvelot
Part I Free Radical Stress: Interactions with Trace Elements or Vitamins

Abstract

Oxygen-reactive species are being described as agents responsible for cell degeneration mechanisms resulting from membrane, enzyme, and nuclear alterations. Lipid peroxidation on its own is considered to be one of the consequences of the free radicals attack, and among the different reactive aldehydes that can be formed from the decomposition of lipid peroxides, the most extensively assayed have been malondialdehyde (MDA). However, the different techniques currently used for MDA assay (HPLC, GLC) are barely sensitive enough to follow its production at the cellular level.

In order to develop an immunofluorescent technique able to detect cellular damages provoked by lipoperoxidation, polyclonal antibodies against lysozyme modified by MDA treatment have been raised in rabbits. We show that this immunserum recognizes specifically all the MDA-treated proteins tested, but not the intact proteins or the proteins treated by other aldehydes.

Moreover, we demonstrate using an ELISA technique that the amount of immunoreactive proteins in MDA-treated membrane erythrocytes is proportional to the concentration of MDA applied, suggesting that this assay may represent a quantitative method of determination of lipoperoxidative alterations. In addition, when coupled to an indirect fluorophore antibody (FITC), the immunserum allows a precise location of these modified proteins within the membranes of erythrocytes in which lipid peroxidation was initiated by far UV irradiation.

In summary, the interest of this work is to provide an immunological probe that can precociously detect membrane damages induced by MDA, regardless of the cell type and pro-oxidant (physiological or pathological) conditions.

Keywords

Lipid Peroxidation Lysozyme Biological Trace Element Research Erythrocyte Ghost Reactive Aldehyde 
These keywords were added by machine and not by the authors. This process is experimental and the keywords may be updated as the learning algorithm improves.

Abbreviations

ELISA

enzyme-linked immunosorbent assay

GLC

gas-liquid chromatography

HPLC

high-performance liquid chromatography

LDL

low-density lipoproteins

MDA

malondialdehyde

TBA

thiobarbituric acid

TLC

thin-layer chromatography

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Copyright information

© Humana Press Inc. 1995

Authors and Affiliations

  • E. Petit
    • 1
  • D. Divoux
    • 1
  • Y. Chancerelle
    • 2
  • J. F. Kergonou
    • 2
  • A. Nouvelot
    • 1
  1. 1.Laboratoire de NeurosciencesURA 1829-CNRSCaen, Cedex
  2. 2.Centre de Recherches du Service de Santé des ArméesLa Tronche, Cedex

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