Approach to reduction of blood atherogenicity

Abstract

We have earlier found that blood sera of patients with coronary heart disease (CHD) increase lipid levels in cells cultured from subendothelial intima of human aorta. We have also revealed that the ability of blood sera to raise intracellular cholesterol; that is, their atherogenicity is caused by at least modified low density lipoprotein (LDL) circulating in the blood of patients and autoantibodies to modified LDL. In the present work we have demonstrated significant impact of nonlipid factor(s) to blood atherogenicity. We have developed an approach to removal of nonlipid atherogenicity factor(s) from blood serum based on the use of immobilized LDL. This approach was used for extracorporeal perfusion of patient's blood through the column with immobilized LDL. Pilot clinical study confirmed the efficacy of this approach for prevention of coronary atherosclerosis progression.

Figure 1

Elimination of serum atherogenicity with LDL-agarose column. Five milliliters of the serum was passed through the LDL-sepharose column at a flow rate of 1 mL/min for 30 min. The sorbent was then eluted with 2 mL glycine buffer (pH 2.7), and the eluate was dialyzed against a 2,000-fold excessive volume of medium 199 for 24 hours at 4°C. The cells were cultured in the presence of the initial or treated serum and with the proper volume of the dialyzed eluate.

Figure 2

Monitoring of atherogenicity. The patient's plasma was subjected to 2-hour extracorporeal perfusion through a column with 200 mL of the sorbent; the flow rate was 30 mL/min. The total plasma volume of 2-3 liters was perfused during the procedure. The serum atherogenicity after 3 procedures was assessed daily ((a), patient 3) and once or twice a week afterwards ((b), patient 1). Ordinate (atherogenicity), % of cholesterol accumulation in the cells cultured in the presence of the serum from the CHD patient.