Binding of Anti‐Acetaldehyde IgG Antibodies to Hepatocytes with an Acetaldehyde‐Phosphatidylethanolamine Adduct on Their Surface

Abstract

We have previously shown that antibodies raised against acetaldehyde adducts of protein cross-react with an acetaldehyde adduct of dioleoylphosphatidylethanolamine, N-ethyl-dioleoylphosphatidylethanolamine, when the latter is incorporated into hexagonal phase phospholipid micelles. In the present study we demonstrate that these same IgG antibodies cross-react with N-ethyl-dioleoylphosphatidylethanolamine when this adduct is incorporated into the surface of hepatocytes. Hapten-specific IgG antibodies were purified from the sera of rabbits sensitized to an albumin-acetaldehyde conjugate that had been reduced with sodium cyanoborohydride (N-ethyl-RSA). The N-ethyl-RSA was coupled to an Affi-Gel-10 column to affinity purify the IgG. Liposomes containing N-ethyl-dioleoylphosphatidylethanolamine were fused with isolated hepatocytes, the affinity purified primary IgG antibodies were added, then fluorescein-conjugated second antibodies were added, and antibody binding to hepatocytes was measured by flow cytometry. The fluorescence of these hepatocytes was significantly greater (p less than 0.01) than control hepatocytes prepared with (1) pre-immune primary IgG antibodies with fluorescein-conjugated second antibodies, (2) no primary antibody but with fluorescein-conjugated second antibodies, and (3) no fluorescein-conjugated second antibodies.