Isolated high density lipoprotein (HDL) from the two living fish eye disease patients have been incubated in vitro with autologous lipoprotein depleted plasma or with lipoprotein depleted plasma from domestic pig (Sus domesticus), with and without the presence of LCAT inhibitor for 24 hours at 0 and 37 degrees C. The lecithin:cholesterol acyltransferase (LCAT) activity in lipoprotein depleted pig plasma increased the abnormally low cholesteryl ester content of the fish eye disease HDL particles from about 20 to 100% and increased their exceptionally small mean particle size, probably by particle fusion, to a range which is representative of normal HDL3. Both esterification and particle enlargement were totally blocked by the LCAT inhibitor. Incubation of concentrated fish eye disease HDL with autologous lipoprotein depleted plasma for 24 hours at 37 degrees C resulted in a small increase in its cholesteryl ester percentage to 37%, without affecting the apparent HDL particle size. This finding confirms a deficiency of HDL lecithin:cholesterol acyltransferase activity (alpha-LCAT) in fish eye disease. The observed normalization of both HDL cholesteryl ester percentage and particle size by lipoprotein depleted pig plasma which contains virtually no cholesteryl ester transfer activity indicates that the latter is not a requisite for esterification of the free cholesterol of fish eye disease HDL.