MONOCLONAL-ANTIBODIES AGAINST LOW-DENSITY-LIPOPROTEIN WITH VARIOUS DEGREES OF OXIDATIVE MODIFICATIONS

Oxidative processes leading to the generation of oxidized low density lipoprotein (oxLDL) particles have been suggested to be an important f actor in the pathogenesis of atherosclerosis. After initiation of the oxidative process, LDL undergoes a progressive protein and lipid fragm entation, To understand this process and the role of oxLDL in various diseases of inflammatory origin, we have generated mouse monoclonal an tibodies against copper-oxidized human LDL. Mice were immunized intras plenically and after one intravenous boost the spleen cells were fused with the Sp2/0 hybridoma fusion partner. The hybridoma clones obtaine d after selection and cloning were analyzed for reactivity against oxL DL with various degrees of copper-mediated oxidative modifications. Th ree hybridoma clones were purified and further characterized. The foll owing observations were made: I)the intrasplenic route of immunization , avoiding the use of mycobacterial adjuvants, yielded a high frequenc y of positive clones; 2) the individual hybridomas reacted against LDL with various degrees of oxidative modifications; 3) the monoclonal an tibodies could be used in ELISA and to detect oxLDL in immunohistochem ical tissue staining, and 4) the monoclonal antibodies also detected o xLDL from hamsters and rabbits, We conclude that these monoclonal anti bodies could be useful to further investigate the role of oxLDL in inf lammation and in the immune response.