Arteriosclerosis, Thrombosis, and Vascular Biology. 1995;15:982-989
© 1995 American Heart Association, Inc.
Articles |
Immunologic Detection and Measurement of Hypochlorite-Modified LDL With Specific Monoclonal Antibodies
From Karl-Franzens University, Institutes of Medical Biochemistry (E.M., W.S.) and Biochemistry (H.E., G.W.), Graz, Austria, and the Heart Research Institute, Biochemistry Group, Camperdown, Australia (L.H., R.S.).
Abstract Oxidation of LDL is thought to contribute to the
early stages of atherogenesis. Because myeloperoxidase is present
in atherosclerotic lesions and can produce the strong oxidant
hypochlorous acid (HOCl), which converts LDL into its high-uptake
atherogenic form in vitro, we raised polyclonal and monoclonal
antibodies (MoAbs) against HOCl-modified LDL (HOCl-LDL).
Characterization of the polyclonal anti-human HOCl-LDL Abs showed that
they cross-reacted strongly with 4-hydroxynonenal–, malondialdehyde-,
and Cu2+-oxidized LDL. Similarly, polyclonal and some
monoclonal Abs against aldehyde- and Cu2+-modified LDL
cross-reacted with HOCl-LDL. In contrast to the polyclonal Abs, two
selected hybridoma cell line supernatants containing MoAbs raised
against HOCl-LDL (MoAb-A and MoAb-B) did not cross-react with either
native LDL or aldehyde- or Cu2+-modified LDL. MoAb-A (clone
1B10A11, subtype IgG1
) recognized an epitope that appeared to be
specific for HOCl-LDL and depended on the tertiary structure of the
(lipo)protein, as judged by a lack of cross-reactivity with
HOCl-modified human and bovine serum albumin and a loss of reactivity
associated with lipoprotein denaturation. MoAb-B (clone 2D10G9, subtype
IgG2b), on the other hand, gave identical titration curves with
HOCl-LDL and HOCl-modified albumins, suggesting that this antibody
recognized epitopes that are commonly generated on proteins that have
been oxidized with HOCl. Thus, MoAb-A and MoAb-B may be useful tools
for the investigation of a possible role for HOCl-mediated damage to
(lipo)proteins in atherosclerosis and other inflammatory diseases.
Key Words: myeloperoxidase • lipid peroxidation • atherosclerosis • oxidized lipoproteins
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