Free fatty acids released from phospholipids are the major heat-stable hemolytic factor of Entamoeba histolytica trophozoites

Abstract

The major hemolytic activity of Entamoeba histolytica trophozoites is located in a vesicular fraction called P30 and known to be due to heat-labile and heat-stable hemolytic components whose effect increases up to 100 times during preincubation at 36 degrees C. The heat-stable hemolytic activity (HSHA) was found in the chloroform-methanol extract of preincubated P30, whose partition with 2 M KCl yielded a lipid fraction, an interphase, and an aqueous phase. HSHA was detected only in the lipid fraction, where it amounted to 59% of the chloroform-methanol extract activity and increased 50% when supplemented with the interphase material; it was accounted for by the free fatty acids, whose potency increased 33% with the interphase material, and was blocked by delipidated bovine serum albumin. A parallel increase in free fatty acids and lysophospholipids and a corresponding decrease in phospholipids were observed during P30 preincubation. Most of the phospholipase activity of trophozoite homogenates was also found in P30. Therefore, most of the HSHA generated during preincubation was due to free fatty acids released from phospholipids by a P30 phospholipase that may contribute significantly to E. histolytica cytopathogenicity and virulence.